Application of Rad17 as a marker and therapeutic target for cervical cancer

Abstract

The present invention belongs to the field of biomedicine technology, and discloses the application of Rad17 as a cervical cancer marker and therapeutic target. Through research, the present invention provides a new cervical cancer diagnostic marker Rad17, which can clearly characterize the occurrence and development of cervical cancer. The marker shows a specific high expression phenomenon in the cervical cancer tissue of clinical patients compared with the corresponding normal cervical tissue, and also shows high expression in human cervical cancer cell lines; and it is confirmed that knocking down Rad17 will inhibit the malignant characteristics of cervical cancer. As a diagnostic marker for cervical cancer, Rad17 provides a new molecular target for the treatment of cervical cancer and provides a new idea for the treatment of cervical cancer.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to the application of Rad17 as a cervical cancer marker and therapeutic target. Background Art

[0002] Globally, cervical cancer is the fourth most common cancer in terms of morbidity and mortality among female cancers, second only to breast cancer, colorectal cancer, and lung cancer. Persistent high-risk HPV infection is the main factor in the occurrence of cervical cancer, but its mechanism is not clear. The current HPV vaccine can only prevent several types of HPV infection, and has no therapeutic effect on patients who are already infected. How to reduce the incidence of cervical cancer remains an important issue that needs to be addressed. Therefore, in-depth research on the molecular mechanisms of the occurrence and development of cervical cancer is of great significance for the development of new diagnostic markers and drug targets.

[0003] Pap smear is currently a clinical way to detect cervical precancerous lesions and early cervical cancer. This method performs preliminary screening of cervical exfoliated cells with good specificity but poor sensitivity. HPV testing can assist in diagnosis, but HPV infection is only a high-risk factor and cannot determine whether it has developed into cancer. Colposcopy can further examine suspected cervical cancer, but this method has high professional requirements for operators and is difficult to popularize in areas of my country with backward medical levels. Therefore, it is particularly important to find new tumor diagnostic molecular markers and therapeutic targets.

[0004] Cell cycle checkpoints are some control points in the cell cycle process. When DNA is damaged (sometimes also when cells encounter adverse conditions such as nutrient deficiency), these control points slow down or temporarily stop the cell cycle process to provide cells with enough time to repair damaged DNA (or survive the crisis). The cell cycle checkpoint-related protein Rad17 is a key control protein in the process of cell response to DNA damage and replication fork block signal transduction, and plays a very important role in DNA damage and DNA replication control. There is no report on the correlation between Rad17 and cervical cancer. Summary of the invention

[0005] The purpose of the present invention is to address the above problems and provide a new application of Rad17 as a cervical cancer marker: application of a substance for detecting the expression level of a biomarker in the preparation of a product for diagnosing cervical cancer, wherein the biomarker is Rad17.

[0006] In the above application technology scheme, the Rad17 gene or protein expression level in the cervical tissue of an individual suffering from or suspected of suffering from cervical cancer is detected.

[0007] In the above-mentioned application technology scheme, if the Rad17 gene or protein expression level in the individual's diseased cervical tissue shows a specific high expression compared with the normal cervical tissue, it indicates that the individual is a high-risk patient for cervical cancer.

[0008] In the above application technology solution, the product is a reagent or a kit.

[0009] In the above application technology scheme, the substance is used to detect the protein expression level of Rad17 gene or the mRNA expression level of Rad17 gene.

[0010] The present invention also provides the use of Rad17 as a target in screening / preparing drugs for treating cervical cancer, inhibiting the expression of Rad17 gene and suppressing the malignant characteristics of cervical cancer.

[0011] In the above application technology scheme, the expression of Rad17 is inhibited, thereby inhibiting the expression of HMGCS1 in cervical cells, thereby reducing total cholesterol and inhibiting the proliferation, migration and invasion of cervical cancer cells.

[0012] The present invention also provides the use of a reagent for inhibiting the expression of Rad17 in preparing a medicine for treating cervical cancer.

[0013] In the above application technology scheme, the agent that inhibits the expression of Rad17 inhibits the expression of Rad17, thereby inhibiting the expression of HMGCS1 in cervical cells, thereby reducing total cholesterol and inhibiting the proliferation, migration and invasion of cervical cancer cells.

[0014] In the above application technology solution, the reagent for inhibiting the expression of Rad17 includes a nucleic acid molecule, a protein molecule or a compound; preferably shRNA.

[0015] The beneficial effects of the present invention are: providing a new cervical cancer diagnostic marker Rad17, which can clearly characterize the occurrence and development of cervical cancer. The marker shows a specific high expression phenomenon in human clinical cervical cancer tissues compared with corresponding normal cervical tissues; it also shows high expression in human cervical cancer cell lines; and it is confirmed that knocking down Rad17 will inhibit the malignant characteristics of cervical cancer. Rad17, as a diagnostic marker for cervical cancer, provides a new molecular target for the treatment of cervical cancer and provides a new idea for the treatment of cervical cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 The aim of this study was to analyze the expression of Rad17 in normal human cervical tissue and human cervical cancer tissue in the GEO database.

[0017] Figure 2 The GEPIA online database was used to analyze the effect of Rad17 on the disease-free survival of patients with cervical cancer.

[0018] Figure 3 RT-qPCR was used to detect the mRNA level of Rad17 in normal human cervical tissue and human cervical cancer tissue.

[0019] Figure 4 IHC is used to detect the protein expression level of Rad17 in human normal cervical cancer tissue, CIN tissue, and cancer tissue.

[0020] Figure 5 Western blot was used to detect the protein expression level of Rad17 in immortalized human normal cervical epithelial cells (H8) and several human cervical cancer cell lines (CaSki, SiHa and HeLa).

[0021] Figure 6 RT-qPCR was used to detect the relative mRNA expression of Rad17 in immortalized human normal cervical epithelial cells (H8) and several human cervical cancer cell lines (CaSki, SiHa and HeLa).

[0022] Figure 7 Western blot and RT-qPCR were used to verify the knockdown efficiency of Rad17 in CaSki and SiHa.

[0023] Figure 8 RTCA was used to detect the proliferation ability of CaSki and SiHa after knockdown of Rad17.

[0024] Fig. 9 CCK8 was used to detect the proliferation ability after knocking down Rad17 in CaSki and SiHa.

[0025] Fig.10 The clone formation assay was used to detect the proliferation ability after knocking down Rad17 in CaSki and SiHa.

[0026] Fig.11 The scratch assay was used to detect the migration ability of CaSki and SiHa after knockdown of Rad17.

[0027] Fig.12 The Transwell assay was used to detect the migration and invasion abilities of CaSki and SiHa after knocking down Rad17.

[0028] Fig.13 The GEPIA database was used to analyze the correlation between HMGCS1 and Rad17 in cervical cancer tissues.

[0029] Fig.14 Western blot and RT-qPCR were used to detect the protein and mRNA levels of HMGCS1 after knockdown of Rad17 in CaSki.

[0030] Fig.15 The total cholesterol quantification experiment was used to detect the total cholesterol content after knocking down Rad17 in CaSki.

[0031] Fig.16 The total cholesterol quantification experiment was used to detect the total cholesterol content after overexpression of Rad17 in CaSki and after overexpressing Rad17 cells were treated with HMGCS1 inhibitor Dipyridamole.

[0032] Fig.17 The GEO database was used to analyze the expression of HMGCS1 in cervical cancer.

[0033] Fig.18 Western blot and RT-qPCR were used to detect the protein and mRNA expression levels of HMGCS1 in immortalized human normal cervical epithelial cells and several human cervical cancer cell lines.

[0034] Fig.19 The knockdown effect of HMGCS1 was detected by western blot.

[0035] Fig. 20 The total cholesterol quantification experiment was used to detect the total cholesterol content after knocking down HMGCS1 in CaSki.

[0036] Fig.21 CCK8 was used to detect the proliferation ability after knocking down HMGCS1 in CaSki and SiHa.

[0037] Fig. 22 The Transwell assay was used to detect the migration and invasion abilities of CaSki and SiHa cells after knocking down HMGCS1.

[0038] Fig.23 It is the comparison of the volume of tumor tissue with downregulated Rad17 and that of control tumor tissue.

[0039] Fig.24 The figure is the comparison of the weight of tumor tissue with downregulated Rad17 and that of control tumor tissue.

[0040] Fig.25 This is the change of HMGCS1 detected by IHC on day 15 of the transplanted tumor experiment. DETAILED DESCRIPTION

[0041] The present invention will be further described below in conjunction with embodiments, but the present invention is not limited thereto.

[0042] The experimental methods in the following examples are conventional methods unless otherwise specified.

[0043] Example 1

[0044] 1 Experimental Materials and Methods

[0045] 1.1 Cell culture

[0046] This experiment used three human cervical cancer cell lines, CaSki, SiHa and HeLa cells, and one immortalized normal human cervical epithelial cell line, H8. CaSki cell line (Procell, Wuhan) was cultured in RPMI-1640 medium (C11875500BT, Gibco) containing 10% fetal bovine serum (C8010, Adamas, USA); SiHa cells (Procell, Wuhan) were cultured in MEM-α medium (L570KJ, Basalmedia) supplemented with 10% fetal bovine serum; HeLa cells (Procell, Wuhan) were cultured in DMEM medium (C11995500BT, Gibco) containing 10% fetal bovine serum; H8 cells (Qingqi, Shanghai) were cultured in DMEM medium supplemented with 10% fetal bovine serum. All these cell lines were cultured at 37°C and 5% CO 2 Cultivated in a standard environment.

[0047] 1.2 Lentiviral packaging and construction of cells with knockdown of Rad17 or HMGCS1

[0048] In order to obtain cells with knockdown of Rad17 expression, shRNA virus targeting Rad17 was prepared by lentiviral packaging. The process was as follows: using a lentiviral packaging three-plasmid system, the lentiviral packaging plasmids pMD2G, psPAX2, and shRad17 (Sigma-Aldrich, USA) or pLKO.1-TRC vector (shRad17 control (NC) plasmid, Sigma-Aldrich, USA) were packaged with lentivirus at a mass ratio of 2:9:9. When HEK 293T cells were 70% confluent, they were transfected into cells through PEI (24765-1, Polysciences) according to the instructions of the plasmid system product manual. The viral supernatant was collected after 48 hours. When the cells reached 50% confluence, the shRNA virus targeting Rad17 was transfected into CaSki and SiHa cells, and infection was mediated by polybrene reagent (C0351, Beyotime) according to the product instructions. Puromycin (ST551, Beyotime) was used for selection after 48 hours. The construction of HMGCS1 knockdown cells was the same as above, and the shHMGCS1 sequence is shown in Table 1.

[0049] Table 1 shRNA sequences

[0050]

[0051] 1.3 CCK-8 experiment

[0052] The cells were plated at 3 × 10 3 Cells were inoculated into 96-well plates at a density of 10 cells / well, with 5 replicate wells per group. Subsequently, 10 μL of CCK-8 solution was added to each well at 0, 24, 48, 72, and 96 h, respectively, and incubated at 37 °C for 1.5 h. The absorbance at a wavelength of 450 nm was measured using an ELISA reader. Cell viability curves were drawn based on the absorbance values.

[0053] 1.4 Clone formation assay

[0054] The cells were plated at 3 × 10 3 The cells were seeded into a six-well plate at a density of 1 / well and cultured for 14 days. Next, the cells were fixed with 4% paraformaldehyde and stained with 0.1% crystal violet. The cells were photographed with a mobile phone and counted using Image J software to calculate the clone formation rate.

[0055] 1.5 Immunohistochemistry (IHC)

[0056] Paraffin sections were subjected to IHC staining. Paraffin sections were dewaxed in xylene and a series of different concentrations of ethanol. After antigen retrieval, blocking of endogenous peroxidase, and blocking, they were incubated with Rad17 antibody (#13358-1-AP, 1:200 Proteintech, USA) at 4°C overnight. Enzyme-labeled goat anti-mouse / rabbit IgG polymer secondary antibody (PV-6000, zsbio, China) was incubated at room temperature for 20 minutes. Then, DAB color development, hematoxylin staining, graded ethanol dehydration, and neutral resin sealing were performed. 1.6 Real-time cell analysis (RTCA)

[0057] For RTCA, cells were collected at 3 × 10 3 The density of cells / well was seeded in E-Plate Insert 16 (6465382001, Agilent, USA), and the real-time cell index was measured by xCELLigence RTCA S16.

[0058] 1.7 Real-time quantitative polymerase chain reaction (RT-qPCR).

[0059] Total RNA from cervical cancer cell lines was extracted using RNAiso Plus (9109, Takara, Beijing) and reverse transcribed into cDNA template using ABScript III RTMaster Mix for qPCR with gDNARemover (RK20429, ABclonal, Wuhan). The NCBI Gene ID of Rad17: 5884, the NCBI Gene ID of HMGCS1: 3157, and the designed specific primer sequences (shown in Table 2) were synthesized by Tsingke (China). Subsequently, the cDNA template, specific primers, and SYBR Green Fast qPCR Mix reagent (RK21203, ABclonal, China) were mixed to amplify the target gene, and the RT-qPCR reaction was performed on the Roche LightCycler480II system. The expression of Rad17 or HMGCS1 was normalized with GAPDH or β-actin. The relative expression was calculated using the 2-ΔΔCT method.

[0060] Table 2 RT-qPCR primer sequences

[0061]

[0062] The cDNA obtained by reverse transcription was used as a template, 3 replicate wells were set up for each group of samples, and the reaction system was prepared on ice according to the components listed below.

[0063]

[0064] Perform qPCR reaction as follows:

[0065]

[0066] Calculation results: The relative mRNA expression of the target gene was calculated according to the Ct value of the target gene obtained by qPCR reaction and the Ct value of the internal reference gene according to 2-ΔΔCt.

[0067] 1.8 Scratch test

[0068] Scratch assay was used to evaluate cell migration ability. Cells were treated in serum-free medium for 12 h. Scratch was formed by a 10 μl pipette tip and recorded by an inverted fluorescence microscope (DMi8, Leica, Germany). After 24 h, the cell migration distance was measured using Image J software and the cell migration ability was calculated.

[0069] 1.9 Transwell experiment

[0070] Transwell assay was used to evaluate the migration and invasion ability of cells. 4 cells / well) were seeded in the upper chamber of Transwell (14341, labselect, Anhui). For invasion assay, cell suspension (7.5×10 4 Cells / well) were seeded in the upper chamber of Transwell, and the matrix gel (356234, BD Biosciences, San Jose, USA) was diluted 1:9 with serum-free medium and pre-coated on the upper chamber 1 hour in advance. 600ul of medium containing 20% ​​fetal bovine serum was added to the lower chamber. After incubation for 48 hours, the cells in the upper chamber were taken and washed twice with PBS. Then the cells were fixed with 4% paraformaldehyde and stained with 0.1% crystal violet (G1014, Servicebio, Wuhan). The number of cells in the lower chamber was observed under an upright microscope, and 3 fields of view were randomly selected in each image for counting.

[0071] 1.10 Quantitative detection of total cholesterol

[0072] Quantitative detection of total cellular cholesterol was performed according to the instructions of the total cholesterol (T-CHO) test kit (A111-1-1, Nanjing Jiancheng, China).

[0073] 1.11 Immunoblotting

[0074] Total protein of cervical cancer cell lines was isolated using RIPA lysis buffer containing PMSF (R0010, Solarbio, China) and quantified using a BCA assay kit (P0010, beyotime, China). Equal amounts of protein were then loaded on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gels and transferred to PVDF membranes (ISEQ00010, Millipore, China). After blocking with 5% skim milk at room temperature for 1 h, the membranes were incubated overnight at 4°C with one of the following specific antibodies: anti-Rad17 antibody (1:6,000, 13358-1-AP; Proteintech, USA), GAPDH (1:10000; 60004-1-Ig, Proteintech). Subsequently, HRP-conjugated goat anti-rabbit antibody (1:10000; 5220-0336, Seracare, USA) or HRP-labeled goat anti-mouse antibody (1:10000; 5220-0341, Seracare) was incubated with the membrane at room temperature for 1 h. Protein images were collected using an ECL kit (AP34L024 / AP34L025, life-ilab, China) and an integrated imager (ImageQuant LAS 500, GE Healthcare Life Sciences), and quantified using Image J software, using β-actin or GAPDH as an internal reference.

[0075] 1.12 Xenograft Tumor Model Analysis

[0076] This study was approved by the Ethics Committee of Chongqing Medical University. All animal experimental procedures were performed in accordance with the Guide for the Care and Use of Laboratory Animals. Balb / c nude mice (4–5 weeks, 18–20 g) were purchased from Hunan Slake Jingda Laboratory Animal Co., Ltd., China, and housed in standard conditions. sh-Rad17 or sh-NC (control) cells were stably transfected with puro antibiotics (ST551, beyotime). Then, 7 × 10 cells were added to each animal. 6 SiHa cells stably transfected with sh-Rad17 or sh-NC were subcutaneously injected into the left side of the back at a density of 10 cells / mL. 3 )=(length×width 2 The tumor volume was measured every 3 to 5 days using the formula ) / 2. The mice were killed after 15 days and the tumor weight was measured.

[0077] 1.13 Statistical analysis

[0078] Data are expressed as mean ± SD and analyzed using GraphPad Prism software (San Diego, CA, USA). Comparisons were made using t-tests or one-way ANOVA followed by Bonferroni test, with p < 0.05.

[0079] 2 Results

[0080] 2.1 Abnormal upregulation of Rad17 expression in cervical cancer

[0081] We first detected the expression of Rad17 in cervical cancer and its effect on the disease-free survival of cervical cancer patients. We analyzed the expression of Rad17 in 14 normal cervical tissues and 14 HPV16-positive cervical cancer tissues in the GEO database. Figure 1 The results showed that the expression of Rad17 in HPV16-positive cervical cancer tissues was significantly higher than that in normal tissues. The GPIA database survival analysis showed that ( Figure 2 ), patients with cervical cancer with high Rad17 expression had a shorter disease-free survival than those with low Rad17 expression. This suggests that Rad17 may affect the occurrence and development of cervical cancer. In addition, we detected the Rad17 mRNA levels in 11 normal cervical tissues and 18 cancer tissues by quantitative reverse transcription polymerase chain reaction (RT-qPCR) analysis. Figure 3 The results showed that the expression of Rad17 was significantly increased in cervical cancer tissues. To verify the expression of Rad17 at the protein level, an additional 15 human cervical tissue samples were subjected to immunohistochemistry (IHC) analysis using Rad17-specific antibodies, including 5 normal, 5 precancerous lesions, and 5 cervical cancer tissue samples. The results showed that the staining intensity of Rad17 in cancer tissues was much higher than that in normal tissues, indicating that Rad17 was significantly upregulated in cervical cancer tissues ( Figure 4 ). The expression of Rad17 was then detected in several cervical cancer cell lines (such as CaSki, SiHa, and HeLa) by RT-qPCR and western blot. Compared with the immortalized normal cervical epithelial cell line (H8), cervical cancer cells showed higher Rad17 mRNA and protein levels ( Figure 5 and 6 ), which is consistent with our observations at the organizational level.

[0082] 2.2 Knockdown of Rad17 inhibits proliferation, migration and invasion of cervical cancer cells

[0083] To examine whether Rad17 regulates the development of cervical cancer, Rad17 was knocked down using shRNA and the effect on cell proliferation was examined. Knockdown cells were generated from CaSki or SiHa cell lines that were transduced with two different lentiviruses expressing Rad17-shRNA and stably selected by puromycin. Figure 7Western blot or RT-qPCR analysis in the generated cells showed that the knockdown efficiency was high. Real-time cell analysis (RTCA) was used to determine the effect of Rad17 on cell growth. Our results showed that the growth rate of the knockdown group was significantly lower than that of the control group ( Figure 8 ). Similarly, CCK-8 analysis showed that Rad17 silencing significantly inhibited cell proliferation of cervical cancer cells ( Fig. 9 ). Similarly, the number of colonies in the Rad17 knockdown group was less than that in the control group ( Fig.10 ).

[0084] In addition to cell proliferation, we also tested the role of Rad17 in cervical cancer cell migration and invasion. Using a scratch healing assay, knockdown cells were compared with control cells and the results showed that knockdown of Rad17 significantly reduced the wound healing rate ( Fig.11 ).

[0085] Similarly, Transwell migration and invasion assays were used to detect the effects of Rad17 knockdown on cell migration and invasion abilities. Fig.12 The results showed that fewer cells migrated in the Rad17 knockout group compared with the control group. In addition, the number of cells that invaded was significantly reduced after knocking down Rad17 ( Fig.12 ). Taken together, these results suggest that Rad17 promotes the proliferation, migration, and invasion of cervical cancer cells.

[0086] 2.3 Knockdown of Rad17 inhibits the expression of HMGCS1 in cervical cancer cells

[0087] like Fig.13 The results showed that GEPIA database analysis showed that HMGCS1 was highly correlated with Rad17 in cervical cancer tissues, so we speculated that Rad17 affects the occurrence and development of cervical cancer by regulating HMGCS1. RT-qPCR and western blot experiments confirmed that knocking down Rad17 reduced the mRNA and protein levels of HMGCS1 in CaSki cells. Fig.14 Since HMGCS1 is one of the key enzymes in the cholesterol biosynthesis pathway and catalyzes the conversion of acetoacetyl-CoA to HMG-CoA, we investigated whether Rad17 regulates cholesterol biosynthesis by regulating HMGCS1. Fig.15 Silencing Rad17 downregulated the total cholesterol content in CaSki cells. In addition, Dipyridamole, an indirect inhibitor of HMGCS1, partially offset the increase in total cholesterol content induced by Rad17. Fig.16 Taken together, these data suggest that Rad17 affects the development and progression of cervical cancer by regulating 3-hydroxy-3-methylglutaryl-CoA synthase 1 (HMGCS1).

[0088] 2.4 Silencing HMGCS1 reduces total cholesterol and inhibits the proliferation and migration of cervical cancer cells

[0089] The link between HMGCS1 and cholesterol metabolism has been widely recognized. However, whether HMGCS1 causes cervical cancer is still unclear. Recent studies have shown that HMGCS1 is closely related to cancer. Therefore, we intend to explore whether HMGCS1 affects cervical cancer. The GEO database shows that HMGCS1 is highly expressed in cervical cancer tissues. Fig.17 Western blot and RT-qPCR showed that the expression levels of HMGCS1 protein and mRNA in cervical cancer cells (CaSki, SiHa and HeLa) were significantly higher than those in normal cervical epithelial cells (H8). Fig.18 We generated stable HMGCS1 knockdown cervical cancer cells. Western blot analysis confirmed the efficiency of lentiviral infection of HMGCS1. Fig.19 We first tested the effect of HMGCS1 on cholesterol metabolism. The results showed that knocking down HMGCS1 reduced the total cholesterol content in CaSki cells. Fig. 20 .

[0090] In order to verify whether HMGCS1 affects the growth of cervical cancer, CCK8 experiments were performed. The results showed that silencing HMGCS1 significantly inhibited the growth ability of cervical cells. Fig.21 Transwell migration and invasion assays showed that knocking down HMGCS1 inhibited the migration and invasion abilities of cervical cancer cells. Fig. 22 .

[0091] 2.5 Knockdown of Rad17 inhibits xenograft tumor formation

[0092] To test the effect of Rad17 on tumor growth in vivo, SiHa cells were stably transfected with shRad17 and injected subcutaneously into the left back of nude mice. Fig.23 and Fig.24 As shown, tumor size and weight can be suppressed by downregulating Rad17. Fig.25 As shown, knocking down Rad17 significantly inhibited the expression of HMGCS1 protein level. In conclusion, Rad17 also affects cholesterol synthesis in vitro by affecting HMGCS1, thereby affecting the growth of cervical cancer cell transplants.

[0093] 3 Conclusion

[0094] The present study shows that: 1) Rad17 expression is increased in cervical cancer tissues and is associated with poor prognosis. 2) Specific knockdown of Rad17 inhibits the malignant progression of cervical cancer in vitro and in vivo. 3) Rad17 regulates cholesterol synthesis by regulating the expression of HMGCS1, thereby affecting the malignant progression of cervical cancer.

Claims

1. Use of a substance for detecting the expression level of a biomarker in the preparation of a product for diagnosing cervical cancer, characterized in that: The biomarker is Rad17.

2. The use according to claim 1, characterized in that: Detection of Rad17 gene or protein expression levels in cervical tissue of individuals suffering from or suspected of suffering from cervical cancer.

3. The use according to claim 1, characterized in that: If the Rad17 gene or protein expression level in the individual's diseased cervical tissue shows a specific high expression compared with that in the normal cervical tissue, it indicates that the individual is a high-risk patient for cervical cancer.

4. The use according to claim 1, characterized in that: The product is a reagent or a kit.

5. The use according to claim 1, characterized in that: The substance is used to detect the protein expression level of the Rad17 gene or the mRNA expression level of the Rad17 gene.

6. Application of Rad17 as a target in screening drugs for treating cervical cancer, characterized in that: Inhibit the expression of Rad17 gene and suppress the malignant characteristics of cervical cancer.

7. The use according to claim 6, characterized in that: Inhibiting the expression of Rad17, thereby inhibiting the expression of HMGCS1 in cervical cells, reduces total cholesterol and inhibits the proliferation, migration and invasion of cervical cancer cells.

Images