β-雌二醇对宫颈癌Hela细胞增殖的影响  

Effects of β-estradiol on the proliferation of cervical cancer Hela cells

作　　者：刘萌 侯丛哲 麻焕玉 张震 赵新蕊 张萍[1] 朱琳[1] LIU Meng;HOU Congzhe;MA Huanyu;ZHANG Zhen;ZHAO Xinrui;ZHANG Ping;ZHU Lin(Department of Gynecology,The Second Hospital of Shandong University,Jinan 250033,Shandong,China)

机构地区：[1]山东大学第二医院妇科,山东济南250033

出　　处：《山东大学学报（医学版）》2023年第2期9-15,共7页Journal of Shandong University:Health Sciences

基　　金：山东省自然科学基金(ZR2020MH069);济南市科学技术计划项目(201907008);济南市科学技术计划项目(201907009)。

摘　　要：目的探讨不同浓度β-雌二醇(E_(2))对宫颈癌Hela细胞增殖的影响及机制。方法体外培养Hela细胞,根据E_(2)的浓度将其分为对照组、低浓度组、高浓度组。利用CCK8法和细胞克隆形成试验检测E_(2)对Hela细胞增殖的影响;流式细胞仪检测E_(2)对细胞凋亡和周期的影响;Transwell法检测E_(2)对细胞迁移能力的影响;在对照组和高浓度组中,利用实时定量PCR(qRT-PCR)检测HPV18 E6、E7、p21、p53、CDK2、Rb基因的表达,Western blotting检测p21、p53、CDK2、Rb、Rb磷酸化(pRb)蛋白的表达。结果相比于对照组及低浓度组,高浓度组中细胞增长率降低(P<0.001),细胞克隆形成数量被抑制(P=0.013),细胞周期G0/G1期延长(P<0.001)。高浓度E_(2)可降低HPV18 E6/E7 mRNA表达水平(P=0.049,P=0.043),并下调CDK2、Rb mRNA和蛋白表达水平(P_(CDK2 mRNA)=0.035,P_(Rb mRNA)=0.003,P_(CDK2蛋白)=0.009,P_(Rb蛋白)=0.008),上调p21、p53 mRNA和蛋白表达水平(P_(p21 mRNA)=0.042,P_(p53 mRNA)=0.035,P_(p21蛋白)=0.039,P_(p53蛋白)=0.043),同时下调pRb蛋白表达水平(P<0.001)。结论高浓度E_(2)能够抑制Hela细胞增殖,其机制可能与抑制HPV18 E6/E7癌基因、促进p53和p21蛋白的表达、调节细胞周期蛋白(CDK2、Rb)的水平进而影响正常细胞周期进程有关。Objective To investigate the effects of different concentrations ofβ-estradiol(E_(2))on the proliferation of Hela cells in cervical cancer and the mechanism.Methods Hela cells cultured in vitro were divided into control,low concentration and high concentration groups.The effects of E_(2) on the proliferation were detected with CCK8 assay and cell clone formation assay.The effects of E_(2) on the apoptosis,cell cycle and migration were detected with flow cytometry and Transwell assay,respectively.In the control and high concentration groups,the mRNA expressions of HPV18 E6,E7,p21,p53,CDK2 and Rb were detected with qRT-PCR;the protein expressions of p21,p53,CDK2,Rb and Rb phosphorylation(pRb)were detected with Western blotting.Results Compared with the control and low concentration groups,the high concentration group had reduced cell growth rate(P<0.001),inhibited cell cloning(P=0.013)and prolonged G0/G1 phase(P<0.001).High concentration of E_(2) reduced the mRNA expressions of HPV18 E6/E7(P=0.049,P=0.043),downregulated the mRNA and protein expressions of CDK2 and Rb(P_(CDK2 mRNA)=0.035,P_(Rb mRNA)=0.003,P_(CDK2 protein)=0.009,P_(Rb protein)=0.008),upregulated the mRNA and protein expressions of p21 and p53(P_(p21 mRNA)=0.042,P_(p53 mRNA)=0.035,P_(p21 protein)=0.039,P_(p53 protein)=0.043),and downregulated pRb(P<0.001).Conclusion High concentration of E_(2) inhibits the proliferation of Hela cells.The mechanism might be related to the inhibition of HPV18 E6/E7 oncogene,promotion of the protein expressions of p53 and p21,and regulation of cell cycle protein(CDK2,Rb)levels,thus affecting the normal cell cycle.

关 键 词：Β-雌二醇 细胞周期 宫颈癌细胞 细胞学行为 人乳头瘤病毒 

分 类 号：R737.33[医药卫生—肿瘤]