冬凌草甲素通过G2/M期阻滞和诱导细胞凋亡增强人胃癌细胞放疗敏感性  

Oridonin facilitates the radiosensitivity of gastric cancer via inducting G2/M phase arrest and apoptosis

作　　者：高灵犀 张妍 汪文 张腾 GAO Lingxi;ZHANG Yan;WANG Wen;ZHANG Teng(Department of Gastroenterology,The First Affiliated Hospital of Wannan Medical College,Wuhu 241001,China)

机构地区：[1]皖南医学院第一附属医院弋矶山医院消化内科,安徽芜湖241001

出　　处：《皖南医学院学报》2022年第4期315-320,共6页Journal of Wannan Medical College

基　　金：安徽高校自然科学研究重点项目(KJ2020A0594)。

摘　　要：目的:研究冬凌草甲素增强放疗敏感性的潜在机制。方法:CCK-8实验检测SGC7901人胃癌细胞经不同放疗剂量和不同冬凌草甲素浓度各处理72 h后细胞增殖情况。EdU法检测冬凌草甲素和(或)放疗对SGC7901人胃癌细胞增殖的影响。平板克隆实验检测冬凌草甲素和(或)放疗对SGC7901细胞克隆生长能力的影响。流式细胞技术检测SGC7901人胃癌细胞凋亡和细胞周期。qPCR检测Apaf-1、Bcl-2、Bax、Caspase-3、Cytochrome C的mRNA表达。Western blot检测Apaf-1、Bcl-2、Bax、Cleaved-caspase3、Cytochrome C蛋白表达。结果:CCK-8实验结果显示细胞活力随放疗剂量和冬凌草甲素浓度的升高而下降。平板克隆形成实验及EdU细胞增殖实验结果显示联合组细胞增殖率低于放疗组(P<0.05)。细胞周期实验结果显示冬凌草甲素组G2/M期细胞比例高于空白对照组;联合组G2/M期细胞比例高于放疗组(P<0.05)。细胞凋亡实验、qPCR及Western blot结果显示冬凌草甲素组和放疗组细胞凋亡率高于空白对照组(P<0.05),与空白对照组相比,两组凋亡相关蛋白Apaf-1、Bax、Cleaved-caspase3、Cytochrome C表达上升(P<0.05),抗凋亡蛋白Bcl-2表达下降(P<0.05);联合组细胞凋亡率高于放疗组(P<0.05),凋亡相关蛋白Apaf-1、Bax、Cleaved-caspase3、Cytochrome C表达上升较放疗组更明显(P<0.05),抗凋亡蛋白Bcl-2表达下降较放疗组更显著(P<0.05)。结论:冬凌草甲素通过G2/M期阻滞和诱导细胞凋亡增强人胃癌细胞放疗敏感性。Objective:To investigate the potential mechanism of oridonin in enhancing radiotherapy sensitivity.Methods:CCK-8 was used to detect the cell proliferation of gastric cancer cell SGC7901 following different dose of radiotherapy and oridonin concentration for 72 h.Proliferation of SGC7901 cells treated with oridonin and/or radiotherapy was detected by EdU assay.Plate cloning assay was performed to measure the clonal growth ability of SGC7901 cells after treatment with oridonin and/or radiotherapy.Apoptosis and cell cycle of gastric cancer cells were detected by flow cytometry.mRNA expression of Apaf-1,Bcl-2,Bax, caspase-3 and cytochrome C was determined using qPCR.Western blot was used to detect the protein expression of Apaf-1,Bcl-2,Bax, cleaved caspase-3,cytochrome C.Results:CCK-8 assay showed that the cell viability was decreased with added radiotherapy dose and oridonin concentration.The results of plate clone formation test and EdU cell proliferation assay showed that the cell proliferation rate was significantly lower in the combined treatment group than in single radiotherapy group(P<0.05).Cell cycle experiment demonstrated that the proportion of G2/M cells in oridonin group was higher than that in blank control group, and the proportion of G2/M phase cells was higher in the combined treatment group than in the radiotherapy group(both P<0.05).Apoptosis experiment, qPCR and Western blot showed that the apoptosis rate in oridonin group and radiotherapy group was higher than that of blank control group(P<0.05).The expression of apoptosis related proteins Apaf-1,Bax, cleavedcaspase-3 and cytochrome C was increased(P<0.05),yet the expression of anti-apoptotic protein Bcl-2 was decreased(P<0.05).The apoptosis rate in the combined treatment group was higher than that in the radiotherapy group(P<0.05).The expression of apoptosis related proteins Apaf-1,Bax, cleaved caspase-3 and cytochrome C was more dominantly increased, yet anti-apoptotic protein Bcl-2 was more significantly decreased compared to single radiothera

关 键 词：胃癌 冬凌草甲素 放疗敏感性 细胞周期 凋亡 

分 类 号：R285.5[医药卫生—中药学] R735.2[医药卫生—中医学]