Method, system and equipment for evaluating treatment effect of ST14 accelerant on lung cancer based on TKT glycosylation modification level

Abstract

The invention discloses a method, a system and equipment for evaluating a lung cancer treatment effect of an ST14 accelerant based on a TKT glycosylation modification level. The invention firstly discovers that the ST14 accelerant plays a role in treating the lung cancer by inhibiting the TKT O-GlcNAc glycosylation modification level, based on this, the invention provides a method, a system and equipment for evaluating the lung cancer treatment effect of the ST14 accelerant based on the O-GlcNAc-TKT expression level for the field, and the method, the system and the equipment have wide application prospects in the aspect of lung cancer gene drug treatment effect evaluation.

Description

Technical Field

[0001] The present invention belongs to the field of bioinformatics. Specifically, the present invention relates to a method, system and device for evaluating the therapeutic effect of ST14 promoter on lung cancer based on TKT glycosylation modification level. Background Art

[0002] Lung cancer is the malignant tumor with the highest morbidity and mortality rate worldwide. Among the top ten malignant tumors in my country, lung cancer ranks first in mortality and is the leading cause of cancer death in men. Histologically, 80% to 85% of lung cancers are non-small cell lung cancer (NSCLC). Since most NSCLC patients are diagnosed in the locally advanced stage, they have lost the opportunity for surgical cure. Therefore, medical treatments, including chemotherapy, molecular targeted therapy, immunotherapy, and gene therapy, play a vital role in the treatment of advanced NSCLC.

[0003] The ST14 gene plays a crucial role in normal physiological processes, and abnormal expression levels are associated with the development and progression of certain cancers. Existing studies have reported that ST14 acts as a tumor suppressor gene in non-small cell lung cancer. Therefore, to evaluate the efficacy of ST14 gene therapy for lung cancer, it is crucial to identify biomarkers that can guide ST14 treatment in non-small cell lung cancer. Summary of the Invention

[0004] In order to make up for the deficiencies of the prior art, the purpose of the present invention is to screen a biomarker for evaluating the therapeutic effect of ST14 promoter on lung cancer.

[0005] In order to achieve the above purpose, the present invention adopts the following technical means: A first aspect of the present invention provides a method for evaluating the therapeutic effect of an ST14 promoter on lung cancer.

[0006] Furthermore, the method is performed by a computer and comprises the following steps: Obtain data: Obtain O-GlcNAc-TKT expression data in lung cancer patient samples after ST14 promoter treatment; Processing data: inputting the O-GlcNAc-TKT expression data into a constructed evaluation model, wherein the evaluation model predicts the therapeutic effect of the ST14 promoter on lung cancer patients based on the O-GlcNAc-TKT expression data; Output the results.

[0007] In the present invention, the O-GlcNAc-TKT refers to the O-GlcNAc glycosylation modification level of TKT protein.

[0008] Furthermore, the steps for constructing the evaluation model are as follows: Obtaining O-GlcNAc-TKT expression data from untreated lung cancer patients and lung cancer patients treated with an ST14 promoter; inputting the O-GlcNAc-TKT expression data into a machine learning algorithm to construct an evaluation model.

[0009] Furthermore, the machine learning algorithm includes algorithm models developed using various development tools.

[0010] Furthermore, the development tools include but are not limited to TensorFlow, Scikit Learn, PyTorch, OpenNN, RapidMiner, Azure Machine Learning, Apache Mahout, Shogun, KNIME, Vertex AI, H2Oai, Anaconda, Keras, Tableau, Fast.ai, Catalyst, Amazon ML, MLJAR, and Spell.

[0011] Furthermore, the algorithm model includes but is not limited to linear regression model, logistic regression model, Lasso regression model, Ridge regression model, linear discriminant analysis model, nearest neighbor model, decision tree model, perceptron model, neural network model, support vector machine model, naive Bayes model, AdaBoost model, GBDT model, XGBoost model, LightGBM model, CatBoost model, and random forest model.

[0012] Furthermore, the evaluation model obtains results through the following criteria: When the expression level of the O-GlcNAc-TKT is lower than the threshold, a classification result is obtained that the ST14 promoter is effective in treating lung cancer patients; when the expression level of the O-GlcNAc-TKT is higher than the threshold, a classification result is obtained that the ST14 promoter is ineffective in treating lung cancer patients.

[0013] A second aspect of the present invention provides a system for evaluating the therapeutic effect of an ST14 promoter on lung cancer.

[0014] Furthermore, the system comprises: Data acquisition unit: used to obtain the expression data of O-GlcNAc-TKT in lung cancer patient samples after ST14 promoter treatment; A data classification unit is used to classify and predict the data obtained by the data acquisition unit using the evaluation model obtained by the construction method described in the first aspect of the present invention, to obtain a classification result of whether the ST14 promoter is effective in treating lung cancer patients; Result output unit: used to output classification results.

[0015] A third aspect of the present invention provides a computer device.

[0016] Furthermore, the computer device includes a memory and a processor, the memory is used to store program instructions; the processor is used to call the program instructions, and when the program instructions are executed, the method for evaluating the therapeutic effect of ST14 promoter on lung cancer described in the first aspect of the present invention is implemented.

[0017] A fourth aspect of the present invention provides a computer-readable storage medium.

[0018] Furthermore, the computer-readable storage medium stores a computer program, and when the computer program is executed by the processor, the method for evaluating the therapeutic effect of the ST14 promoter on lung cancer described in the first aspect of the present invention is implemented.

[0019] A fifth aspect of the present invention provides a method for regulating the expression of O-GlcNAc-TKT in lung cancer cells.

[0020] Furthermore, the method includes: in an in vitro environment, ST14 negatively regulates the expression level of O-GlcNAc-TKT in lung cancer cells; by regulating the expression level of ST14, the regulation of O-GlcNAc-TKT expression in lung cancer cells can be achieved.

[0021] Furthermore, when ST14 expression was upregulated, the expression level of O-GlcNAc-TKT decreased; when ST14 expression was inhibited, the expression level of O-GlcNAc-TKT increased.

[0022] Furthermore, the upregulation of ST14 expression is achieved through gene overexpression technology.

[0023] Furthermore, the gene overexpression vector includes a eukaryotic expression vector and a viral vector.

[0024] A sixth aspect of the present invention provides use of an ST14 promoter in the preparation of a drug for treating lung cancer.

[0025] Furthermore, the ST14 promoter exerts its effect by inhibiting the expression level of O-GlcNAc-TKT.

[0026] Advantages and beneficial effects of the present invention: The present invention discovers for the first time that the ST14 promoter plays a role in treating lung cancer by inhibiting the level of TKT O-GlcNAc glycosylation modification. Based on this, the present invention provides a method, system and equipment for evaluating the therapeutic effect of the ST14 promoter on lung cancer based on the O-GlcNAc-TKT expression level in the field, which has broad application prospects in the evaluation of the therapeutic effect of gene drugs for lung cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 A flow chart of a method for evaluating the therapeutic effect of an ST14 promoter on lung cancer provided by an embodiment of the present invention; Figure 2 A schematic diagram of a system for evaluating the therapeutic effect of ST14 promoter on lung cancer provided by an embodiment of the present invention; Figure 3 A schematic diagram of the structure of the computer device provided by the present invention; Figure 4 Figure 1 is the result of ST14 promoter inhibiting lung cancer cell proliferation; A is Western blot verification of HA-ST14 expression in A549 and H1299 overexpression stable transfectants; B is the CCK8 experiment result showing that overexpression of ST14 in A549 and H1299 decreased cell proliferation ability; C is the clone formation experiment result showing that overexpression of ST14 in A549 and H1299 decreased cell proliferation ability; D is Western blot Verification of ST14 expression in A549 and H1299 knockdown stable transfectants; E is the CCK8 assay result showing that knocking down ST14 in A549 and H1299 enhanced cell proliferation; F is the clone formation assay result showing that knocking down ST14 in A549 and H1299 enhanced cell proliferation; G1 is the nude mouse subcutaneous tumor model constructed using ST14-overexpressing A549 stable transfectants, and the results showed that overexpression of ST14 significantly reduced tumor weight (H) and volume (I) compared with the control group cells; J is the Ki67 immunohistochemical analysis of subcutaneous tumors, showing that overexpression of ST14 reduced tumor cell proliferation energy; Figure 5 ST14 inhibits lung cancer cell proliferation by inhibiting O-GlcNAc glycosylation modification; A is a Western blot test showing that compared with adjacent adjacent tissues, ST14 expression is downregulated and protein glycosylation levels are increased in lung cancer tissues; B is a CCK8 assay showing that treatment of ST14-overexpressing A549 and H1299 stably transfected cell lines with TG as the glycosyltransferase inhibitor restores cell proliferation; C is a clone formation experiment further confirming that the inhibitory effect of ST14 on the proliferation of A549 and H1299 can be reversed by glycosyltransferase inhibitors; D is a CCK8 assay showing that treatment of ST14-knockdown A549 and H1299 stably transfected cell lines with OGT-IN as the glycosylation inhibitor reduces cell proliferation; E is a clone formation experiment further confirming that the promoting effect of ST14 knockdown on lung cancer cell proliferation can be reversed by OGT-IN; Figure 6ST14 inhibits TKT O-GlcNAc glycosylation by inhibiting the binding of OGT to TKT; A shows that ST14 binding mass spectrometry and O-GlcNAc glycosylation mass spectrometry were performed on A549, respectively, and 319 proteins bound to ST14 were found to be O-GlcNAc glycosylated; BC shows that further Go and KEGG signaling pathway enrichment analysis was performed on the 319 proteins that were O-GlcNAc glycosylated, and the results showed significant changes in glycolysis and pentose phosphate pathways; D shows that ST14 was overexpressed in A549 and H1299, and TKT expression did not change significantly, but O-GlcNAc-TKT expression was significantly downregulated; E shows that ST14 was knocked down, and TKT expression did not change significantly, but O-GlcNAc-TKT expression was increased; F shows that protein immunoprecipitation experiments confirmed that OGT and TKT can bind to each other; G shows that the glycosyltransferase inhibitor TG can promote TKT O-GlcNAc Glycosylation modification; H: knockdown of ST14 in A549, which enhanced the binding of OGT and TKT; I: overexpression of ST14 in A549, which weakened the binding of OGT and TKT; Figure 7 Figure 3: TKT regulation of lung cancer cell proliferation and glycolysis pathway depends on O-GlcNAc glycosylation modification; A is the SWGA experimental results showing that TKT 30, 31 threonine and 345 serine affect its O-GlcNAc glycosylation modification; B is TKT with triple mutants of threonine 30, 31 and 345 serine cannot undergo O-GlcNAc glycosylation modification; C is Western blot verification of Flag-TKT WT and Flag-TKT mutant expression in cell lines; D is CCK8 results showing that compared with wild-type TKT, TKT glycosylation site mutants have a significantly weakened promoting effect on cell proliferation; E is a clone formation experiment further confirming that TKT's promoting effect on lung cancer cell proliferation depends on its O-GlcNAc glycosylation modification; F is TKT's promoting effect on glucose consumption in lung cancer cell lines depends on O-GlcNAc Glycosylation modification; G represents the effect of TKT on promoting lactate production in lung cancer cell lines, which is dependent on O-GlcNAc glycosylation modification; Figure 8The regulatory effect of ST14 on the proliferation and glycolysis pathway of lung cancer cell lines depends on TKT; A is the CCK8 result showing that overexpression of TKT in the case of overexpression of ST14 can significantly reverse the inhibitory effect of ST14 on the proliferation of lung cancer cell lines; B is the clone formation experiment result showing that overexpression of TKT in the case of overexpression of ST14 can significantly reverse the inhibitory effect of ST14 on the proliferation of lung cancer cell lines; C is the case of overexpression of ST14, overexpression of TKT, cellular glucose consumption increased significantly; D is the case of overexpression of ST14 Overexpression of TKT significantly increased cellular lactate production; E is the CCK8 result showing that knocking down TKT in the case of knocking down ST14 can significantly reverse the promoting effect of knocking down ST14 on the proliferation of lung cancer cell lines; F is the result of the clone formation experiment showing that knocking down TKT in the case of knocking down ST14 can significantly reverse the promoting effect of knocking down ST14 on the proliferation of lung cancer cell lines; G is the case of knocking down ST14, knocking down TKT, significantly reduced cellular glucose consumption; H is the case of knocking down ST14, knocking down TKT, significantly reduced cellular lactate production; Figure 9 The regulation of lung cancer cell proliferation and glycolysis by ST14 depends on TKT O-GlcNAc glycosylation modification; A is the CCK8 result showing that under the condition of double knockdown of ST14 and TKT, overexpression of TKT can reverse the promoting effect of ST14 knockdown on the proliferation of lung cancer cell lines, while overexpression of TKT glycosylation mutants has no obvious reversal effect; B is the clone formation experiment result showing that under the condition of double knockdown of ST14 and TKT, overexpression of TKT can reverse the promoting effect of ST14 knockdown on the proliferation of lung cancer cell lines, while overexpression of TKT glycosylation mutants has no obvious reversal effect; C is under the condition of double knockdown of ST14 and TKT, overexpression of TKT promotes cellular glucose consumption and lactate production, while overexpression of TKT glycosylation mutants has no obvious promoting effect. DETAILED DESCRIPTION

[0028] In order to make up for the shortcomings of the existing technology, the overall idea of ​​the technical solution provided in this application is as follows: by overexpressing ST14 in lung cancer cells A549 and H1299, it was found that ST14 plays an important role in the treatment of lung cancer; then, through analysis, it was found that in lung cancer cells, ST14 negatively regulates the level of TKT O-GlcNAc glycosylation; based on this, we propose a method, system and equipment for evaluating the therapeutic effect of ST14 promoter on lung cancer based on O-GlcNAc-TKT.

[0029] In order to enable those skilled in the art to better understand the solutions of the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the accompanying drawings in the embodiments of the present invention.

[0030] In some of the processes described in the specification and claims of the present invention and the accompanying drawings, multiple operations are included in a specific order. However, it should be understood that these operations may not be performed in the order in which they are presented herein or may be performed in parallel. Operation numbers such as 101, 102, etc. are merely used to distinguish between different operations and do not represent any order of execution. In addition, these processes may include more or fewer operations, and these operations may be performed sequentially or in parallel.

[0031] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments in the present invention, all other embodiments obtained by those skilled in the art without making creative work are within the scope of protection of the present invention. The experimental methods in the following examples where specific conditions are not specified are usually tested under conventional conditions or under the conditions recommended by the manufacturer.

[0032] Figure 1 A flow chart of a method for evaluating the therapeutic effect of an ST14 promoter on lung cancer provided by an embodiment of the present invention is provided. Specifically, the method comprises the following steps: 101: Obtain data, obtain the expression data of O-GlcNAc-TKT in lung cancer patient samples after ST14 promoter treatment; In certain embodiments, the patient can be human or non-human, and can include, for example, an animal strain or species used as a "model system" for research purposes. Likewise, the patient can include an adult or adolescent (e.g., child). In addition, the patient can refer to any living organism that can benefit from the administration of the ST14 promoter herein, preferably mammals (e.g., humans or non-humans). The example of mammals includes, but is not limited to, any member of the mammalian class: humans, non-human primates (e.g., chimpanzees) and other apes and monkeys; livestock, such as cattle, horses, sheep, goats, pigs; livestock, such as rabbits, dogs and cats; experimental animals include rodents, such as rats, mice and guinea pigs, etc. The example of non-mammals includes, but is not limited to, birds, fish, etc.

[0033] As used herein, the term "sample" refers to a composition obtained from or derived from a patient / subject that contains cells and / or other molecular entities to be characterized and / or identified based on, for example, physical, biochemical, chemical, and / or physiological characteristics. For example, a sample refers to any sample derived from a patient / subject that is expected or known to contain cells and / or molecular entities to be characterized. Samples include, but are not limited to, tissue samples, primary or cultured cells or cell lines, cell cultures, cell supernatants, cell lysates, platelets, serum, plasma, vitreous humor, lymph fluid, synovial fluid, follicular fluid, semen, pancreatic juice, amniotic fluid, breast milk, whole blood, blood-derived cells, urine, cerebrospinal fluid, saliva, sputum, tears, sweat, mucus, tissue culture fluid, tissue extracts, homogenized tissues, cell extracts, and combinations thereof. In a specific embodiment of the present invention, the sample comprises human lung cancer cell lines A549 and H1299. A549 and H1299 cells were routinely cultured in a cell culture incubator (37°C, 5% CO2) in DMEM high-glucose medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin solution (hereinafter referred to as double antibody).

[0034] In some embodiments, the ST14 promoter refers to any substance that can increase the activity of the ST14 protein, improve the stability of the ST14 gene or protein, upregulate the expression of the ST14 protein, increase the effective action time of the ST14 protein, or promote the transcription and translation of the ST14 gene, and these substances can be used in the present invention. In a specific embodiment of the present invention, the promoter is a viral vector that promotes overexpression of ST14.

[0035] In the present invention, the O-GlcNAc-TKT refers to the glycosylation modification level of O-GlcNAc of TKT protein. In some embodiments, the expression level data of O-GlcNAc-TKT can be detected by using methods well known in the art. For example, the expression level data of O-GlcNAc-TKT can be obtained by measuring the amount of RNA, mRNA or any other RNA species using methods well known in the art to obtain expression at the nucleic acid level, including digital PCR and real-time (RT) quantitative or semi-quantitative PCR, fluorescence activated cell sorting (FACS) and in situ hybridization.

[0036] In other embodiments, the expression level data of O-GlcNAc-TKT can also be obtained by measuring the expression level at the protein level, including mass spectrometry-based quantitative proteomics technology, immunoassay, Western blotting, spectrophotometry, enzyme assay, ultraviolet assay, kinetic assay, electrochemical assay, colorimetric assay, turbidimetric assay, atomic absorption assay, flow cytometry, mass spectrometry flow cytometry, or any combination thereof. In a specific embodiment of the present invention, Western blotting is used to detect the expression level of O-GlcNAc-TKT.

[0037] In one embodiment of the present invention, we demonstrated that the ST14 gene has the effect of inhibiting lung cancer cell proliferation and treating lung cancer. First, we overexpressed ST14 in lung cancer cell lines A549 and H1299. Western blot analysis confirmed that the expression of HA-ST14 in A549 and H1299 overexpressing stable transfectants was increased, indicating that the ST14 overexpressing cell line was successfully constructed. Figure 4 A), and then the CCK8 experimental results showed that the proliferation ability of ST14 cells overexpressed in A549 and H1299 decreased ( Figure 4 B), the results of clone formation experiments also showed that overexpression of ST14 in A549 and H1299 cells decreased cell proliferation ability ( Figure 4 C); Further, the role of ST14 was verified by constructing an ST14 knockdown cell line. Western blot confirmed that the expression of ST14 in A549 and H1299 knockdown stable cells was also reduced, indicating that the ST14 knockdown cell line was successfully constructed ( Figure 4 D), CCK8 experiments were performed, and the results showed that knocking down ST14 in A549 and H1299 cells enhanced cell proliferation ( Figure 4 E), the results of clone formation experiments also showed that knocking down ST14 in A549 and H1299 cells enhanced cell proliferation ( Figure 4 F); A subcutaneous tumor model in nude mice was then established using the A549 stable transgenic line overexpressing ST14. The results showed that the tumor weight and volume of cells overexpressing ST14 were significantly lower than those in the control group ( Figure 4 G- Figure 4 I), Ki67 immunohistochemical analysis of subcutaneous tumors showed overexpression of ST14 and decreased tumor cell proliferation energy ( Figure 4 J). In summary, these results indicate that ST14 promoter has the effect of inhibiting lung cancer cell proliferation and treating lung cancer.

[0038] In one embodiment of the present invention, we demonstrated that ST14 inhibits lung cancer cell proliferation by inhibiting O-GlcNAc glycosylation modification. Western blot analysis showed that compared with adjacent adjacent tissues, ST14 expression in lung cancer tissues was downregulated, and protein glycosylation levels were increased ( Figure 5 A), and then CCK8 test results showed that the proliferation ability of A549 and H1299 stably transfected cell lines overexpressing ST14 was restored by treating them with glycosyltransferase inhibitor TG ( Figure 5 B) Clone formation experiments further confirmed that the inhibitory effect of ST14 on the proliferation of A549 and H1299 cells could be reversed by glycosyltransferase inhibitors ( Figure 5 C); CCK8 assay results showed that the proliferation ability of ST14 knockdown A549 and H1299 stable cell lines was decreased when treated with the glycosylation inhibitor OGT-IN ( Figure 5 D) The clone formation experiment further confirmed that the promoting effect of ST14 knockdown on the proliferation of lung cancer cell lines could be reversed by OGT-IN ( Figure 5 D). Taken together, these results indicate that ST14 promoter exerts its effect by inhibiting O-GlcNAcylation modification.

[0039] In one embodiment of the present invention, we demonstrated that ST14 inhibits the binding of OGT to TKT and inhibits the O-GlcNAc glycosylation modification of TKT. First, we used A549 to perform ST14 binding mass spectrometry and O-GlcNAc glycosylation mass spectrometry analysis, and found that 319 proteins bound to ST14 were O-GlcNAc glycosylated ( Figure 6 A), for Figure 6 The 319 proteins in A were further enriched in Go and KEGG signaling pathways, and the results showed that glycolysis and pentose phosphate pathways were significantly changed ( Figure 6 B- Figure 6 C); Furthermore, Western blot analysis revealed that overexpression of ST14 in A549 and H1299 did not significantly change TKT expression, but O-GlcNAc-TKT expression was significantly downregulated ( Figure 6 D) When ST14 was knocked down, TKT expression did not change significantly, but O-GlcNAc-TKT expression increased ( Figure 6 E); Protein immunoprecipitation experiments confirmed that OGT and TKT can bind to each other ( Figure 6 F), glycosyltransferase inhibitor TG can promote TKT O-GlcNAc glycosylation modification ( Figure 6 G), knockdown of ST14 in A549 cells enhanced the binding of OGT to TKT ( Figure 6 H), Overexpression of ST14 in A549 cells weakened the binding of OGT to TKT ( Figure 6 I). Taken together, these results indicate that the ST14 promoter can negatively regulate the expression of O-GlcNAc-TKT.

[0040] In one embodiment of the present invention, we demonstrated that O-GlcNAc glycosylation of TKT can regulate the proliferation of lung cancer cells. SWGA experimental results showed that TKT 30,31 threonine and 345 serine affect its O-GlcNAc glycosylation ( Figure 7 A), TKT with three mutants of threonine at positions 30, 31 and serine at position 345 cannot undergo O-GlcNAc glycosylation modification ( Figure 7 B); Western blot further verified the expression of Flag-TKT WT and Flag-TKT mutants in cell lines, indicating that Flag-TKT mutants cannot undergo O-GlcNAc glycosylation modification ( Figure 7 C); CCK8 results showed that compared with wild-type TKT, the TKT glycosylation site mutant significantly weakened the promoting effect on cell proliferation ( Figure 7 D), clone formation experiments further confirmed that the promoting effect of TKT on lung cancer cell line proliferation depends on its O-GlcNAc glycosylation modification ( Figure 7 E); Figure 7 F shows that the promoting effect of TKT on glucose consumption in lung cancer cell lines is dependent on O-GlcNAc glycosylation modification. Figure 7 G shows that the promotion of lactate production by TKT in lung cancer cell lines is also dependent on O-GlcNAc glycosylation modification. Taken together, these results indicate that O-GlcNAc glycosylation modification of TKT can regulate the proliferation of lung cancer cells.

[0041] In one embodiment of the present invention, we demonstrated that the regulatory effect of ST14 on the proliferation and glycolysis pathway of lung cancer cell lines is dependent on TKT. Figure 8 A is the CCK8 result showing that overexpression of TKT under overexpression of ST14 can significantly reverse the inhibitory effect of ST14 on the proliferation of lung cancer cell lines; Figure 8 B. The results of clone formation experiments showed that overexpression of TKT under the condition of overexpression of ST14 could significantly reverse the inhibitory effect of ST14 on the proliferation of lung cancer cell lines; Figure 8 C shows that when ST14 is overexpressed and TKT is overexpressed, cellular glucose consumption is significantly increased; Figure 8 D shows the case of overexpression of ST14. Overexpression of TKT significantly increased the amount of cellular lactate production; Figure 8 E is the CCK8 result showing that knocking down TKT in the case of knocking down ST14 can significantly reverse the promoting effect of knocking down ST14 on the proliferation of lung cancer cell lines; Figure 8 F is the result of clone formation experiment showing that knocking down TKT in the case of knocking down ST14 can significantly reverse the promoting effect of knocking down ST14 on the proliferation of lung cancer cell lines; Figure 8 G shows the case of knocking down ST14. By knocking down TKT, cellular glucose consumption was significantly reduced; Figure 8 H shows the case of knockdown of ST14. Knocking down TKT significantly reduced cellular lactate production. Taken together, these results indicate that the regulatory effects of ST14 on lung cancer cell proliferation and glycolysis are dependent on TKT.

[0042] In one embodiment of the present invention, we demonstrated that the regulation of lung cancer cell proliferation and glycolysis by ST14 depends on the O-GlcNAc glycosylation modification of TKT. Figure 9 A is the CCK8 result showing that under the condition of double knockdown of ST14 and TKT, overexpression of TKT can reverse the promoting effect of ST14 knockdown on the proliferation of lung cancer cell lines, while overexpression of TKT glycosylation mutant has no obvious reversal effect. Figure 9 B is the result of clone formation experiment showing that under the condition of double knockdown of ST14 and TKT, overexpression of TKT can reverse the promoting effect of ST14 knockdown on the proliferation of lung cancer cell lines, while overexpression of TKT glycosylation mutant has no obvious reversal effect. Figure 9 C shows that in the case of ST14 and TKT double knockdown, overexpression of TKT promoted cellular glucose consumption and lactate production, while overexpression of a TKT glycosylation mutant had no significant promoting effect. Taken together, these results suggest that ST14 promoters can play a therapeutic role in lung cancer by regulating the expression level of O-GlcNAc-TKT.

[0043] 102: Processing data, inputting the O-GlcNAc-TKT expression data into a constructed evaluation model, wherein the evaluation model predicts the therapeutic effect of the ST14 promoter on lung cancer patients based on the O-GlcNAc-TKT expression data; In some embodiments, the method for constructing the evaluation model is known to those skilled in the art, and the step of associating the expression level of O-GlcNAc-TKT with a certain possibility or risk can be implemented and realized in different ways.

[0044] In the context of the present invention, the term "machine learning" refers to the use of computers to simulate or implement human learning activities. Technicians generally use different development tools to build machine learning algorithm models. The development tools include but are not limited to TensorFlow, Scikit Learn, PyTorch, OpenNN, RapidMiner, Azure Machine Learning, Apache Mahout, Shogun, KNIME, Vertex AI, H2Oai, Anaconda, Keras, Tableau, Fast.ai, Catalyst, Amazon ML, MLJAR, Spell. The algorithm models include but are not limited to linear regression models, logistic regression models, Lasso regression models, Ridge regression models, linear discriminant analysis models, nearest neighbor models, decision tree models, perceptron models, neural network models, support vector machine models, naive Bayes models, AdaBoost models, GBDT models, XGBoost models, LightGBM models, CatBoost models, or random forest models.

[0045] In one embodiment, after the evaluation model is constructed, the ROC curve analysis can be used to evaluate the effectiveness of the evaluation model.

[0046] An ROC curve is a plot of the true positive rate (sensitivity) of a test against the false positive rate (100% minus specificity) of the test. It is useful for depicting the performance of a particular feature when distinguishing between two populations. Typically, feature data is selected in ascending order across the entire population based on the value of a single feature. Then, for each value of that feature, the true positive and false positive rates of the data are calculated. The true positive rate is determined by counting the number of cases with a value above that feature and dividing it by the total number of cases. The false positive rate is determined by counting the number of controls with a value above that feature and dividing it by the total number of controls. While this definition refers to situations where a feature is elevated in cases compared to controls, it also applies to situations where a feature is low in cases compared to controls (in which case, samples with a value below that feature would be counted). ROC curves can be generated for individual features, as well as for other individual outputs. For example, a combination of two or more features can be mathematically combined (e.g., added, subtracted, multiplied, etc.) to provide a single summed value, which can then be plotted in an ROC curve. Additionally, any combination of multiple features where the combination is derived from a single output value can be plotted in an ROC curve.

[0047] 103: Output the result.

[0048] In the present invention, the evaluation model obtains results through the following criteria: when the expression level of the O-GlcNAc-TKT is lower than a threshold value, a classification result is obtained that the ST14 promoter is effective in treating lung cancer patients; when the expression level of the O-GlcNAc-TKT is higher than a threshold value, a classification result is obtained that the ST14 promoter is ineffective in treating lung cancer patients.

[0049] In some embodiments, the preset threshold is a representative value of normal samples from a lung cancer population, including but not limited to a maximum value, a third quartile, and an average value. In some preferred embodiments of the present invention, the population sample includes more than 20 samples, for example, 30, 50, 80, 100, 150, 200, 300, 500, or more.

[0050] Figure 2 A schematic diagram of a system for evaluating the therapeutic effect of ST14 promoter on lung cancer provided by an embodiment of the present invention; The system is programmed or otherwise configured to include a data acquisition unit 201, a data classification unit 202, and a result output unit 203: Data acquisition unit 201: used to obtain the expression data of O-GlcNAc-TKT in lung cancer patient samples after ST14 promoter treatment; Data classification unit 202: used to classify and predict the data obtained by the data acquisition unit using the evaluation model obtained by the construction method described in the first aspect of the present invention, and obtain a classification result of whether the ST14 promoter is effective in treating lung cancer patients; Result output unit 203: used to output classification results.

[0051] The system may be the user's electronic device or a computer system located remotely from the electronic device.

[0052] Figure 3 This is a schematic structural diagram of the computer device provided by the present invention.

[0053] The computer device 300 includes a processor 301 and a memory 302 coupled to the processor 301 . The memory 302 stores program instructions. When the program instructions are executed by the processor 301 , the processor 301 executes the above-mentioned method for evaluating the therapeutic effect of ST14 promoter on lung cancer.

[0054] The processor 301 may also be referred to as a CPU (Central Processing Unit). The processor 301 may be an integrated circuit chip having signal processing capabilities. The processor 301 may also be a general-purpose processor, a digital signal processor (DSP), an application-specific integrated circuit (ASIC), a field-programmable gate array (FPGA), or other programmable logic device, a discrete gate or transistor logic device, or a discrete hardware component. The general-purpose processor may be a microprocessor or any conventional processor.

[0055] The computer device 300 may be a mobile electronic device.

[0056] It should be understood that the systems, devices, and methods described herein can be implemented in other ways. For example, the device embodiments described above are merely illustrative. For example, the module division is merely a logical functional division, and actual implementations may employ other division methods, such as combining or integrating multiple modules or components into another system, or omitting or disabling certain features. Furthermore, the coupling or direct coupling or communication connection shown or discussed may be an indirect coupling or communication connection between devices or modules via some interface, which may be electrical, mechanical, or other forms.

[0057] The modules described as separate components may or may not be physically separate, and the components shown as modules may or may not be physical units, that is, they may be located in one place or distributed across multiple network elements. Some or all of these units may be selected to achieve the purpose of this embodiment according to actual needs.

[0058] In addition, the functional modules in various embodiments of the present invention may be integrated into a single processing module, or each module may exist physically separately, or two or more modules may be integrated into a single module. The aforementioned integrated modules may be implemented in the form of hardware or software functional modules.

[0059] The above is only an implementation method of the present application and does not limit the patent scope of the present application. Any equivalent structure or equivalent process transformation made using the contents of the description and drawings of this application, or directly or indirectly applied in other related technical fields, are also included in the patent protection scope of the present application.

Claims

1. A method for evaluating the therapeutic effect of ST14 promoter on lung cancer, characterized in that: The method is performed by a computer and comprises the following steps: Obtain data: Obtain O-GlcNAc-TKT expression data in lung cancer patient samples after ST14 promoter treatment; Processing data: inputting the O-GlcNAc-TKT expression data into a constructed evaluation model, wherein the evaluation model predicts the therapeutic effect of the ST14 promoter on lung cancer patients based on the O-GlcNAc-TKT expression data; Output the results.

2. The method according to claim 1, characterized in that The steps for constructing the evaluation model are as follows: Obtaining O-GlcNAc-TKT expression data from untreated lung cancer patients and lung cancer patients treated with an ST14 promoter; inputting the O-GlcNAc-TKT expression data into a machine learning algorithm to construct an evaluation model.

3. The method according to claim 2, characterized in that The machine learning algorithm includes an algorithm model developed using various development tools; Preferably, the development tools include but are not limited to TensorFlow, Scikit Learn, PyTorch, OpenNN, RapidMiner, Azure Machine Learning, Apache Mahout, Shogun, KNIME, Vertex AI, H2Oai, Anaconda, Keras, Tableau, Fast.ai, Catalyst, Amazon ML, MLJAR, Spell; Preferably, the algorithm model includes but is not limited to a linear regression model, a logistic regression model, a Lasso regression model, a Ridge regression model, a linear discriminant analysis model, a nearest neighbor model, a decision tree model, a perceptron model, a neural network model, a support vector machine model, a naive Bayes model, an AdaBoost model, a GBDT model, an XGBoost model, a LightGBM model, a CatBoost model, and a random forest model.

4. The method according to claim 1, wherein The evaluation model obtains results through the following criteria: When the expression level of the O-GlcNAc-TKT is lower than the threshold, a classification result is obtained that the ST14 promoter is effective in treating lung cancer patients; when the expression level of the O-GlcNAc-TKT is higher than the threshold, a classification result is obtained that the ST14 promoter is ineffective in treating lung cancer patients.

5. A system for evaluating the therapeutic effect of ST14 promoter on lung cancer, characterized in that: The system comprises: Data acquisition unit: used to obtain the expression data of O-GlcNAc-TKT in lung cancer patient samples after ST14 promoter treatment; A data classification unit: configured to classify and predict the data obtained by the data acquisition unit using the evaluation model obtained by the construction method according to claim 2, and obtain a classification result of whether the ST14 promoter is effective in treating lung cancer patients; Result output unit: used to output classification results.

6. A computer device, characterized in that: The computer device includes a memory and a processor, the memory is used to store program instructions; the processor is used to call the program instructions, and when the program instructions are executed, the method for evaluating the therapeutic effect of ST14 promoter on lung cancer according to any one of claims 1-4 is implemented.

7. A computer-readable storage medium, characterized in that The computer-readable storage medium stores a computer program, and when the computer program is executed by a processor, the method for evaluating the therapeutic effect of an ST14 promoter on lung cancer according to any one of claims 1 to 4 is implemented.

8. A method for regulating the expression of O-GlcNAc-TKT in lung cancer cells, characterized in that: In vitro, ST14 negatively regulates the expression level of O-GlcNAc-TKT in lung cancer cells; by regulating the expression level of ST14, the expression of O-GlcNAc-TKT in lung cancer cells can be regulated.

9. The method according to claim 8, characterized in that Upregulating ST14 expression decreased O-GlcNAc-TKT expression; inhibiting ST14 expression increased O-GlcNAc-TKT expression; Preferably, the upregulation of ST14 expression is achieved by gene overexpression technology; Preferably, the gene overexpression vector includes a eukaryotic expression vector and a viral vector.

10. Application of ST14 promoter in the preparation of drugs for treating lung cancer; Preferably, the ST14 promoter acts by inhibiting the expression level of O-GlcNAc-TKT.

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