抑制Sonic hedgehog信号通路对Hela细胞增殖与凋亡的影响  

Inhibition of Sonic hedgehog signaling pathway on Hela cell proliferation and Apoptosis

作　　者：刘洁 朱清 江梅 丁涛 姜经航 曹玉平 王开秀 蔡丽丽 王婷婷 邵帅 LIU Jie;ZHU Qing;JIANG Mei;DING Tao;JIANG Jinghang;CAO Yuping;WANG Kaixiu;CAI Lili;WANG Tingting;SHAO Shuai(Department of Pathology,Jingmen No.2 People′s Hospital,Jingmen 448000,China)

机构地区：[1]湖北省荆门市第二人民医院病理科,湖北荆门 448000 [2]广州医科大学人体解剖教研室,广州 511436 [3]湖北省荆门市第二人民医院生殖医学中心,湖北荆门 448000

出　　处：《实用医学杂志》2019年第21期3281-3285,共5页The Journal of Practical Medicine

基　　金：湖北省自然科学基金项目(编号：2017CFC808,2018CFC859);荆门市科技计划项目(编号：2018YFZD013)

摘　　要：目的探讨抑制Sonic hedgehog(Shh)信号通路对宫颈癌Hela细胞增殖与凋亡的影响以及可能的机制。方法体外培养Hela细胞并分为对照组、Cyclopamine 10μmol组、Cyclopamine 20μmol组、Cyclopamine 40μmol组;以不同剂量的Shh信号通路抑制剂Cyclopamine抑制Shh信号通路后,通过CCK?8检测各组Hela细胞的增殖情况;通过ELISA法检测细胞Caspase?3和Caspase?9含量表达情况;通过Western Blot法检测细胞增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)、Bcl?2和Bax蛋白的表达。结果(1)随着Cyclopamine药物浓度的增大,Shh信号通路抑制情况更加明显;(2)与对照组相比,抑制Shh信号通路后能够明显抑制Hela细胞的增殖、促进凋亡相关因子Caspase?3和Caspase?9表达;(3)与对照组相比,Cyclopamine抑制Shh信号通路后,与增殖有关蛋白PCNA以及抗凋亡蛋白Bcl?2的表达量显著下调(P<0.01),而促凋亡蛋白Bax的表达量升高(均P<0.01)。结论抑制Shh信号通路能够明显抑制宫颈癌Hela细胞的增殖、促进凋亡,其作用可能与PCNA蛋白表达逐渐下降,抑制Hela细胞DNA合成及分裂增殖有关。Objective To investigate the effect of inhibiting Sonic hedgehog(Shh)signaling pathway on the proliferation and the possible mechanism of apoptosis of cervical cancer Hela cells.Methods Hela cells were cultured in vitro and divided into control group,Cyclopamine low dose group(10μmol),medium dose group(20μmol)and high dose group(40μmol).The proliferation of Hela cells was detected by CCK?8 after inhibiting the Shh signaling pathway with different doses of Cyclopamine,which is the inhibitor of Shh signaling pathway.The expression of caspase?3 and caspase?9 were detected by Elisa.The expression of Proliferating Cell Nuclear Antigen(PCNA),Bcl?2 and Bax were detected by Western Blot.Results(1)With the concentration of Cyclopamine increased,the inhibition of Shh signaling pathway became more obvious.(2)Compared with the control group,inhibition of Shh signaling pathway can significantly inhibit the proliferation of Hela cells and promote the expres?sion of apoptosis?related factors caspase?3 and caspase?9.(3)Compared with the control group,the expressions of proliferation?related protein PCNA and anti?apoptotic protein Bcl?2 were significantly down?regulated(P<0.01),while the expressions of pro?apoptotic protein Bax were increased after using Cyclopamine to inhibit the Shh signal?ing pathway(P<0.01).Conclusions Inhibition of Shh signaling pathway can significantly promote the prolifera?tion and apoptosis of Hela cells in cervical cancer,which may be related to the gradual decrease of PCNA protein expression and inhibition of DNA synthesis,division and proliferation of Hela cells.

关 键 词：宫颈癌 Sonic HEDGEHOG信号通路 CYCLOPAMINE HELA细胞 凋亡 

分 类 号：R73[医药卫生—肿瘤]