二烯丙基二硫对人胃癌细胞RORα蛋白表达的影响

Effect of diallyl disulfide on RORα protein expression in human gastric cancer cells

目的研究二烯丙基二硫(DADS)处理前后人胃癌细胞中维甲酸相关孤核受体α(R ORα)蛋白表达变化情况。方法实验分对照组和DADS处理组,进行体外细胞培养及细胞形态学观察,采用免疫细胞化学、Western blot分析RORα蛋白水平的表达;半定量RT-PCR检测RORα核酸水平的表达。结果DADS处理24 h后,细胞形态学发生明显改变;R ORα蛋白在人胃癌MKN28、MGC803细胞核中呈强阳性表达,与对照组相比表达明显上调;半定量RT-PCR灰度扫描定量分析结果显示,DADS处理24 h后,RORα蛋白表达在MGC803、MKN28细胞中的(0.97±0.01)、(0.91±0.01)较对照组的(0.44±0.02)、(0.72±0.01)显著上调(P<0.01或P<0.05);Western blot分析及灰度扫描显示,DADS处理人胃癌MGC803、MKN28细胞8、12和24 h后,RORα蛋白表达分别为(0.71±0.03)和(0.79±0.01)、(0.87±0.01)和(0.88±0.02)以及(1.31±0.01)和(0.96±0.02)较未处理组的(0.32±0.01)和(0.68±0.02)明显上调,差异有显著性(P<0.01或P<0.05)。结论DADS可诱导人胃癌细胞分化,其诱导分化作用可能与上调R ORα蛋白表达有关。

【Objective】 To investigate the effect of diallyl disulfide（DADS） on retinoic acid-related orphan receptor-alpha（RORα） expression in human gastric cancer cells.【Methods】 To observe morphological changes of the control group and DADS treated group.Immunocytochemistry and western blot were used to observe expression of ROR alpha protein in human gastric cancer cells by DADS.Semi-Quantitative RT-PCR was used to observe expression of nucleic acid level.【Results】 After MGC803 and MKN28 cells treated with DADS for 24 hours,the obvious changes were found under the inverted microscope and light microscope.Immunocytochemical stain indicated that positive expression of RORα increased compared with the control.RT-PCR analysis showed that the RORα expression was up-regulated,the gray scale value ratio of ROR α / β-actin were（0.97±0.01） and（0.91±0.01） in MGC803 and MKN28,compare with（0.44±0.02） and（0.72±0.01） as the control（P〈0.01 or P〈0.05）.Western blot showed that the ROR alpha expression was increased after human gastric cancer cells treated with 30 mg/L DADS for 8 h,12 h and 24 h,the gray scale value ratio of RORα/ β-Actin were（0.71±0.03）、（0.79±0.01）,（0.87±0.01）、（0.88±0.02）,（1.31±0.01）、（0.96±0.02） in MGC803 and MKN28 respectively,compared with（0.32±0.01）,（0.68±0.02） as the control（P〈0.01 or P〈0.05）.【Conclusions】 DADS can induce human gastric cancer MGC803 and MKN28 cells differentiation through up-regulating RORα protein.