【作者】 赵群； 李勇； 巩涛； 范立桥； 王力利； 焦志凯； 张志栋；

【Author】 Zhao Qun Li Yong Gong Tao etal. The Third Department of Surgery, The Forth Hospital of Hebei Medical University, Shijiazhuang 050011, China

【机构】 河北医科大学第四医院暨河北省肿瘤医院外科；

【摘要】 目的:观察EPA和DHA对人胃癌BGC-823细胞凋亡和G α-PKC βⅡ信号传导的影响。方法:将人胃癌 BGC-823细胞分为对照组,EPA和DHA用药组,EPA组和DHA组按15 μg／ml、30 μg／ml、45μg／ml加药,肿瘤细胞处理48小时后,采用MTT法测定肿瘤细胞抑制率, 细胞抑制率=(1-实验组OD值／对照组OD值)×100％。根据实验结果的观察,选择45μg／ml药物浓度和48h时间点做流式细胞检测肿瘤细胞凋亡率和bcl-2(鼠抗人bcl-2, 1:50)、G α(兔抗人G α q抗体,1:50)、PKC βⅡ基因蛋白的表达(鼠抗人PKC βⅡ抗体,1:50),凋亡率=(凋亡细胞数／所测细胞总数)×100％。凋亡相关基因蛋白表达以荧光指数(FI)表示,FI=(样品蛋白表达的平均荧光强度／正常对照样品平均荧光强度)×100％。应用SPSS13统计软件的单因数方差分析。结果:(1)MTT法测定肿瘤细胞抑制率:根据实验结果的观察,选择45μg／ml药物浓度和48h时间点做进一步实验,人胃癌细胞抑制率在对照组为 0、EPA组为43．06±3．85、DHA组为47．95±3．50,药物组与对照组比较,肿瘤细胞生长均明显受抑制(P<0．001)。(2) 流式细胞测定45μg／ml的EPA和DHA作用48h对肿瘤细胞凋亡率和相关基因蛋白表达的影响如下:①凋亡率对照组为2．53±0．13,EPA组为7．53±0．17,DHA组为 15．24±0．45,EPA和DHA用药组与对照组比较凋亡率明显升高(P<0．001);②bcl-2基因蛋白表达对照组为2．18 ±0．20,EPA组为1．53±0．08,DHA组为1．47±0．13, EPA组和DHA组与对照组比较明显下降(P<0．05);③G α基因蛋白表达对照组为2．95±0．35,EPA组为1．97± 0．22,DHA组为1．85±0．19,EPA组和DHA组与对照组比较明显下降(P<0．001);④PKC βⅡ基因蛋白表达对照组为4．71±0．92,EPA组为2．08±0．52,DHA组为 1．94±0．46,EPA组和DHA组与对照组比较明显下降(P <0．001)。结论:ω-3脂肪酸可诱导人胃癌BGC-823细胞发生凋亡,信号传导途径G α-PKC βⅡ的改变在细胞凋亡中起到重要的作用。更多还原

【Abstract】 Objaotive: To discuss the effect and mechanism of EPA and DHA on apoptosis and G α -PKC β II signal transduction of gastric cells. Methods: Human gastric BGC-823 cells divided into three groups:control group, EPA group and DHA group. Incubation of human gastric BGC-823 cells with different concentration (15mg/ml, 30mg/ml and 45mg/ml) of EPA and DHA for 48h, then cell viability was detected by MTT assay. Apoptotic rate, bcl-2, G α -PKC β II were detected by flow cytometry at 45mg/ml and 48h point Results: The inhibiting rate of the cell were 0 in control group, 43.06 ± 3.85 in EPA group and 47.95 ± 3.50 in DHA group respectively. Compared the above with control group, the cell viability rate was decreased ( P<0.001, respectively ). The rate of apoptotic cells were 2.53 ± 0.13 in control group, 7.53 ± 0.17 in EPA group and 15.24 ± 0.45 in DHA group respectively. Compared the above with control group, the rate of apoptosis was decreased (P<0.001 ). Bcl-2 protein expression (FI) were 2.18 ± 0.20 in control group, 1.53 ± 0.08 in EPA groups and 1.47 ± 0.13 in DHA group respectively. Compared the above with control group (2.18 ± 0.20 ), bcl-2 was decreased ( P<0.001). G a q protein expression ( FI) were 2.95 ± 0.35 in control group, 1.97 ± 0.22 in EPA group and 1.85 ± 0.19 in DHA group respectively.Com-pared the above with control group (2.95 ± 0.35), the G α q protein expression was decreased ( P<0.001 ). PKC β II protein expression were 4.71 ± 0.92 in control groups, 2.08 ± 0.52 in EPA group and 30mg/ml and 1.94 ± 0.46 in DHA GROUP respectively. Compared the above with control group (4.71 ± 0.92 ), PKC β II protein expression was decreased (P<0.001 ).更多还原