白藜芦醇抑制胃癌细胞株MKN45增殖的机制研究*

Resveratrol inhibits proliferation of gastric cancer MKN45 cells by regulating the MEK/ERK*

摘要:

目的 通过研究白藜芦醇抑制胃癌MKN45细胞增殖及对丝裂原活化蛋白激酶(MEK)/细胞外信号调节激酶（ERK）信号通路的影响，探讨白藜芦醇抗胃癌的作用机制。方法 体外常规培养胃癌MKN45细胞，将细胞分为对照组、MEK/ERK信号通路抑制剂PD98059组（PD98059组）、白藜芦醇低剂量组（100?μmol/L）、 白藜芦醇高剂量组（400?μmol/L）。各组细胞处理1、2和4?h后，MTT法检测细胞增殖抑制率，Western blotting法检测Ras、Raf、MEK、ERK1/2蛋白的表达，细胞免疫化学法检测Ras、Raf、MEK、ERK1/2蛋白荧光强度的变化。结果 与对照组比较，PD98059组、白藜芦醇低剂量组和白藜芦醇高剂量组细胞在处理1、2和4?h后，细胞增殖受到抑制（P?<0.05），且呈时间剂量依赖性；PD98059组、白藜芦醇低剂量组和白藜芦醇高剂量组细胞Ras、Raf、MEK和ERK1/2表达下降，差异有统计学意义（P?<0.05），且白藜芦醇的作用呈剂量依赖性（P?< 0.05）；MEK和ERK1/2蛋白主要定位于细胞质中，且PD98059组、白藜芦醇低剂量组和白藜芦醇高剂量组细胞ERK1/2的荧光强度均减弱。结论 白藜芦醇呈剂量依赖性抑制胃癌细胞株MKN45的增殖，可能与抑制MEK/ERK信号通路，减弱Ras、Raf、MEK和ERK1/2的表达有关。

Abstract:

Objective To investigate the inhibition effect and mechanism of resveratrol on proliferation of gastric cancer MKN45 cell and the involvement of MEK/ERK signaling pathway. Methods Gastric cancer MKN45 cells were cultured according to the conventional signaling pathway method. The experiment was divided into control group, MEK/ERK signaling pathway inhibitor PD98059 group (20?mg/L, 100?μl), resveratrol low-dose group (100?μmol/L), resveratrol high-dose group (400?μmol/L). Gastric cancer MKN45 cell was treated with 1?h, 2?h and 4?h for subsequent follow-up. MTT assay was used to detect the cell viability of the inhibition rate of cell proliferation. Western blotting was used to analyze the expression of proteins such as Ras, Raf, MEK, ERK1/2. Immunocytochemistry was used to analyze the fluorescence intensity of Ras, Raf, MEK and ERK1/2. Results Compared with control group, the proliferation of cells in PD98059 group, low dose resveratrol group and high dose resveratrol group was significantly inhibited after treatment for 1?h, 2?h and 4?h with a time-dependent and dose-dependent way (P?