Application of LncRNA-ZFHX4-AS1 in the treatment and evaluation of adrenal tumors

Abstract

The present invention relates to the application of LncRNA-ZFHX4-AS1 in the treatment and evaluation of adrenal tumors. Through a large number of studies, the present invention found that LncRNA-ZFHX4-AS1 is a gene that is extremely relevant to the occurrence and development of adrenal tumors; by inhibiting the expression of LncRNA-ZFHX4-AS1, the proliferation, cloning, invasion and other activities and functions of adrenal tumors can be significantly inhibited; a higher expression level of LncRNA-ZFHX4-AS1 is significantly positively correlated with adrenal tumors, which can be used to predict the malignancy and treatment prognosis of adrenal tumor diseases, and provide early assessment for patients receiving preoperative treatment. The present invention provides a new drug treatment target for humans to conquer adrenal tumors, thereby providing a new direction for subsequent drug research and development, clinical treatment, etc., and has extremely high social value and market application prospects.

Description

Technical Field

[0001] The present invention belongs to the field of biomedical technology and relates to the application of LncRNA-ZFHX4-AS1 in the treatment and evaluation of adrenal tumors. Background Art

[0002] Adrenocortical Carcinoma (ACC) is a rare malignant endocrine tumor. According to statistics, the annual incidence of ACC in adults is 0.5-2 cases per million people. The early symptoms of adrenal tumors are hidden, and most patients have already invaded locally or metastasized to distant sites when they seek medical treatment, resulting in a poor prognosis. The five-year overall survival rate is 37-59%. Currently, radical surgery is still the only effective cure for local ACC. However, even if the tumor is completely removed, up to 25% of ACC patients will experience local recurrence or distant metastasis. For these ACC patients with a higher risk of recurrence or metastasis, more active and reliable diagnosis and treatment plans will be adopted clinically to improve the patient's prognosis, including chemotherapy, radiotherapy and targeted therapy. Accurate risk stratification and prognosis prediction for ACC patients remain key issues that urgently need to be addressed in current clinical diagnosis and treatment.

[0003] In recent years, indicators such as tumor proliferation index (Ki67), tumor stage, and whether the surgical margin of the tumor is positive have been widely used to predict the prognosis of ACC patients. However, due to the influence of tumor heterogeneity, these clinical indicators are gradually unable to meet the needs of personalized diagnosis and treatment and prognosis assessment of ACC patients. With the rapid development of technologies such as high-throughput sequencing, long noncoding RNA (lncRNA) with complex and precise regulatory functions has gradually received attention in the biological and medical fields. At present, a large number of studies have revealed that lncRNA plays an indispensable role in many biological processes such as the occurrence, development, deterioration and metastasis of human malignant tumors. LncRNA ZFHX4 Antisense RNA 1 (ZFHX4-AS1) was first discovered to be associated with 8q21.11 deletion syndrome, but its related research in ACC has not been reported. Summary of the Invention

[0004] The purpose of the present invention is to address the technical problems in the prior art of adrenal tumors, such as unclear etiology, difficulty in early prevention, ineffective treatment, and poor prognosis, thereby providing a new therapeutic target and strategy. By using LncRNA-ZFHX4-AS1 as a key target, it can effectively perform auxiliary diagnosis, treatment, and prognosis assessment of adrenal tumors.

[0005] In order to achieve the above object, the technical solution adopted by the present invention is:

[0006] A first aspect of the present invention provides the use of LncRNA-ZFHX4-AS1 in the preparation of a product for auxiliary diagnosis, treatment and / or prognosis evaluation of adrenal tumors.

[0007] Preferably, the adrenal tumor is selected from one or more of adrenal cortical tumor, adrenal medullary tumor, adrenal stromal tumor and adrenal metastasis.

[0008] A second aspect of the present invention provides use of a LncRNA-ZFHX4-AS1 inhibitor in the preparation of a medicament for preventing and / or treating adrenal tumors.

[0009] It should be understood that, unless otherwise specified, in the context of the present invention, the LncRNA-ZFHX4-AS1 inhibitor refers to a substance that can specifically downregulate the expression level of LncRNA-ZFHX4-AS1, for example, antisense oligonucleotides, siRNA, shRNA, sgRNA, antagomiRs, miRNA sponges, miRNA Erasers, Target Masking and / or multi-target methods are used to downregulate the expression level and / or activity of LncRNA-ZFHX4-AS1, as long as the level and / or activity of LncRNA-ZFHX4-AS1 can be reduced.

[0010] Preferably, the LncRNA-ZFHX4-AS1 inhibitor is selected from siRNA and / or shRNA designed based on the LncRNA-ZFHX4-AS1 gene.

[0011] Preferably, the siRNA designed based on the LncRNA-ZFHX4-AS1 gene is selected from one or more of si-1, si-2, and si-3, wherein the sequence of si-1 is GGUAGAA AUGUCUAUCAUA, the sequence of si-2 is GGAUGUAACAACAUUACUA, and the sequence of si-3 is CCAGAUUCAUAAUGUUACA.

[0012] Preferably, the adrenal tumor is selected from one or more of adrenal cortical tumor, adrenal medullary tumor, adrenal stromal tumor and adrenal metastasis.

[0013] A third aspect of the present invention provides use of a reagent for detecting the expression level of LncRNA-ZFHX4-AS1 in the preparation of a product for auxiliary diagnosis and / or prognosis evaluation of adrenal tumors.

[0014] Preferably, the reagent for detecting the expression level of LncRNA-ZFHX4-AS1 includes primers for detecting the expression level of the LncRNA-ZFHX4-AS1 gene.

[0015] It should be understood that, unless otherwise specified, in the context of the present invention, the primers and / or primer pairs refer to PCR primers used to synthesize the cDNA chain of the lncRNA-ZFHX4-AS1 gene in PCR, thereby being used to detect the expression level of the lncRNA-ZFHX4-AS1 gene. Those skilled in the art are fully capable of designing corresponding primers and / or primer pairs based on the gene sequence of lncRNA-ZFHX4-AS1 using conventional methods in the field, including but not limited to molecular biology, and screening the designed primers and / or primer pairs by conventional experimental means, as long as they can achieve specific detection of lncRNA-ZFHX4-AS1 expression levels.

[0016] Preferably, the adrenal tumor is selected from one or more of adrenal cortical tumor, adrenal medullary tumor, adrenal stromal tumor and adrenal metastasis.

[0017] A fourth aspect of the present invention provides a pharmaceutical composition for preventing and / or treating adrenal tumors, comprising a LncRNA-ZFHX4-AS1 inhibitor.

[0018] Preferably, the LncRNA-ZFHX4-AS1 inhibitor is selected from siRNA and / or shRNA designed based on the LncRNA-ZFHX4-AS1 gene.

[0019] Preferably, the siRNA designed based on the LncRNA-ZFHX4-AS1 gene is selected from one or more of si-1, si-2, and si-3, wherein the sequence of si-1 is GGUAGAA AUGUCUAUCAUA, the sequence of si-2 is GGAUGUAACAACAUUACUA, and the sequence of si-3 is CCAGAUUCAUAAUGUUACA.

[0020] Preferably, the adrenal tumor is selected from one or more of adrenal cortical tumor, adrenal medullary tumor, adrenal stromal tumor and adrenal metastasis.

[0021] A fifth aspect of the present invention provides a kit for auxiliary diagnosis and / or prognosis evaluation of adrenal tumors, comprising a reagent for detecting the expression level of LncRNA-ZFHX4-AS1.

[0022] Preferably, the reagent for detecting the expression level of LncRNA-ZFHX4-AS1 includes primers for detecting the expression level of the LncRNA-ZFHX4-AS1 gene.

[0023] Preferably, the adrenal tumor is selected from one or more of adrenal cortical tumor, adrenal medullary tumor, adrenal stromal tumor and adrenal metastasis.

[0024] Preferably, the kit further comprises one or more of PCR enzyme, PCR buffer, dNTPs, and fluorescent substrate.

[0025] Preferably, the fluorescent substrate is selected from Syber Green or a fluorescently labeled probe.

[0026] ACC is a highly aggressive malignant tumor originating from the adrenal cortex. Due to the rarity of ACC and its complex and varied clinical manifestations, its early diagnosis is very difficult. According to statistics, only 10% of cases are discovered incidentally during physical examinations, which results in most patients being diagnosed in stage III or IV, with a poor overall prognosis. Prospective data indicate that the 5-year overall survival (OS) rate of patients with stage II ACC who undergo complete tumor resection is as high as 90%, while the 5-year overall survival rate of patients with advanced ACC is only about 15%. ACC patients benefit significantly from early diagnosis and treatment compared to late-stage ones, so there is an urgent need for a biomarker that can accurately predict patient prognosis.

[0027] Long noncoding RNAs (lncRNAs) are non-protein-coding RNAs with transcripts exceeding 200 nucleotides in length. With the rapid development of high-throughput sequencing technology, lncRNAs have evolved from being "transcriptional noise" to regulatory molecules with important biological functions. Studies have shown that the differential expression of various lncRNAs in human cancers is associated with the development, progression, invasion, metastasis, and drug resistance of malignant tumors. LncRNAs can also serve as molecular biomarkers, clinically used to guide diagnosis and treatment and improve patient prognosis. ZFHX4-AS1, located on chromosome 8q21.13, has been shown to be closely associated with chromosome 8q21.11 deletion syndrome. In this study, the prognostic characteristics of lncRNA-ZFHX4-AS1 in ACC patients were validated using multiple multicenter cohorts. The results showed that ACC patients with high expression of lncRNA-ZFHX4-AS1 had poorer overall survival, progression-free survival (PFI), and end-of-life (EFS) outcomes. Univariate and multivariate Cox regression analyses demonstrated that lncRNA-ZFHX4-AS1 was an independent prognostic risk factor for ACC in most cohorts. Furthermore, the nomogram constructed by the present invention is accurate and reliable, and its use in predicting patient survival is more beneficial than other clinical indicators. Subsequent in vitro experiments confirmed that LncRNA-ZFHX4-AS1 significantly promotes the proliferation, cloning, migration, and invasion of ACC cell lines. These results clearly indicate that LncRNA-ZFHX4-AS1 may serve as a pro-oncogenic factor in ACC.

[0028] Compared with the prior art, the present invention has the following technical effects:

[0029] (1) Through extensive research and screening, the present invention has found that a higher expression level of LncRNA-ZFHX4-AS1 is significantly positively correlated with adrenal tumors, which can be used to predict the malignancy and treatment prognosis of adrenal tumors, thereby screening out people with a high incidence of adrenal tumors for reasonable early prevention, significantly reducing the probability and severity of the disease, and reducing damage to human health. At the same time, it provides early assessment for patients receiving preoperative treatment and provides personalized treatment plans.

[0030] (2) This invention reveals the association between the ZFHX4-AS1 gene and adrenal tumors, suggesting that inhibiting ZFHX4-AS1 in vivo can effectively inhibit the proliferation, cloning, migration, and invasion of adrenal tumors. This has important practical significance for resolving the problems of inter-individual differences in clinical efficacy and the gaps in regression effect / prognosis assessment, and for better achieving precision treatment. It provides a new drug treatment target for humans to conquer adrenal tumors, thereby providing a new direction for subsequent drug development and clinical treatment, and has extremely high social value and market application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 Schematic diagram of Kaplan-Meier survival analysis results for different ACC cohorts.

[0032] Figure 2 Schematic diagram of ROC analysis results for different ACC cohorts.

[0033] Figure 3 Schematic diagram of the results of univariate and multivariate COX regression analysis of different ACC cohorts.

[0034] Figure 4 To construct a nomogram based on different variables to predict OS and PFI in patients with ACC.

[0035] Figure 5 Schematic diagram of the calibration results of the nomogram for estimating 1-year, 3-year, and 5-year OS and PFI.

[0036] Figure 6 The AUCs of 1-year, 3-year, and 5-year OS and PFI are shown.

[0037] Figure 7 Schematic diagram of DCA analysis results.

[0038] Figure 8 Schematic diagram of the inhibitory activity of different siRNAs on LncRNA-ZFHX4-AS1 mRNA expression in ACC cells.

[0039] Figure 9 Schematic diagram of the effect of inhibiting LncRNA-ZFHX4-AS1 expression on ACC cell proliferation.

[0040] Figure 10 Schematic diagram of the effect of inhibiting LncRNA-ZFHX4-AS1 expression on ACC cell clone formation.

[0041] Figure 11 Schematic diagram of the effect of inhibiting LncRNA-ZFHX4-AS1 expression on ACC cell migration. DETAILED DESCRIPTION

[0042] In order to make the purpose, technical solution and effect of the present invention clearer and more specific, the present invention is further described in detail with reference to the following examples. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0043] Unless otherwise specified, the cell lines listed in the context of the present invention, including H295R (human adrenocortical carcinoma cells) and SW-13 (human adrenocortical small cell carcinoma cells), were purchased from institutions such as Wuhan Punuo Sai Life Science Technology Co., Ltd. and cultured according to conventional methods in the art. All cell lines were identified by short tandem repeat analysis of the China Center for Type Culture Collection (Wuhan), and the presence of mycoplasma contamination was verified using a PCR detection kit (Shanghai Biothrive Sci), and frozen in liquid nitrogen and used for subsequent experiments. The reagents used in the present invention were all commercially available, and the LncRNA-ZFHX4-AS1 inhibitor used was a single-stranded RNA molecule designed and synthesized based on the LncRNA-ZFHX4-AS1 sequence. The experimental methods used in the present invention, such as bioinformatics analysis, nucleic acid extraction, genome sequencing, primer design, PCR, lentiviral vector construction, cell transfection, tumor cell culture, cell migration experiments, cell clone formation experiments, cell proliferation experiments, etc., are all conventional methods and techniques in the art.

[0044] Representative results from replicate experiments are presented in the accompanying figures. Data are presented as mean ± SD and mean ± SEM as indicated in the figures. All experiments were repeated at least three times. Data were analyzed using GraphPad Prism 8.0 or SPSS 22.0 software. Standard medical statistical methods, such as t-tests, Wilcoxon tests, Pearson correlation coefficients, chi-square tests, and analysis of variance, were used to compare mean values ​​between two or more groups. A p < 0.05 was considered significant.

[0045] Example 1 Confirmation of biomarkers associated with adrenal tumors

[0046] To explore the association between LncRNA-ZFHX4-AS1 expression and the prognosis of ACC patients, LncRNA-ZFHX4-AS1 expression was divided into high and low expression groups according to the optimal cutoff value, and Kaplan-Meier survival analysis was performed. Figure 1 As shown, the results showed that in the TC GA-ACC cohort, patients with high LncRNA-ZFHX4-AS1 expression had worse OS and PFI (p = 0.018, p = 0.00059). Similarly, in the GSE10927 and GSE19750 cohorts, patients in the low expression group achieved more impressive OS than those in the high expression group (*p < 0.05). In addition, in the pediatric cohort GSE76019, the high LncRNA-ZFHX4-AS1 expression group had worse EFS (p = 0.0011). The results of ROC analysis showed that in TCGA-ACC, the AUC values ​​of 1-, 3-, and 5-year OS were 0.970, 0.585, and 0.716, respectively, and the AUC values ​​of 1-, 3-, and 5-year PFI were 0.713, 0.655, and 0.764; in GSE10927, the AUC values ​​of 1-, 3-, and 5-year OS were 0.859, 0.610, and 0.603; in GSE19750, the AUC values ​​of 1-, 3-, and 5-year OS were 0.729, 0.611, and 0.628; in GSE76019, the AUC values ​​of 1-, 3-, and 5-year EFS were 0.792, 0.778, and 0.925 (see Figure 2 ). The results showed that LncRNA-ZFHX4-AS1 has a good predictive performance for the prognosis of ACC patients. Univariate and multivariate COX regression analysis showed that, except for the GSE76019 cohort, ZFHX4-AS1 expression was an independent risk factor for OS and PF I (*p<0.05) ( Figure 3 ).

[0047] In recent years, nomograms have been widely used as a risk assessment tool for the prognosis of cancer patients. To improve the clinical applicability of this study, a nomogram was established to predict the 1-year, 3-year, and 5-year OS and PFI of ACC patients based on the scores of predictive variables (including LncRNA-ZFHX4-AS1, age, T stage, N stage, and gender). The results are shown in Figure 2. Figure 4 The calibration curve shows that the 1-year, 3-year, and 5-year OS and PFI estimated by the nomogram are consistent with the actual situation (see Figure 5 In addition, the AUC values ​​of the nomogram for 1-year, 3-year, and 5-year OS were 0.939, 0.751, and 0.905, respectively, and the AUC values ​​of the nomogram for 1-year, 3-year, and 5-year PFI were 0.744, 0.763, and 0.855, respectively (see Figure 6), which shows that the constructed nomogram is accurate and reliable. Decision Curve Analyses (DCA) can estimate the net benefit of the model based on the difference between the number of true positive results and false positive results, and is widely used to evaluate whether the nomogram is effective in assisting decision making. The DCA results suggest that the use of nomograms to predict the 1-, 3-, and 5-year survival rates of ACC patients has a greater net benefit and is more effective than other clinical indicators (see Figure 7 In conclusion, LncRNA-ZFHX4-AS1 is a reliable new biomarker that can accurately predict the prognosis of ACC patients.

[0048] Example 2 Effects of LncRNA-ZFHX4-AS1 on the Behavior and Function of Adrenal Tumor Cells

[0049] To evaluate whether the expression of LncRNA-ZFHX4-AS1 might affect the proliferation of ACC cells, three siRNAs targeting LncRNA-ZFHX4-AS1 were designed (si-1: sequence GGUAGAAAUGUCUAUCAUA (5'-3'), si-2: sequence GGAUGUAACAACAUU ACUA (5'-3'), si-3: sequence CCAGAUUCAUAAUGUUACA (5'-3')). The siRNAs were transfected into SW-13 and H295R cells, respectively. The specific steps are as follows:

[0050] (1) One day before transfection, cells were seeded in a 6-well plate. 100 pmol of siRNA was added to 200 μL of serum-free culture medium and gently mixed.

[0051] (2) Dilute 5 μL RNAi-Mate with 200 μL serum-free medium, mix gently, and let stand at room temperature for 5 minutes.

[0052] (3) Mix the diluted siRNA and RNAi-Mate reagent, mix gently, and let it stand at room temperature for 10 minutes to form the siRNA / RNAi-Mate complex.

[0053] (4) Add 400 μL of siRNA / RNAi-Mate complex to a cell culture plate containing 1.6 mL of culture medium, gently shake the cell culture plate back and forth, and place it in a 37°C, 5% CO2 incubator for 24 h. After that, RNA was extracted and the expression level of LncRNA-ZFHX4-AS1 mRNA in the cells was detected by real-time fluorescence quantitative PCR (RT-qPCR).

[0054] Test results such as Figure 8The results showed that in H295R cells, the three siRNA sequences all caused different degrees of reduction in the expression level of LncRNA-ZFHX4-AS1 mRNA. si-1, si-2, and si-3 showed statistical differences compared with the si-NC group (**p < 0.01), with si-1 and si-3 having higher inhibitory efficiency. In SW-13 cells, the three siRNA sequences all caused different degrees of reduction in the expression level of LncRNA-ZFHX4-AS1 mRNA. si-1, si-2, and si-3 showed statistical differences compared with the si-NC group (**p < 0.01), with si-1 and si-3 having higher inhibitory efficiency.

[0055] After confirming that the designed siRNA could inhibit the expression of LNCRNA-ZFHX4-AS1 by targeting it, the above siRNA was used to study its effect on the proliferation of adrenal tumor cells. The specific steps are as follows:

[0056] (1) siRNAs targeting LNCRNA-ZFHX4-AS1 (si-1 and si-3) were transfected into SW-13 and H295R cells, respectively.

[0057] (2) When the cells grow to the logarithmic phase, trypsinize and count them, and select the appropriate cell density according to the doubling time of various cells (SW-13: 1×10 4 cells / mL; H295R: 5×10 4 Cells were seeded into 96-well plates (3 replicates) and 100 μL of cell suspension was added to each well.

[0058] (3) Culture the cells in a 37°C incubator and collect the cells at 24 h, 48 h, 72 h, and 96 h. Add 10 μL of CCK-8 to each well and incubate the culture plate in the incubator for 2 h. Measure the absorbance at 450 nm to evaluate the cell proliferation.

[0059] The results are as follows Figure 9 The results showed that after silencing the LncRNA-ZFHX4-AS 1 gene using siRNA, the proliferation ability of adrenal cancer cells was significantly weakened, and the growth of adrenal cancer cells was significantly inhibited. In other words, knockdown of LncRNA-ZFHX4-AS1 significantly reduced the proliferation activity of adrenal cancer cells, and the difference was statistically significant compared with the control group (***p < 0.0001).

[0060] Subsequently, we used siRNA targeting LncRNA-ZFHX4-AS1 to study its effect on adrenal tumor cell clone formation. The specific steps are as follows:

[0061] (1) siRNAs targeting LNCRNA-ZFHX4-AS1 (si-1 and si-3) were transfected into SW-13 and H295R cells, respectively.

[0062] (2) When the cells grow to the logarithmic phase, trypsinize and count them, and select the appropriate cell density according to the doubling time of various cells (SW-13: 1×10 4 cells / mL; H295R: 5×10 4 The cells were inoculated into a dish containing 10 mL of 37°C pre-warmed culture medium, and gently rotated to disperse the cells evenly. The dish was then cultured in a cell culture incubator at 37°C with 5% CO2 and saturated humidity.

[0063] (3) When visible clones appear in the culture dish (approximately 10-14 days), terminate the culture, discard the supernatant, and carefully rinse twice with PBS. Add 500 μL of 4% paraformaldehyde to each well and fix for 10 minutes, then discard it. Then add 1 mL of methanol containing 0.5% crystal violet and stain for 30 minutes. Discard the methanol and wash the residual methanol 2-3 times with water. Cell clones can be observed. Under a microscope, a valid clone is counted when the number of cells is greater than 50.

[0064] Test results such as Figure 10 The results showed that in H295R cells, the clone-forming ability of cells transfected with si-1 and si-3 was significantly inhibited compared with the control group (***p < 0.001); in SW-13 cells, the clone-forming ability of cells transfected with si-1 and si-3 was significantly decreased compared with the control group (****p < 0.0001). In summary, knockdown of LncRNA-ZFHX4-AS1 significantly inhibited the clone-forming ability of ACC cells.

[0065] Furthermore, we used siRNA targeting LncRNA-ZFHX4-AS1 to study its effect on adrenal tumor cell migration using Corning Transwell Migration 8μm pore chambers and 24-well plates. The specific steps are as follows:

[0066] (1) Place the chamber in a 24-well plate and add 1 mL of basal culture medium to each well to moisten it.

[0067] (2) After trypsin digestion of cells in the logarithmic growth phase, resuspend them in basal medium to form a cell suspension, count them, and adjust the cell density to 5×10 5 pieces / mL.

[0068] (3) The basal culture medium in the chamber and the 24-well plate was aspirated and discarded, 200 μL of cell suspension was added to the upper chamber of the transwell chamber, and 600 μL of complete culture medium (basal culture medium + 10% fetal bovine serum) was added to the lower chamber of the 24-well culture plate.

[0069] (4) Place the culture plate in a CO2 incubator at 37°C and continue incubation for 14 hours.

[0070] (5) The chamber was removed, rinsed twice with PBS, fixed with 4% paraformaldehyde in a 24-well plate for 20 min, and stained with crystal violet solution for 15 min.

[0071] (6) Carefully wipe off the cells in the upper layer of the microporous membrane of the chamber with a cotton swab and take pictures under an inverted microscope.

[0072] Test results such as Figure 11 As shown. The results showed that, in both H295R and SW-13 cells, inhibition of intracellular LncRNA-ZFHX4-AS1 expression using siRNA inhibitors significantly inhibited the migration of adrenal tumor cells compared to the blank vector siNC group. This difference was statistically significant (*p < 0.05, ****p < 0.0001). The above results clearly indicate that LncRNA-ZFHX4-AS1 is a gene that is closely related to the development and progression of adrenal tumors. Compared with normal adrenal cells or tissues, LncRNA-ZFHX4-AS1 is significantly overexpressed in adrenal tumor cells, indicating that LncRNA-ZFH X4-AS1 can be used for the early diagnosis and prognosis of adrenal tumors. Inhibition of LncRNA-ZFHX4-AS1 expression can significantly inhibit the proliferation, cloning, migration, and invasion of adrenal tumor cells. At the same time, by revealing the correlation between the LncRNA-ZFHX4-AS1 gene and the development and progression of adrenal tumors, this study has important practical significance for addressing the gap in clinical efficacy and prognostic assessment for adrenal tumors and better implementing precision treatment. It clearly demonstrates that inhibiting LncRNA-ZFHX4-AS1 expression can significantly inhibit adrenal tumor cells, thereby significantly suppressing the development, progression, and metastasis of adrenal tumors. LncRNA-ZFHX4-AS1 can serve as a new target for non-invasive diagnosis, prognosis, and precision targeted therapy of adrenal tumors. This provides a new drug therapeutic target for the fight against adrenal tumors, thereby offering a new direction for subsequent drug development and clinical treatment, and has extremely high social value and market application prospects.

[0073] The above detailed description of the analytical methods involved in the present invention provides a detailed introduction. It should be noted that the above description is intended solely to help those skilled in the art better understand the methods and concepts of the present invention, and is not intended to limit the relevant content. Without departing from the principles of the present invention, those skilled in the art may make appropriate adjustments or modifications to the present invention, and such adjustments and modifications shall also fall within the scope of protection of the present invention.

Claims

1. Use of a lncRNA-ZFHX4-AS1 inhibitor in the preparation of a medicament for preventing and / or treating adrenal tumors; characterized in that: The LncRNA-ZFHX4-AS1 inhibitor is selected from siRNA and / or shRNA designed based on the LncRNA-ZFHX4-AS1 gene; the adrenal tumor is selected from one or more of adrenocortical tumor, adrenal medullary tumor, adrenal stromal tumor, and adrenal metastasis.

2. Use of a reagent for detecting the expression level of LncRNA-ZFHX4-AS1 in the preparation of a product for auxiliary diagnosis and / or prognosis evaluation of adrenal tumors, characterized in that: The adrenal tumor is selected from one or more of adrenal cortical tumor, adrenal medullary tumor, adrenal stromal tumor and adrenal metastatic tumor.

3. The use according to claim 2, characterized in that The reagent for detecting the expression level of LncRNA-ZFHX4-AS1 includes primers for detecting the expression level of the LncRNA-ZFHX4-AS1 gene.

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