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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 changes of CX26, CX32 and CX43 gene transcription in hepatocellular carcinoma cell lines SMMC-7721 and BEL-7404 induced by all-trans retinoic acid (ATRA) and the effect on cell gap junctional function. Methods The hepatocellular carcinoma cell lines SMMC-7721 and BEL-7404 were cultured with conventional medium and medium containing ATRA at a concentration of 10-5mol / L, and the cells were harvested at 24, 48 and 72 hours after drug-induced The total mRNA was extracted and the expression of CX26, CX32 and CX43 was detected by RT-PCR. Through the dye transmission experiment, observe the changes of cell gap junction communication function. Results SMMC-7721 cells and BEL-7404 cells did not express CX26 mRNA and CX32 mRNA before ATRA treatment, but all expressed CX43 mRNA. CX26 mRNA and CX32 mRNA expression after ATRA treatment. After ATRA treatment SMMC-7721 cell gap junctional intercellular communication function enhanced, and with the ATRA role of prolonged cell gap junctional communication function enhancement more obvious. BEL-7404 cells treated with ATRA 24,48,72 h after the cell gap junction communication function did not change significantly. Conclusion All-trans retinoic acid can upregulate the expression of CX26 and CX32 genes in hepatocellular carcinoma cells SMMC-7721 and BEL-7404 at the transcriptional level. Liver cancer cells SMMC-7721 in the CX26, CX32 gene transcription level increased accompanied by enhanced cell gap junctional communication. The upregulation of CX26 and CX32 gene transcription in hepatocellular carcinoma cell BEL-7404 is not accompanied by the enhancement of cell gap junctional communication. This shows that the gap between two kinds of cells to connect the communication function of different regulatory mechanisms.