摘要
在肝癌细胞SMMC7721中研究TPA诱导的凋亡及它与细胞核受体TR3和RXRα的关系.实验中使用SMMC7721肝癌细胞.用MTT法检测生长抑制.用DAPI荧光染色法观察凋亡形态,随机记数1 000个细胞以计算凋亡指数.用CAT分析检测RXRα转录活性的抑制.用Western Blot检测RXRα蛋白的表达水平.用RT-PCR检测TR3 mRNA的表达水平.用MTT检测到TPA对SMMC7721肝癌细胞的生长抑制,并且有剂量依赖性.TPA诱导SMMC7721肝癌细胞的凋亡,RXRα蛋白随时间依赖性的降解,并且用CAT分析检测到TPA能抑制RXRα的转录活性.另外TPA能诱导TR3 mRNA的表达上调.TPA抑制SMMC7721肝癌细胞的生长,并诱导凋亡.TPA能上调TR3的mRNA,使RXRα随时间依赖性的降解,并能抑制RXRα的转录活性.因此TPA诱导SMMC7721肝癌细胞的凋亡可能与细胞核受体TR3和RXRα有某些联系.
It is studied that induction of apoptosis by TPA and its relationship with TR3 and RXRα in hepatocellular cell SMMC7721. Hepatocellular cell line, SMMC7721 was used in the experiments. Growth suppression was detected with MTT colorimetric assay. Apoptotic morphology was observed by DAPI fluorescence staining, and apoptotic index was counted among 1 000 cells randomly. Inhibition of RXRα transcriptional activity by TPA was detected by CAT assay. The expreasion level of RXRα protein was detected by Western Blot. The expression level of TR3 mRNA was detected by RT - PCR. Growth suppression was observed with MTT colorimetric assay. The inhibitory effect was dose-dependent. Treatment of SMMC7721 cells with TPA resulted in apoptosis, accompanied by the reprexsion of RXRα protein in a time-dependent manner. At the same time, TPA also up- regulated expression level of TR3 mRNA in SMMC7721 cells that expressed TR3 mRNA. Inhibition of RXRα transcriptional activity by TPA was observed by CAT assay. TPA inhibits proliferation, induces apoptosis of SMMC7721 cells. Expression level of TR3 mRNA in SMMC7721 was induced by TPA. TPA repressed expression level of RXRα protein and inhibited RXRα transcriptional activity. And apoptosis has some relation with TR3 and RXRα.
出处
《云南大学学报（自然科学版）》
CAS
CSCD
北大核心
2006年第3期271-276,共6页
Journal of Yunnan University(Natural Sciences Edition)
基金
国家自然科学基金资助项目(30481939)
