Effects Of Rage On Proliferation,migration,apoptosis And Sensitivity To Cisplatin Chemotherapy Of Cervical Cancer Cells

Cervical cancer is one of the most common female malignant tumors,and it seriously threatens women’s health.The mechanism of tumor occurrence and development is very complicated,and gene mutations are one of the important reasons for the malignant transformation of normal tissue cells.The receptor for advanced glycation end products（RAGE）is highly expressed in various malignant tumor tissues and is closely associated with poor clinical prognosis.In addition,it has been reported that RAGE is also related to chemotherapy resistance of tumor cells.However,RAGE has not been thoroughly studied in cervical cancer.In this study,small molecule RNA interference technology was used to down-regulate the expression of RAGE.To explore the relationship between RAGE and proliferation,migration and apoptosis of cervical cancer.On this basis,further explore the effect of RAGE on the sensitivity of cervical cancer to cisplatin chemotherapy and its possible mechanism.PART Ⅰ Effects of RAGE on proliferation,migration and apoptosis of cervical cancer cellsObjective: To investigate the effects of RAGE on proliferation,migration and apoptosis of human cervical cancer cell lines.Methods: Real-time fluorescence quantitative PCR（qPCR）was used to detect the expression level of RAGE in three common human cervical cancer cell lines.One si-NC and three si-RAGE small interference sequences were designed,and the best small interference sequences were selected by q PCR and Western blot.The cervical cancer cells were transfected with the selected small interference.The proliferation ability of the cells in the si-NC group and the si-RAGE group was detected by CCK8,the migration ability of the cells in the si-NC group and the si-RAGE group was detected by scratch test,and the apoptosis rate of the cells in the si-NC group and the si-RAGE group was detected by flow cytometry.Results: The results of q PCR showed that the expression of RAGE was the highest in SIHA cells,followed by He La cells.The results of q PCR and Western blot showed that si-RAGE2 had the best knockout efficiency.CCK8 results showed that down-regulation of RAGE expression significantly reduced the proliferation ability of Si Ha and He La cells（P<0.001,P<0.001）.Cell scratch results showed that down-regulation of RAGE expression reduced the migration ability of Si Ha and He La cells.（P<0.05,P<0.05）.Flow cytometry results showed that apoptosis rate of Siha and He La cells increased after down-regulation of RAGE expression（P<0.05,P<0.01）.Conclusions: Down-regulating the expression of RAGE can inhibit the proliferation and migration of cervical cancer cells and promote the apoptosis of cervical cancer cells.PART Ⅱ Effect of RAGE on sensitivity of cervical cancer cells to cisplatin chemotherapyObjective: To explore the effect of RAGE on the chemotherapy sensitivity of cervical cancer cells to cisplatin and its possible molecular mechanism.Methods: After treatment with different concentrations of cisplatin,the m RNA expression level of RAGE in cervical cancer cells was detected by q PCR.After si-NC and si-RAGE were transfected,the effect of cisplatin on the proliferation of cervical cancer cells was detected by CCK8 assay.After si-NC and si-RAGE were transfected,the effect of cisplatin on apoptosis of cervical cancer cells was detected by flow cytometry.After transfection with si-NC and si-RAGE,the effects of cisplatin on the expression of Caspase3,Bax and Bcl-2 in cervical cancer cells were detected by western blot.Results: The expression level of RAGE m RNA increased with the increase of cisplatin concentration.CCK8 results showed that down-regulation of RAGE can enhance the sensitivity of cervical cancer Si Ha and He La cells to cisplatin（P<0.001,P<0.001）.Flow cytometry showed that down-regulation of RAGE can increase the proportion of cisplatin-induced apoptosis of cervical cancer cells（P<0.05,P<0.01）.Compared with the cisplatin group alone,western blot results showed that the expression levels of Caspase3 and Bax were increased and the level of Bcl-2 was decreased in the cisplatin+si-RAGE group.Conclusions: Inhibition of RAGE can increase the chemotherapy sensitivity of cervical cancer cells to cisplatin.The possible mechanism is that RAGE affects the Bcl-2signaling pathway to enhance the anti-apoptosis ability of cervical cancer cells to cisplatin.