β-雌二醇对宫颈癌Hela细胞增殖的影响

doi:10.6040/j.issn.1671-7554.0.2022.0807

摘要/Abstract

摘要： 目的探讨不同浓度β-雌二醇(E₂)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E₂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E₂对Hela细胞增殖的影响;流式细胞仪检测E₂对细胞凋亡和周期的影响;Transwell法检测E₂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E₂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E₂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。

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

Abstract: Objective To investigate the effects of different concentrations of β-estradiol(E₂)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 E₂ on the proliferation were detected with CCK8 assay and cell clone formation assay. The effects of E₂ 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 E₂ 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 E₂ 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.

Key words: Estrogen, Cell cycle, Cervical cancer cells, Cytological behavior, Human papillomavirus

中图分类号:

R737.33

刘萌,侯丛哲,麻焕玉,张震,赵新蕊,张萍,朱琳. β-雌二醇对宫颈癌Hela细胞增殖的影响[J]. 山东大学学报 (医学版), 2023, 61(2): 9-15.

LIU Meng, HOU Congzhe, MA Huanyu, ZHANG Zhen, ZHAO Xinrui, ZHANG Ping, ZHU Lin. Effects of β-estradiol on the proliferation of cervical cancer Hela cells[J]. Journal of Shandong University (Health Sciences), 2023, 61(2): 9-15.

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