全反式维甲酸对肝癌细胞连接蛋白基因表达及细胞间隙连接通讯功能的影响  

Effects of all-trans retinoic acid on expression of connexin genes and gap junction communication in hepatocellular carcinoma cell lines

作　　者：刘颖斌[1] 许斌[2] 王建伟[1] 方河清[1] 李江涛[1] 李海军[1] 唐喆[1] 钱浩然[1] 冯雪冬[1] 彭淑牖[1] 

机构地区：[1]浙江大学医学院附属第二医院,杭州310009 [2]浙江大学医学院附属邵逸夫医院

出　　处：《中华医学杂志》2005年第20期1414-1418,共5页National Medical Journal of China

摘　　要：目的观察肝癌细胞株SMMC-7721和BEL-7404经全反式维甲酸(ATRA)诱导前后CX26、CX32、CX43基因在转录水平的改变以及对细胞间隙连接功能的影响。方法分别用常规培养基和含ATRA浓度为10-5mol/L的培养基培养SMMC-7721和BEL-7404两种肝癌细胞株细胞,于药物诱导细胞24、48、72h时,收集各组细胞并提取总mRNA,RT-PCR法检测CX26、CX32、CX43基因表达的情况。通过染料传输实验,观察细胞间隙连接通讯功能的变化。结果SMMC-7721细胞和BEL-7404细胞在ATRA处理前没有CX26mRNA和CX32mRNA的表达,但均有CX43mRNA的表达。经ATRA处理后出现CX26mRNA和CX32mRNA的表达。经ATRA处理后SMMC-7721细胞间隙连接通讯功能增强,而且随ATRA作用时间延长,细胞间隙连接通讯功能增强越明显。BEL-7404细胞经ATRA处理24、48、72h后细胞间隙连接通讯功能没有明显的改变。结论全反式维甲酸能够在转录水平上调CX26、CX32基因在肝癌细胞SMMC-7721和BEL-7404中的表达。肝癌细胞SMMC-7721在CX26、CX32基因转录水平上调的同时伴有细胞间隙连接通讯功能的增强。肝癌细胞BEL-7404在CX26、CX32基因转录水平上调的同时不伴有细胞间隙连接通讯功能的增强。这说明两种细胞的间隙连接通讯功能调节机制不同。Objective To investigate the effect of all-trans retinoic acid (ATRA) on the expression of connexin 26, 32 and 43 genes and the alteration of gap junction communication function in human hepatocellular carcinoma cells. Methods Human hepatocellular carcinoma cell of the lines SMMC-7721 and BEL-7404 were cultured in normal medium and medium containing ATRA at a concentration of 10 -5 mol/L for 24, 48 and 72 hours respectively. RT-PCR procedure was adopted to detect the mRNA expression of CX 26, 32 and 43. Scrape-loading and dye transfer procedure was performed to examine the gap junction communication function. Results CX26 mRNA and CX32 mRNA were not expressed in the cell lines SMMC-7721, however, expression of CX26 mRNA and expression of CX32 mRNA were found 48 and 72 hours after being induction by ATRA respectively. CX26 mRNA and CX32 mRNA were not expressed in the cell lines BEL-7404, however, expression of CX26 mRNA and expression of CX32 mRNA were found 48 hours after induction by ATRA. Expression of CX43 mRNA was found in all cells, whether being induced by ATRA or not. Scrape-loading and dye transfer procedure showed that lucifer yellow was seen in only 1-2 lines by the delimited mark in the untreated SMMC-7721 cells and in 3-4 lines by the delimited mark in the SMMC-7721 cells treated by ATRA. But no dye transfer phenomenon was found in the BEL-7404 cells whether they were ATRA-treated or not. Conclusion ATRA is able to affect the expression of CX26 and CX32 in HCC cell lines SMMC-7721 and BEL-7404 by acting at the transcription level. Reinforcement of gap junction communication function is found in the SMMC-7721 cells and not in the BEL-7404 cells, which shows that ATRA modulates the gap junction intercellular communication, by acting in different mechanisms.

关 键 词：全反式维甲酸 肝癌细胞 连接蛋白基因 细胞间隙 连接通讯功能 

分 类 号：R735.7[医药卫生—肿瘤]