摘要
目的金雀异黄酮对多种肿瘤有抑制作用。文中研究金雀异黄酮对宫颈癌Hela细胞增殖的抑制作用和对自噬的诱导,以及自噬在此过程中的作用机制。方法体外培养人宫颈癌Hela细胞,将其分为对照组、金雀异黄酮组和3-甲基腺嘌呤(3-MA)+金雀异黄酮组,对照组只加培养液,金雀异黄酮组用(25、50、100μmol/L)金雀异黄酮处理,3-MA+金雀异黄酮组先用5 mmol/L 3-MA预处理1 h后加入100μmol/L金雀异黄酮,采用MTT法检测各组细胞增殖抑制率的变化;透射电镜观察细胞自噬超微结构的改变;Western blot检测自噬相关蛋白P62及Beclin-1的表达情况;免疫荧光法检测LC3A/B的表达,分析金雀异黄酮对细胞自噬水平的影响。结果与对照组Hela细胞增殖抑制率(0)相比,25、50、100μmol/L金雀异黄酮处理后,Hela细胞增殖的抑制率(20.9%±1.3%,33.5%±1.6%,46.5%±3.2%)逐渐增高(P<0.01)。使用不同浓度(25、50、100μmol/L)金雀异黄酮处理Hela细胞48 h后,随着金雀异黄酮浓度增高Beclin-1表达逐渐增强,P62的表达逐渐降低。激光共聚焦显微镜下观察可见,100μmol/L金雀异黄酮处理组He La细胞细胞质中可见到大量LC3A/B绿色荧光分布,且荧光强度较强。金雀异黄酮(100μmol/L)处理48 h后细胞质中可见大量空泡状结构及包含部分细胞质成分形成双层膜结构的自噬体。金雀异黄酮组Hela细胞的增殖抑制率较3-MA+金雀异黄酮组低[(46.5±3.2)%vs(58.2±2.2)%,P<0.01]。结论金雀异黄酮可抑制Hela细胞增殖,并诱导自噬增强。
Objective For Genistein has been proliferation inhibition to Hela cells by Genistein and the cancer Hela cells were divided into control group, Geniste reported to inhibit many tumors , we investigate the role of autophagy in the machanism of autophagy plays in this process. Methods Human cervical in group and 3-MA＋Genistein group, the control group were cultured in RPMI1640 medium supplemented with 10% fetal bovine serum （FBS）, Genistein group were euhured in various concentrations Genistein（ 25, 50,100 μmol/L）,3-MA＋Genistein group were treated with 5 mmol/L 3-MA for lh before euhured in 100 μmol/L Genistein. The prolifera- tion inhibitory rate of Hela eells was detected by MTr method. The ul- trastrueture changes of Hela cells was observed under transmission e- lectronic microscope （TEM）. The levels of antophagy-assoeiated pro- tein P62 and Beelin-1 were detected by Western blotting analysis. The expressions of autophagy-assoeiated proteins LC3A/B in Hela cellswere determined by fluorescent staining to analyse the autophagy induced by Genistein in Hela cells. Results Compared with control group , the proliferation inhibitory rate of Hela cells was 20.9%± 1.3%, 33.5%± 1.6% and 46.5%± 3.2% when cultured in 25,50,100 μmol/L Genistein（ P〈0.01 ） . After treated with various concentrations Genistein for 48h, we observed a dose-dependent increase in the expression of Beclin-1 and decrease of P62.Confocal laser scanning microscopy confirmed the fluorescent density of LC3A/B expres- sion in Hela cells treated with 100 μmol/L Genistein increased significantly as compared with control group.TEM showed there are many vacuoles and double-membrane autophagosomes which involved cytoplasmic components in Hela cells treated with 100 μmol/L Genistein.The proliferation inhibitory rate of Hela cells of Genistein group is decreased as compared with those in 3-MA＋Genistein group[ （46.5±3.2）% vs （58.2±2.2）%,P〈0.01]. Conclusion Genistein could inhibit Hela cells proliferation and induce autophagy.
出处
《医学研究生学报》
CAS
北大核心
2017年第6期579-583,共5页
Journal of Medical Postgraduates
基金
黑龙江省自然科学基金(D201136)
黑龙江省政府博士后启动基金(LRB06-267)
