摘要
目的研究汉防己甲素对宫颈癌细胞增殖、凋亡和放射敏感性的影响,并探讨其作用机制。方法以不同浓度(0.01μg/ml、0.10μg/ml、1.00μg/ml、5.00μg/ml、10.00μg/ml)汉防己甲素处理Hela细胞,MTT法检测细胞增殖;5.00μg/ml汉防己甲素处理Hela细胞48 h后,流式细胞仪检测细胞凋亡率,qRT-PCR和western blot检测Chk1的表达,克隆形成实验检测各组细胞放射敏感性。转染Chk1 siRNA至Hela细胞,检测细胞增殖、凋亡和放射敏感性。转染pcDNA-Chk1至Hela细胞,并用5.00μg/ml汉防己甲素处理48 h,检测细胞增殖、凋亡和放射敏感性。结果汉防己甲素能明显抑制Hela细胞的增殖,且呈浓度依赖性;与对照相比,5.00μg/ml汉防己甲素可明显提高细胞凋亡率,增强Hela细胞放射敏感性,抑制Chk1的表达(P<0.05)。沉默Chk1基因可显著抑制Hela细胞的增殖,促进细胞凋亡,增强放射敏感性(P<0.05)。过表达Chk1逆转了汉防己甲素对Hela细胞增殖、凋亡和放射敏感性的影响。结论汉防己甲素可抑制Hela细胞的增殖,促进细胞凋亡,增强细胞放射敏感性,其机制可能与调控Chk1的表达有关,将可为宫颈癌的治疗提供新依据。
Objective To investigate the effects of tetrandrine on proliferation,apoptosis and radiosensitivity of cervical cancer cells by regulating the expression of Chk1 genein vitro,and to explore its action mechanism.Methods Hela cells were treated with different concentration of tetrandrine(0.01μg/ml,0.10μg/ml,1.00μg/ml,5.00μg/ml,10.00μg/ml),then the proliferation of Hela cells was detected by MTT assay.After treating Hela cells with 5.00μg/ml tetrandrine for 48h,the apoptosis rate was detected by flow cytometry,the expression levels of Chk1 were detected by qRT-PCR and Western Blot,and the radiosensitivity was detected by clonal formation assay.Moreover Chk1 siRNA was transfected into Hela cells,then the cell proliferation,apoptosis and radiosensitivity were detected.And pcDNA-Chk1 was transfected into Hela cells and treated with tetrandrine at 5.00μg/ml for 48h,and the cell proliferation,apoptosis and radiosensitivity were detected.Results The tetrandrine inhibited obviously the proliferation of Hela cells in a dose-dependent manner.Moreover 5.00μg/ml tetrandrine significantly increased the apoptosis rate and the radiosensitivity of Hela cells,and inhibited the expression of Chk1(P<0.05).Silencing Chk1 gene could significantly inhibit the proliferation of Hela cells,promote apoptosis and radiosensitivity(P<0.05).Moreover the overexpression of Chk1 reversed the effects of tetrandrine on proliferation,apoptosis and radiosensitivity of Hela cells.Conclusion The tetrandrine can inhibit the proliferation of Hela cells,promote cell apoptosis,and increase the radiosensitivity,and itc action mechanism may be related to the regulation of Chk1 expression,which may provide a new basis for the treatment of cervical cancer.
作者
龚妙添
杨桂生
黄仕尧
GONG Miaotian;YANG Guisheng;HUANG Shiyao(Department of Clinical Laboratory,Sanshui Distict Hospital for Maternal and Child Health,Guangdong,Foshan 528100,China;不详)
出处
《河北医药》
CAS
2021年第20期3078-3081,3086,共5页
Hebei Medical Journal
基金
广东省科技计划项目(编号:20160212)。
