棕矢车菊素诱导人宫颈癌Hela细胞G_(2)/M期停滞的机制研究  

The mechanism of G_(2)/M arrest induced by Jaceosidin in human cervical cancer Hela cells

作　　者：王海艳[1] 秦海霞[2] 李淑娟[1] WANG Haiyan;QIN Haixia;LI Shujuan(Gynecology and Obstetrical Department,Anyang District Hospital of Puyang City,Henan Anyang 455000,China;Department of Gynaecology,the First Affiliated Hospital of Xinxiang Medical College,Henan Xinxiang 453000,China)

机构地区：[1]濮阳市安阳地区医院妇产科,河南安阳455000 [2]新乡医学院第一附属医院妇科,河南新乡453000

出　　处：《现代肿瘤医学》2022年第18期3264-3269,共6页Journal of Modern Oncology

基　　金：河南省高等学校重点科研项目计划(编号:19A320030)。

摘　　要：目的:研究棕矢车菊素对宫颈癌(cervical carcinoma,CC)细胞Hela的增殖抑制和周期阻滞能力,探索棕矢车菊素是否具有宫颈癌治疗药物的潜力。方法:MTT比色法检测细胞活力;羧基荧光素二醋酸盐琥珀酰亚胺酯(carboxyfluorescein succinimidyl amino ester,CFSE)法检测细胞增殖水平变化;碘化丙啶(propidium,PI)单染检测细胞周期;免疫印迹法(western blot,WB)检测细胞蛋白表达水平。采用裸鼠成瘤检测棕矢车菊素体外抑制肿瘤效果。结果:棕矢车菊素可以显著抑制Hela细胞活力水平,且具有时间和浓度依赖性;棕矢车菊素抑制Hela细胞增殖,且100μmol/L棕矢车菊素可显著诱导细胞发生G_(2)/M周期阻滞。分子机制上,研究证明了棕矢车菊素可以通过促进细胞周期检验点激酶(checkpoint kinase 2,CHK2)激活,抑制细胞分裂周期蛋白25同源蛋白C(cell division cyclin 25 homolog C,CDC25C)活力,从而提高无活性的pCDC2水平。棕矢车菊素同时降低了Cyclin B1表达。因此,棕矢车菊素通过抑制G_(2)/M期调控蛋白Cyclin B1/CDC2复合物活性来诱导细胞发生G_(2)/M期阻滞。同时,体外裸鼠成瘤,棕矢车菊素灌胃4周后,小鼠肿瘤体积显著减小。结论:在Hela细胞上,棕矢车菊素具有抑制细胞增殖和诱导G_(2)/M期阻滞的作用,证明了棕矢车菊素具有抗宫颈癌细胞的治疗潜力。Objective:To determine the proliferation inhibition and cell cycle arrest ability of Jaceosidin in cervical carcinoma(CC)Hela cells and investigate the potential of Jaceosidin to be the novel agent for CC treatment.Methods:MTT was used to determine the cell viability.Cell proliferation was detected by carboxyfluorescein succinimidyl amino ester(CFSE)and flow cytometry.Propidium(PI)-single staining was performed to detect cell cycle arrest.Western blot(WB)was used to determine the expression of proteins.The tumor inhibiting effect of palmiflorin in vitro was detected in nude mice.Results:Jaceosidin inhibited the cell viability of Hela cells in a time-and concentration-dependent manner.Jaceosidin inhibited cell proliferation and 100μmol/L Jaceosidin induced G_(2)/M phase cell cycle arrest significantly.In mechanism,100μmol/L Jaceosidin promoted activation of checkpoint kinase 2(CHK2)and inhibited the activation of cell division cyclin 25 homolog C(CDC25C),subsequently enhanced the expression of pCDC2.Besides,Jaceosidin decreased the expression of Cyclin B1.Thus,Jaceosidin induced G_(2)/M phase cell cycle arrest by inhibition of Cyclin B1/CDC2 activation.At the same time,tumor was formed in nude mice in vitro.After 4 weeks of intragastric administration,the tumor volume of mice was significantly reduced.Conclusion:In Hela cells,Jaceosidin inhibited cell proliferation and induced G_(2)/M cell cycle arrest,suggesting the potential of Jaceosidin on CC treatment.

关 键 词：HELA细胞 细胞增殖 G_(2)/M 细胞周期 棕矢车菊素 

分 类 号：R737.33[医药卫生—肿瘤]