二烯丙基二硫对人胃癌细胞维甲酸相关孤核受体α表达的影响

Effect of Diallyl Disulfide on the Expression of Retinoic Acid-related Orphan Receptor-alpha in Human Gastric Cancer Cells

目的探讨二烯丙基二硫(DADS)处理前后人胃癌细胞中维甲酸相关孤核受体α(RORα)表达变化情况。方法 将胃癌SGC7901、MGC803细胞分为对照组和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,胃癌SGC7901、MGC803细胞形态学发生明显改变;RORα在人胃癌SGC7901、MGC803细胞核中呈强阳性表达,与对照组比较表达显著上调;半定量RT-PCR灰度扫描定量分析结果显示,DADS处理24 h后,SGC7901、MGC803细胞中RORα的表达量[(1.09±0.01)与(0.97±0.01)]较对照组[(0.45±0.03)与(0.44±0.02)]显著上调,差异均有统计学意义(P<0.01);Western blot分析及灰度扫描显示,DADS处理人胃癌MGC803、SGC7901细胞8、12、24 h后,RORα蛋白表达量较未处理组显著上调,差异均有统计学意义(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RORα表达有关。

Objective To investigate the effect of diallyl disulfide（DADS） on the expression of retinoic acid-related orphan receptor-alpha（RORα） in human gastric cancer cells. Methods Gastric cells SGC7901 and MGC803 were divided into DADS group and control group,and then the morphological changes were observed after in vitro culture.The expression of RORα protein in human gastric cancer cells was detected by immunocytochemistry and Western blotting.The expression of nucleic acid was detected by semi-quantitative RT-PCR. Results After SGC7901 and MGC803 cells were treated with DADS for 24 hours,obvious morphological changes were found under the inverted microscope and light microscope.Immunocytochemical stain indicated that the positive expression of RORα significantly increased in DADS group than in control group.RT-PCR analysis showed that the RORα expression were up-regulated and the gray scale value ratio of RORα/β-actin were（1.09±0.01） and（0.97±0.01） in SGC7901 and MGC803 in DADS group,compare with（0.45±0.03） and（0.44±0.02） in control group（P〈0.01）.Western blotting showed that RORα expression was significantly increased after human gastric cancer cells treated with 30 mg/L DADS for 8 h,12 h and24 h,compare with those in control group（P〈0.05）. Conclusion DADS can induce the differentiation of human gastric cancer SGC7901 and MGC803 cells via up-regulating RORα protein.