Liu Ying-bin, Xu Bin, Wang Jian-wei, Fang He-qing, Li Jiang-tao, Li Hai-jun, Tang Zhe, Qian Hao-ran, Feng Xue-dong, Peng Shu-you
Second Affiliated Hospital, Zhejiang University Medical School, Hangzhou 310009, China.
Zhonghua Yi Xue Za Zhi. 2005 Jun 1;85(20):1414-8.
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.
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.
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.
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.
研究全反式维甲酸（ATRA）对人肝癌细胞中连接蛋白26、32和43基因表达及缝隙连接通讯功能改变的影响。
将人肝癌细胞系SMMC - 7721和BEL - 7404分别在正常培养基和含10(-5) mol/L ATRA的培养基中培养24、48和72小时。采用RT - PCR法检测CX 26、32和43的mRNA表达。进行刮擦加载和染料转移实验以检测缝隙连接通讯功能。
在SMMC - 7721细胞系中未检测到CX26 mRNA和CX32 mRNA表达，但经ATRA诱导48小时和72小时后分别检测到CX26 mRNA和CX32 mRNA表达。在BEL - 7404细胞系中未检测到CX26 mRNA和CX32 mRNA表达，但经ATRA诱导48小时后检测到CX26 mRNA和CX32 mRNA表达。无论是否经ATRA诱导，在所有细胞中均检测到CX43 mRNA表达。刮擦加载和染料转移实验显示，未处理的SMMC - 7721细胞中，荧光素黄仅在1 - 2个界限标记的细胞中可见；经ATRA处理的SMMC - 7721细胞中，荧光素黄在3 - 4个界限标记的细胞中可见。但无论是否经ATRA处理，在BEL - 7404细胞中均未发现染料转移现象。
ATRA能够通过作用于转录水平影响肝癌细胞系SMMC - 7721和BEL - 7404中CX26和CX32的表达。在SMMC - 7721细胞中发现缝隙连接通讯功能增强，而在BEL - 7404细胞中未发现，这表明ATRA通过不同机制调节缝隙连接细胞间通讯。
