| Objective:Persistent infection of high-risk human papillomavirus?HPV?is the causing the majority of cervical cancers,but not all patients infected with high-risk HPV eventually onset of cervical cancer.This suggests that there probably exist synergies promoting the occurrence of cervical cancer.These synergies may include genetic predisposition,changes in topical vaginal microenvironment,and the effect of estrogen.In this study,the effects of 17?-estradiol?17?-E2?on the proliferation and migration of cervical squamous cell carcinoma of HPV16-negative C33A and HPV16-positive SiHa cells were compared to explore the differences in the effects of estrogen on HPV positive and HPV negative cervical squamous cell carcinoma.Methods:HPV16-negative C33A and HPV16-positive SiHa cells of cervical squamous cell carcinoma were cultured in vitro,and logarithmic cells were taken for the experiments.Blank control groups?excluding 17?-E2?and17?-E2 groups were set up in the experiment.The peak concentration of drug action was screened by CCK8 method,cell cycle distribution of each group was detected by flow cytometry,Cyclin D1,PCNA protein expression level of each group of cells was detected by Western blot method,and cell migration ability was detected by cell scratch test.Results:1.17?-E2 promotes viability of HPV16-negative C33A and HPV16-positive SiHa cellsCCK-8 kit was used to detect the effect of 17?-E2 on the viability of C33A and SiHa cells.Combined with existing reports,two cells were treated for 72 h,and different doses of 17?-E2(0,10-10,10-9,10-8,10-7mol/L)can enhance cell viability.The results showed that the viability of C33A and SiHa cells increased in a dose-dependent manner with the increase of the 17?-E2concentration,and the viability of SiHa cells was significantly higher than that of C33A cells.For subsequent experiments,10-7 mol/L will be selected as the experimental concentration.2.17?-E2 promotes cell proliferation by regulating cell cycle progression,SiHa cells have a strong proliferation activityAfter treatment with 17?-E2(10-7mol/L)for 72 h,the number of C33A and SiHa cells in S and G2/M phases increased,and the number of G0/G1cells decreased in comparison with the control group.Among them,the number of Si-phase cells in SiHa cells increased more significantly than that in C33A cells.Western blot results showed that,compared with the control group,the treatment of 17?-E2 with two cells significantly increased the expression of proliferation-related proteins Cyclin D1 and PCNA for 72 h,and the increase of Cyclin D1 and PCNA was more significant in SiHa cells than in C33A cells.In summary,the results suggest that 17?-E2 promotes cell proliferation by regulating cell cycle progression,and SiHa cells have a strong proliferation activity.3.17?-E2 can enhance the migration ability of C33A and SiHa cells,SiHa cells have better migration ability than C33AAfter treatment with 17?-E2 for 72 h,the scratches of the two cells in the17?-E2 group were covered by the cells that migrated to the center,and the remaining scratch area ratio was significantly reduced compared with the control group.The scratch test results showed that 17?-E2 can significantly enhance the migration ability of C33A and SiHa cells,while the migration ability of SiHa cells is stronger than that of C33A cells.Conclusions:1.Estrogen can promote the proliferation and migration of HPV16-positive SiHa cells and HPV16-negative C33A cells in cervical squamous cell carcinoma.2.The effect of estrogen on cervical squamous cell carcinoma HPV16positive SiHa cells is stronger than that of estrogen on cervical squamous cell carcinoma HPV16 negative C33A cells.It is speculated that estrogen may promote cell proliferation by cooperating with HPV oncogenes.3.The mechanism of estrogen in the development of cervical cancer needs to be further explored. |
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