摘要
目的探讨麦冬皂苷(OP)-B增强前列腺癌(PC)-3细胞放疗敏感性及其可能机制。方法体外培养PC-3细胞,PC-3细胞分为5组:空白对照组(未处理)、阳性对照组(Notch1信号通路特异性阻断剂DAPT处理)、低剂量组(加入2.5μmol/L OP-B)、中剂量组(加入5μmol/L OP-B)、高剂量组(加入10μmol/L OP-B),每组细胞平均分成5份,每份只经0、2、4、6、8 Gy其中一种辐射处理48 h后进行后续实验。四甲基偶唑氮盐(MTT)实验检测细胞增殖;流式细胞仪检测细胞凋亡;酶联免疫吸附试验检测乳酸脱氢酶(LDH)漏出率。Western印迹法检测Notch1、Hes1、B细胞淋巴瘤(Bcl)-2、Bcl-2相关蛋白(Bax)、裂解的半胱氨酸天冬氨酸蛋白酶(cleaved Caspase)-3、裂解的多聚ADP核糖聚合酶(cleaved PARP)蛋白表达水平。结果与空白对照组相比,阳性对照组及OP-B不同剂量组PC-3细胞增殖率、LDH漏出率、Notch1、Hes1及Bcl-2蛋白表达水平均显著降低(P<0.05),且高剂量组显著低于低剂量组、中剂量组(P<0.05);与空白对照组相比,阳性对照组及OP-B不同剂量组PC-3细胞凋亡率、Bax、cleaved caspase-3、cleaved PARP蛋白表达水平均显著升高,呈剂量依赖关系(P<0.05),且高剂量组显著高于低剂量组、中剂量组(P<0.05),高剂量组与阳性对照组比较差异无统计学意义(P>0.05)。结论OP-B可促进PC细胞凋亡并增加其放疗敏感性,其作用机制可能与抑制Notch1信号通路有关。
Objective To investigate the effect of Ophiopogonin-B(OP-B)on radiosensitivity of prostate cancer PC-3 cells and its possible mechanism,so as to provide theoretical basis for improving the therapeutic effect of prostate cancer.Methods PC-3 cells were cultured in vitro and divided into five groups:blank control group(untreated);positive control group(treated with DAPT,a specific blocker of Notch1 signaling pathway);low-dose group(added 2.5μmol/L OP-B);medium-dose group(added 5.0μmol/L OP-B);high-dose group(added 10.0μmol/L OP-B),cells in each group were divided into five parts,each of which was treated with 0,2,4,6 and 8 Gy for 48 hours.Tetramethyl azo salt(MTT)assay was used to detect cell proliferation;flow cytometry was used to detect cell apoptosis;the leakage rate of lactate dehydrogenase(LDH)was detected by ELISA.Western blot was used to detect the expression levels of Notch1,Hes1,B-cell lymphoma-2(Bcl-2),Bcl-2 related protein(Bax),cleaved caspase-3,and cleaved ADP ribose polymerase(cleaved PARP).Results Compared with the blank control group,the PC-3 cell proliferation rate,LDH leakage rate,Notch1,Hes1 and Bcl-2 protein expression levels in the positive control group and OP-B different dose groups were significantly lower(P<0.05),those of the high-dose group were significantly lower than those of the low-dose group and the middle-dose group(P<0.05).Compared with the blank control group,the apoptotic rate of PC-3 cells,the expressions of Bax,cleaved caspase-3,and cleaved PARP proteins in the positive control group and OP-B different doses groups were significantly increased in a dose-dependent manner(P<0.05),those of the high-dose group were significantly higher than those of the low-dose group and the middle-dose group(P<0.05),there were no significant differences between high-dose group and positive control group in above indexes(P>0.05).Conclusions OP-B could promote apoptosis of prostate cancer cells and increase their radiosensitivity,its mechanism might be related to the inhibition of Notch1 signaling pathway.
作者
李挺云
梁培育
王声兴
周治彦
陈金火
LI Ting-Yun;LIANG Pei-Yu;WANG Sheng-Xing(Department of Urology,the First Affiliated Hospital of Hainan Medical College,Haikou 570102,Hainan,China)
出处
《中国老年学杂志》
CAS
北大核心
2020年第21期4626-4631,共6页
Chinese Journal of Gerontology
基金
海南省社会发展科技专项项目(No.2015SF31)。
