不同转移潜能的肝细胞肝癌细胞中EMS1/cortactin的表达与定位

目的:研究不同转移潜能的肝细胞肝癌(hepatocellular carcinoma,HCC)细胞中株中EMS1/cortactin的表达与定位,探讨EMS1/cortactin与HCC细胞生物学行为的相关意义。方法:在不同生物学特性的HCC细胞株较高转移潜能的MHCC-97H(97H),惰性潜能的SMMC-7721(7721)和HepG2]中采用RT-QPCR、Western Blot和免疫荧光方法检测EMS1基因和cortactin蛋白的表达,在共聚焦显微镜下观察cortactin蛋白在细胞中的定位。结果:97H、7721、HepG2细胞中EMS1 mRNA水平分别为正常肝细胞株L02的23.5倍、15.3倍和10.5倍;cortactin水平分别为L02的3.6倍、2.7倍和2.6倍(均P<0.05)。HCC细胞株中cortactin蛋白表达高于L02。97H细胞中EMS1/cortactin的表达均高于其他两株HCC细胞。各型HCC细胞株中cortactin蛋白聚集在细胞膜下的皮质区。L02细胞中cortactin蛋白弥散分布在胞质中。结论:HCC细胞中EMS1基因和cortactin蛋白高表达在癌细胞膜下的皮质区,与癌转移潜能相关。

Expression and location of EMS1/cortactin in different hepatocellular carcinoma cell lines

Objective: To investigate the location,expression of EMS1/cortactin in hepatocellular carcinoma(HCC) cell lines.Methods: In HCC cell lines(97H,7721 and HepG2),expression of EMS1/cortactin were detected with RT-QPCR,Western Blot assay respectively.The location of cortactin was detected with immunofluorescence assay by confocal microscopy.L02 was used as control.Results: As compared with L02,the levels of EMS1 gene and cortactin protein in 97H,7721 and HepG2 were 23.5(relative fold units),15.3,10.5,3.6,2.7,2.6 in 97H,7721 and HepG2 and L02,respectively.Cortactin accumulated in the the cell-substratum contact sites to a high extent in HCC cell line,especially in 97H cell line(P<0.05).Conclusion: EMS1/Cortactin showed high expression in HCC cell lines.Cortactin accumulated in the the cell-substratum contact sites to a high extent in HCC.Cortactin might contribute to the invasive potential of these tumor cells.