Research Relationship Between GPR4 And The Proliferation Of Cervical Cancer Cells

Objective:Cervical cancer is one of the malignant tumors that seriously threaten the health of women in our country.The aim of this study is to explore the relationship between GPR4 and the proliferation of cancer cells in cervical cancer,and hope to provide new ideas and targets for the treatment of cervical cancer.It provides experimental basis for the development and search of new drugs.Methods:The plasmid of GPR4 shRNA interference carrier was constructed and the pMSCV vector empty plasmid was constructed with the same principle.Cervical cancer He La cells were divided into three groups: GPR4 shRNA interfering vector plasmid group,pMSCV vector empty plasmid group and simple cervical cancer HeLa cell group.(1)the transfection efficiency of GPR4 interfering gene was detected by fluorescence microscope.(2)the proliferation of cervical cancer Hela cells was analyzed with the method of MTT(four methyl azazolium salt).(3)the cell cycle of each group was detected by flow cytometry,and the apoptosis was analyzed.(4)Western blot method was used to detect the changes of GPR4 protein and VEGF expression in each group.Results:1.The results of MTT l showed that after transfection,GPR4 shRNA interference plasmid group absorbance(OD)value were decreased,cell proliferation is slow,compared with the pMSCV vector empty plasmid group had significant difference(P<0.05),and pMSCV vector empty plasmid group and blank control group compared with no significant difference(P<0.05),suggesting that knockdown of GPR4 gene,human cervical carcinoma cell line Hela cells proliferation rate was inhibited,and the growth curve comparison,GPR4 shRNA interference plasmid group compared with the control group,the growth trend was slow.2.flow cytometry analysis showed that: three groups of cells in each period of detection results found that GPR4 shRNA interference vector transfection group G1 phase cells increased significantly(P<0.05),S phase cells decreased(P<0.05),while empty plasmid control group and blank control group no significant difference(P>0.05).3.flow cytometry,apoptosis of empty plasmid control group and blank control group respectively(3.13 + 0.18)% and(2.91 + 0.23)%,while the group Hela cell apoptosis GPR4 shRNA interference vector transfection rate was(10.74 + 0.38)%,and the empty plasmid control group,blank the control group apoptosis rate comparison,the difference was significant(P<0.05).4.Western blot method was used to determine the expression of GPR4 protein and VEGF in GPR4 shRNA interference vector transfection group.The difference between GPR4 protein and VEGF expression between empty plasmid control group and blank control group was significant(P<0.05).Conclusion:1.GPR4 shRNA interference plasmids inhibit the growth of HeLa cells in cervical cancer.2.GPR4 shRNA interfering plasmids can accelerate the apoptosis of He La cells in cervical cancer.3.GPR4 shRNA interference plasmid can significantly reduce the expression of GPR4 and VEGF protein in cervical cancer HeLa cells,and indicate that the decrease of GPR4 expression affects the decrease of VEGF protein.