Use of ttll1 inhibitors in the preparation of a medicament for alleviating and / or treating liver cancer

Abstract

The application discloses application of a TTLL1 inhibitor in preparation of a medicine for relieving and / or treating liver cancer, and belongs to the field of biomedical technology. The application discloses application of a TTLL1 expression inhibitor in preparation of a medicine for relieving and / or treating liver cancer, and the expression of TTLL1 is down-regulated through the TTLL1 expression inhibitor, so that the proliferation activity and migration ability of liver cancer cells can be effectively inhibited. The application further provides a complete scheme for preparing various dosage forms of medicines by using the TTLL1 expression inhibitor, and the technical scheme has good clinical transformation prospect and application potential as a brand-new anti-liver cancer medicine.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to the application of TTLL1 inhibitors in the preparation of drugs for alleviating and / or treating liver cancer. Background Technology

[0002] Hepatocellular carcinoma (HCC) is the most prevalent histological type of primary liver cancer, accounting for 70%-85% of all cases. Currently, the preferred treatment for liver cancer is early surgical resection. However, due to the insidious nature of early-stage symptoms, most patients are diagnosed at an intermediate or advanced stage, thus losing the opportunity for radical surgery. The natural course of the disease in these patients is typically only 3-6 months, and they can only receive palliative care or systemic chemotherapy, but the overall efficacy is limited and the prognosis is poor. Therefore, there is an urgent need to develop new treatment strategies and targets.

[0003] It should be noted that although both hepatocellular carcinoma and gastric cancer are common malignant tumors of the digestive system, they differ significantly in their specific causes, mechanisms of development, tumor microenvironment, and clinical treatment. The occurrence of hepatocellular carcinoma is usually closely related to chronic hepatitis B virus infection, hepatitis C virus infection, long-term alcohol intake, non-alcoholic fatty liver disease, aflatoxin exposure, and a background of cirrhosis. Its pathogenesis often follows a continuous pathological evolution of "chronic hepatitis—liver fibrosis—cirrhosis—liver cancer." In contrast, the occurrence of gastric cancer is more related to Helicobacter pylori infection, chronic atrophic gastritis, intestinal metaplasia, gastric mucosal dysplasia, dietary factors, and genetic susceptibility. Its typical evolutionary process often manifests as "chronic gastritis—atrophic gastritis—intestinal metaplasia—dysplasia—gastric cancer." Therefore, liver cancer and gastric cancer have different tissue origins, underlying disease backgrounds, and precancerous lesion evolution pathways.

[0004] From a pathogenesis perspective, hepatocellular carcinoma typically occurs in the context of long-term chronic liver injury and liver regeneration. Hepatocytes repeatedly undergo necrosis, regeneration, and abnormal proliferation under continuous inflammatory stimulation, oxidative stress, liver fibrosis, and the cirrhotic microenvironment, leading to genomic instability, cell cycle dysregulation, enhanced angiogenesis, and increased invasiveness and metastasis. The liver itself possesses unique metabolic functions, detoxification functions, blood supply characteristics, and an immune tolerance microenvironment, giving hepatocellular carcinoma its own characteristics in terms of molecular regulatory networks, tumor angiogenesis, immune escape, and treatment tolerance. Gastric cancer, on the other hand, primarily originates from gastric mucosal epithelial cells, and its development is closely related to gastric mucosal barrier disruption, chronic inflammation, abnormal epithelial cell differentiation, altered cell adhesion, destruction of gastric gland structure, and changes in the local gastric mucosal microenvironment. Therefore, hepatocellular carcinoma is not simply a different manifestation of gastric cancer in different organs, but rather a type of malignant tumor with independent etiological basis, histological origin, and molecular pathological characteristics.

[0005] In terms of treatment options, hepatocellular carcinoma (HCC) and gastric cancer also differ significantly. Treatment choices for HCC depend not only on tumor size, number, vascular invasion, and distant metastasis, but also heavily on factors such as the patient's liver function reserve, degree of cirrhosis, and portal hypertension. Clinical treatment methods include liver resection, liver transplantation, radiofrequency ablation, microwave ablation, transarterial chemoembolization, radiotherapy, targeted therapy, immunotherapy, and their combination therapies. In contrast, gastric cancer treatment typically centers on gastrectomy and lymph node dissection, combined with perioperative chemotherapy, radiotherapy, targeted therapy, and immunotherapy. Treatment decisions are more focused on factors such as the depth of gastric wall invasion, lymph node metastasis, distant metastasis, HER2 status, microsatellite instability, and PD-L1 expression. Because HCC patients often have underlying liver disease and impaired liver function, drug metabolism, treatment tolerance, local treatment methods, and prognostic assessments differ significantly from those for gastric cancer. Therefore, even if certain molecular targets or intervention methods may have superficial similarities in different tumors, their specific application basis, indications, action environment, and therapeutic significance in hepatocellular carcinoma still have significant differences.

[0006] The TTLL1 (tubulin tyrosine ligase-like 1) gene is a protein-encoding gene, also known as the TTL family tubulin polyglutamylase complex subunit L1. It belongs to the TTL-like protein family and encodes a tubulin polyglutamylase complex-associated protein. TTLL1 primarily participates in the post-translational modification of tubulin, catalyzing or promoting polyglutamylation of tubulin, thereby regulating various biological processes such as microtubule structural stability, microtubule dynamics, cytoskeleton remodeling, and intracellular transport. The microtubule system is a crucial structural basis for maintaining cell morphology, cell polarity, chromosome segregation, mitosis, and cell migration. Tubulin polyglutamylation, as an important form of post-translational modification, affects the interaction between microtubules and motor proteins, microtubule-associated proteins, and other regulatory factors, playing a vital regulatory role in cell division, cell motility, and signal transduction. Abnormal TTLL1 function can lead to an imbalance in tubulin modification, thereby affecting cell cycle progression, cytoskeleton homeostasis, chromosome segregation, and cell migration and invasion capabilities.

[0007] Existing research indicates that abnormal microtubule structure and dynamics are key molecular characteristics of tumorigenesis and development. During rapid proliferation, invasion, metastasis, and treatment resistance, tumor cells often exhibit cytoskeletal remodeling, abnormal spindle assembly, increased chromosomal instability, and enhanced cell migration. The microtubule system is also a crucial target for many classic antitumor drugs; for example, taxanes and vinca alkaloids can inhibit tumor cell proliferation by interfering with microtubule polymerization or depolymerization. As a key molecule regulating tubulin polyglutamylation, although studies have revealed the basic biological functions of TTLL1 from the perspectives of tubulin polyglutamylation, cytoskeleton regulation, and cell motility, the specific role and molecular mechanisms of TTLL1 in the development and progression of tumors, especially solid tumors such as hepatocellular carcinoma, remain unclear. Currently, there is a lack of systematic research exploring the relationship between TTLL1-mediated microtubule modification abnormalities, cytoskeleton homeostasis changes, and hepatocellular carcinoma cell proliferation, invasion, metastasis, and treatment sensitivity. Furthermore, there is a lack of intervention strategies and translational applications targeting TTLL1. Therefore, further research on the function of TTLL1 and its role in liver cancer, especially the development of TTLL1 inhibitors for the preparation of drugs to alleviate and / or treat liver cancer, has important scientific significance and potential clinical value.

[0008] Therefore, developing a drug that targets TTLL1 to alleviate and / or treat liver cancer has become an urgent need in this field. Summary of the Invention

[0009] The purpose of this invention is to explore the specific role of TTLL1 in hepatocellular carcinoma and to provide a drug that targets TTLL1 to alleviate and / or treat liver cancer.

[0010] In a first aspect, the present invention provides the use of a TTLL1 expression inhibitor in the preparation of drugs for alleviating and / or treating liver cancer, wherein the TTLL1 expression inhibitor is shRNA; the nucleotide sequence of the shRNA is shown in SEQ ID NO: 1 and SEQ ID NO: 2.

[0011] In a second aspect, the present invention provides the use of a drug containing a TTLL1 expression inhibitor in the preparation of a drug for alleviating and / or treating liver cancer, wherein the TTLL1 expression inhibitor is a lentivirus comprising shRNA, the nucleotide sequence of which is shown in SEQ ID NO: 1 and SEQ ID NO: 2.

[0012] Preferably, the drug further includes pharmaceutically acceptable excipients; the dosage form of the drug is an injection.

[0013] This invention retrieved and compared the expression differences of TTLL1 in hepatocellular carcinoma (HCCDB) tissues, adjacent normal tissues, cirrhotic tissues, and normal liver tissues using the Hepatocellular Carcinoma Expression Atlas Database (HCCDB). The results showed that the mRNA expression level of TTLL1 in HCC tissues was significantly higher than that in the corresponding adjacent normal tissues, cirrhotic tissues, and normal liver tissues. Using the TCGA-LIHC dataset (Cancer Genome Atlas Project - Hepatocellular Carcinoma) from the Solid Tumor Biomarker Exploration Platform (BEST), the relationship between TTLL1 expression levels and the prognosis of HCC patients was analyzed. The results showed that the survival rate of patients in the high TTLL1 expression group was significantly lower than that of the low TTLL1 expression group. Based on these bioinformatics analyses, TTLL1 was identified as a potential diagnostic biomarker and therapeutic target for HCC.

[0014] This invention uses human hepatocellular carcinoma (HLC) cells as experimental subjects. Through knockdown experiments, it verifies the effect of shRNA lentiviruses targeting TTLL1 on the expression level of TTLL1 protein, verifies the effect of TTLL1 inhibitors on the migration and proliferation of hepatocellular carcinoma cells, and confirms the effect of TTLL1 inhibitors on the activity of human HLC cells.

[0015] Compared with the prior art, the present invention has the following advantages: This invention, through large-scale bioinformatics analysis of clinical samples, demonstrates that the TTLL1 gene is specifically highly expressed in hepatocellular carcinoma tissues, and that this high expression is significantly associated with poor patient prognosis. This reveals that the TTLL1 gene is a key factor in hepatocellular carcinoma, suggesting that TTLL1 could serve as a potential diagnostic biomarker and therapeutic target for liver cancer.

[0016] This invention experimentally validates that TTLL1 expression inhibitors can serve as drugs for alleviating and / or treating liver cancer. Specifically, by specifically downregulating TTLL1 gene expression through lentiviruses carrying shRNA, significant anti-cancer effects can be generated in multiple aspects in an in vitro model.

[0017] This invention not only provides the theoretical basis for TTLL1 expression inhibitors as therapeutic drugs for liver cancer, but also provides a complete scheme for preparing drugs in various dosage forms. These technical solutions, as a novel class of anti-liver cancer drugs, have good prospects for clinical translation and application potential. Attached Figure Description

[0018] Figure 1 This is a graph showing the results of analyzing the differences in TTLL1 expression in liver cancer tissue, adjacent tissue, cirrhotic tissue, and normal liver tissue using the dataset in Example 1. Figure 2 This is a graph showing the relationship between TTLL1 expression level and overall survival, disease-specific survival, progression-free survival, and disease-free survival in Example 1, based on the TCGA-LIHC dataset. Figure 3 This is a graph showing the results of Western blot verification of the effect of TTLL1 inhibitor on TTLL1 protein expression level in HLF liver cancer cells in Example 2; Figure 4 This is a scratch healing experiment diagram used in Example 3 to detect the effect of TTLL1 inhibitors on the migration ability of liver cancer cells; Figure 5 This is a CCK-8 assay diagram used in Example 4 to detect the effect of TTLL1 inhibitors on the proliferation of liver cancer cells. Figure 6 This is an EdU probe labeling experiment used in Example 4 to detect the effect of TTLL1 inhibitors on the proliferation of liver cancer cells. Detailed Implementation

[0019] To better understand the present invention, the following embodiments are further illustrations of the invention, but the scope of the invention is not limited to these embodiments. Unless otherwise specified, the experimental methods used in the embodiments of the present invention are conventional methods. The reagents and materials used in the embodiments are all commercially available, and the quantitative experiments involved in the embodiments are all performed in at least three replicates.

[0020] Example 1: Bioinformatics Analysis of TTLL1 Expression and Prognostic Value in Hepatocellular Carcinoma The expression levels of TTLL1 in datasets GSE22058, GSE25097, GSE36376, GSE14520, GSE46444, GSE54236, GSE63898, TCGA-LIHC, ICGC-LIRI-JP, GSE87630, GSE76427, OEP000321, GSE121248, GSE89377, and GSE148355 were retrieved and analyzed using the HCCDB database (http: / / lifeome.net:809 / # / home). The expression differences of TTLL1 in hepatocellular carcinoma tissue, adjacent non-cancerous tissue, cirrhotic tissue, and normal liver tissue were compared.

[0021] The relationship between TTLL1 expression level and prognosis in hepatocellular carcinoma (HCC) patients in the TCGA-LIHC dataset was analyzed using the BEST database (https: / / rookieutopia.hiplot.com.cn / app_direct / BEST / #PageHomeAnalysisModuleSelection). Patients were divided into a high TTLL1 expression group and a low TTLL1 expression group using the median TTLL1 expression level as the grouping cutoff. Kaplan-Meier curves were plotted for overall survival, disease-specific survival, progression-free survival, and disease-free survival, and the Log-rank test was used to compare survival differences between the two groups. Statistical significance was indicated as follows: P<0.05; P<0.01; P < 0.001; when P < 0.05, the difference is considered statistically significant.

[0022] The results of the analysis of TTLL1 expression differences in hepatocellular carcinoma tissues, adjacent non-cancerous tissues, cirrhotic tissues, and normal liver tissues based on the datasets GSE22058, GSE25097, GSE36376, GSE14520, GSE46444, GSE54236, GSE63898, TCGA-LIHC, ICGC-LIRI-JP, GSE87630, GSE76427, OEP000321, GSE121248, GSE89377, and GSE148355 are shown in the figure below. Figure 1 As shown in the figure, it can be seen that in all 15 datasets included, the mRNA expression level of TTLL1 in liver cancer tissues was significantly higher than that in the corresponding adjacent normal tissues, cirrhotic tissues and normal liver tissues (P<0.05).

[0023] The relationship between TTLL1 expression levels and overall survival, disease-specific survival, progression-free survival, and disease-free survival based on the TCGA-LIHC dataset is shown in the figure below. Figure 2 As shown in the figure, in the TCGA-LIHC dataset, the overall survival, disease-specific survival, progression-free survival, and disease-free survival of patients in the TTLL1 high expression group were significantly shorter than those in the low expression group (Log-rank P<0.05).

[0024] In summary, TTLL1 is highly expressed in hepatocellular carcinoma tissues, and high TTLL1 expression is associated with poor prognosis in hepatocellular carcinoma patients, suggesting that TTLL1 may serve as a potential diagnostic biomarker and therapeutic target for hepatocellular carcinoma.

[0025] Example 2: Construction of shRNA lentiviral vector targeting TTLL1 and establishment of stable knockdown cell lines 1. shRNA sequence design and synthesis A specific shRNA interference fragment was designed targeting the human TTLL1 (Gene ID: 25809) mRNA sequence. Its template sequence is as follows: Justice Chain (SEQ ID NO: 1): GTTTGTGTCTCAATCTAATAAttcaagagaTTATTAGATTGAGACACAAAC; Antisense chain (SEQ ID NO: 2): CAAACACAGAGTTAGATTATTaagttctctAATAATCTAACTCTGTGTTTG.

[0026] 2. Construction of lentiviral vectors After annealing the synthesized double-stranded oligonucleotides to form double strands, they were cloned into the pLV3-U6-MCS-shRNA-CopGFP-Puro lentiviral vector (Wuhan Miaoling Biotechnology Co., Ltd., catalog number P29436) which had been double-digested with AgeI and EcoRI, to construct the recombinant plasmid pLV3-U6-TTLL1(human)-shRNA1-CopGFP-Puro (Wuhan Miaoling Biotechnology Co., Ltd., catalog number P105476). The correctness of the inserted sequence was verified by colony PCR and sequencing.

[0027] 3. Lentiviral packaging The virus was packaged using a three-plasmid system. Healthy 293T cells were seeded in 10 cm culture dishes. When the cell density reached 70%-80%, the following plasmids were co-transfected using liposome transfection: 8 μg of pLV3-U6-TTLL1(human)-shRNA1-CopGFP-Puro (or the empty control plasmid pLV3-U6-MCS-shRNA-CopGFP-Puro), 6 μg of the packaging plasmid psPAX2, and 2 μg of the envelope plasmid pMD2.G. Six hours after transfection, the medium was replaced with complete medium (DMEM liquid medium + 10% fetal bovine serum FBS). After 48 h and 72 h of culture, the virus-containing supernatant was collected, filtered through a 0.45 μm filter, and the virus was concentrated by ultracentrifugation to obtain lentiviral particles (i.e., the TTLL1 inhibitor), which were then stored at -80℃.

[0028] 4. Cell infection and screening Human hepatocellular carcinoma HLF cells were seeded in 6-well plates (5 × 10⁻⁶ cells / well). 5Cells were cultured in / wells until the density reached approximately 30%. Viral supernatant (MOI=10) and polybrene transfection reagent at a final concentration of 5 μg / mL were added. After 24 h of infection, the medium was replaced with fresh complete medium and cultured for another 48 h. Puromycin was then added at a final concentration of 2 μg / mL for selection, continuing for approximately one week until all untransfected control cells died. Monoclonal cell lines were obtained using limiting dilution, and green fluorescence expression was observed under a fluorescence microscope to assess infection efficiency. Finally, a stable TTLL1 knockdown shTTLL1 cell line and an empty vector control shNC cell line were obtained.

[0029] 5. Low-efficiency verification Total protein was extracted from shTTLL1 and shNC cells and Western blot was performed to verify the knockdown effect of TTLL1 protein.

[0030] Prepare the protein extraction buffer according to the following ratio: RIPA lysis buffer: 1 mL; 50× protease inhibitor: 20 μL; 100× phosphatase inhibitor A: 10 μL; 100× phosphatase inhibitor B: 10 μL; 100× PMSF protease inhibitor: 10 μL. Mix well and use on ice.

[0031] Discard the culture medium and wash with PBS; add protein extraction buffer to lyse the cells for 5 min and collect with a scraper; then sonicate to disrupt the cells, and incubate the lysate on ice for 10 min. Centrifuge at 12000 rpm and 4℃ for 10 min, and collect the supernatant as total cell protein.

[0032] Mix total cellular protein with 5× loading buffer at a ratio of 4:1, and heat at 100℃ for 5 min to denature the protein; the protein sample can be directly used for electrophoresis after cooling to room temperature.

[0033] Place the pre-prepared SDS-PAGE gel into the electrophoresis tank and add electrophoresis buffer; add protein samples and pre-stained molecular weight markers to the upper sample wells. Electrophoresis is performed at 80 V, and after the samples enter the separating gel, the voltage is adjusted to 120 V. Electrophoresis is stopped when the bromophenol blue reaches near the bottom of the gel.

[0034] Cut the polyvinylidene fluoride (PVDF) membrane and activate it with methanol before placing it in the transfer buffer. Assemble the gel and PVDF membrane in the order of sponge-filter paper-gel-membrane-filter paper-sponge and place it in a wet transfer apparatus. Transfer the membrane at 4°C and 200 mA for 60 min.

[0035] After transfer, the membrane was washed with TBST buffer and blocked with TBST buffer containing 10% skim milk powder for 2 hours. Then, diluted primary antibody (TTLL1 Polyclonal Antibody, Invitrogen, catalog number PA5-27285) was added and incubated overnight at 4°C. After washing, HRP-labeled secondary antibody was added and incubated at room temperature for 1 hour, followed by another wash. The membrane was then placed in an ECL chemiluminescent substrate for development and exposed in an imaging system to acquire band images.

[0036] Western blotting results are as follows: Figure 3 As shown, compared with the shNC group, the expression level of TTLL1 protein in shTTLL1 cells was significantly inhibited (P<0.01), and the expression level of internal control GAPDH was similar in both groups, indicating that this embodiment successfully constructed a shRNA lentivirus targeting TTLL1 and established a stable TTLL1 knockdown liver cancer cell model.

[0037] Example 3: Effect of TTLL1 inhibitors on the migration ability of liver cancer cells The shTTLL1 and shNC cells from Example 2 were seeded into 6-well plates and cultured until cell confluence reached over 90%. Using a 200 μL sterile pipette tip, three straight scratches were made perpendicular to the bottom of each well in the monolayer. The cells were washed 2-3 times with PBS to remove cell debris. The medium was then replaced with serum-free medium to inhibit cell proliferation. The scratches at 0 h were photographed and marked under a microscope (100x). After culturing the plates for another 48 h, the same locations were photographed again. The width of the scratches was measured using ImageJ software, and the scratch healing rate was calculated using the following formula: Healing rate (%) = [(0 h scratch width - 48 h scratch width) / 0 h scratch width] × 100% Experimental results and migration rate comparison charts showing the effect of TTLL1 inhibitors on the migration ability of liver cancer cells, as shown in the figure. Figure 4 As shown, after 48 h of culture, the scratch healing rate of the shTTLL1 group was significantly lower than that of the shNC group (P<0.05), indicating that the TTLL1 inhibitor can significantly inhibit the in vitro migration ability of liver cancer cells.

[0038] Example 4: Effect of TTLL1 inhibitors on the proliferation of liver cancer cells 1. CCK-8 Experiment shNC and shTTLL1 cells from Example 2 were cultured until they reached the logarithmic growth phase and seeded into 96-well plates at a density of 3000 cells per well, with 5 replicates per group. At 24 h, 48 h, and 72 h post-seeding, 10 μL of CCK-8 reagent (Biosharp) was added to each well, and the plates were incubated at 37°C for 2 h. The absorbance (OD value) of each well was measured using a microplate reader at 450 nm. Cell proliferation curves were plotted with time on the x-axis and OD value on the y-axis.

[0039] Cell proliferation curve as shown Figure 5 As shown, at the 72-hour time point, the optical density OD value of the shTTLL1 group was significantly lower than that of the shNC group (P<0.001), indicating that the TTLL1 inhibitor significantly inhibited the proliferation of liver cancer cells.

[0040] 2. EdU cell proliferation detection The shNC and shTTLL1 cells from Example 2 were seeded into 24-well plates and cultured to a density of approximately 50%. (According to BeyoClick...) TM Following the instructions of the EdU-555 cell proliferation assay kit, add culture medium containing 10 μM EdU to each well and incubate for 2 h. Discard the culture medium, wash with PBS, fix with 4% paraformaldehyde for 15 min, and permeabilize with 0.3% Triton X-100 for 15 min. Add Click reaction solution (protected from light) and incubate for 30 min, then stain the cell nuclei with Hoechst 33342. After washing with PBS, observe under a fluorescence microscope and randomly photograph multiple fields of view.

[0041] Nuclear staining results as follows Figure 6 As shown, the EdU-positive cell rate in the shTTLL1 group was lower than that in the shNC group, indicating that the TTLL1 inhibitor can suppress the DNA replication activity of liver cancer cells. Combined with the CCK-8 assay results, this demonstrates that the TTLL1 inhibitor can significantly inhibit the in vitro proliferation ability of liver cancer cells.

[0042] Example 5: Injectable form of TTLL1 inhibitor Take 1 mL of the TTLL1 inhibitor (titer ≥ 1 × 10⁸ IFU / mL) prepared in Example 2, add PBS buffer (pH = 7.4) containing 5% sucrose and 1% human serum albumin to 10 mL, filter aseptically through a 0.22 μm microporous membrane, dispense into vials, and freeze to obtain the TTLL1 inhibitor injection.

[0043] Example 6: Lyophilized Powder Injection of TTLL1 Inhibitor Take the TTLL1 inhibitor injection from Example 5, dispense 1 mL per vial, place in a freeze dryer, pre-freeze, dry once, dry twice, and then cap and seal to obtain the lyophilized powder injection of the TTLL1 inhibitor. Reconstitute with physiological saline before use.

[0044] Example 7: Lipid nanoparticle (LNP) formulation of TTLL1 inhibitor The shRNA targeting TTLL1 with nucleotide sequences as shown in SEQ ID NO: 1 and SEQ ID NO: 2 in Example 2 was used as a TTLL1 inhibitor. It was mixed with ionizable lipids, phospholipids, cholesterol and PEG-lipids, and LNP formulations loaded with TTLL1 inhibitors were prepared by microfluidic technology. It can be used for intravenous injection to improve in vivo delivery efficiency.

[0045] SEQ ID NO:1 shRNA positive chain GTTTGTGTCTCAATCTAATAAttcaagagaTTATTAGATTGAGACACAAAC SEQ ID NO:2 shRNA antisense strand CAAACACAGAGTTAGATTATTaagttctctAATAATCTAACTCTGTGTTTG Nucleotide sequence of SEQ ID NO:3 TTLL1 The amino acid sequence of SEQ ID NO:4 TTLL1 MAGKVKWVTDIEKSVLINNFEKRGWVQVTENEDWNFYWMSVQTIRNVFSVEAGYRLSDDQIVNHFPNHYELTRKDLMVKNIKRYRKELEKEGSPLAEKDENGKYL YLDFVPVTYMLPADYNLFVEEFRKSPSSTWIMKPCGKAQGKGIFLINKLSQIKKWSRDSKTSSFVSQSNKEAYVISLYINNPLLIGGRKFDLRLYVLVSTYRPLRC YMYKLGFCRFCTVKYTPSTSELDNMFVHLTNVAIQKHGEDYNHIHGGKWTVSNLRLYLESTRGKEVTSKLFDEIHWIIVQSLKAVAPVMNNDKHCFECYGYDIIID DKLKPWLIEVNASPSLTSSTANDRILKYNLINDTLNIAVPNGEIPDCKWNNKSPPKEVLGNYEILYDEELAQGDGADRELRSRQGQSLGPRAGRSRDSGRAVLTTWK The above description is merely a preferred embodiment of the present invention, and should not be construed as limiting the scope of the invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principles of the present invention, and these improvements and modifications are also considered to be within the scope of protection of the present invention.

Claims

1. The application of TTLL1 expression inhibitors in the preparation of drugs for alleviating and / or treating liver cancer, characterized in that, The TTLL1 expression inhibitor is shRNA; the nucleotide sequence of the shRNA is shown in SEQ ID NO: 1 and SEQ ID NO:

2.

2. The use of a drug containing a TTLL1 expression inhibitor in the preparation of a drug for alleviating and / or treating liver cancer, characterized in that, The TTLL1 expression inhibitor is a lentivirus comprising shRNA, the nucleotide sequence of which is shown in SEQ ID NO: 1 and SEQ ID NO:

2.

3. The application according to claim 2, characterized in that, The drug also includes pharmaceutically acceptable excipients; the drug is in the form of an injection.

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