机构地区:[1]郑州市第六人民医院肝病五科,郑州450000
出 处:《天津医科大学学报》2019年第2期124-127,共4页Journal of Tianjin Medical University
摘 要:目的:探讨MHCC97H肝癌细胞中丝氨酸/苏氨酸蛋白激酶4/(MST4)的表达与细胞因子、ERK蛋白、p-ERK蛋白表达的关系及其意义。方法:将MHCC97H肝癌细胞培养至对数生长期后,接种于96孔板上培养,分为空白组、高表达组(MST4转染高表达)、低表达组(MST4转染siRNA),采用Western-blot法检测各组共培养24 h后的MST4蛋白表达水平,采用ELISA法检测各组培养24 h后上清液中白细胞介素-2(IL-6)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、趋化因子-2(CCL2),采用Western-blot法检测各组的细胞外信号调节激酶(ERK)、磷酸化细胞外信号调节激酶(p-ERK)蛋白的表达,采用MTT实验检测3组肿瘤细胞的增殖能力,采用Transwell实验检测各组细胞的侵袭迁移能力。结果:高表达组的MST4蛋白表达显著高于空白组和低表达组(P<0.05);空白组和低表达组的MST4蛋白表达水平差异无统计学意义(P>0.05);高表达组的上清液中IL-6、IL-1β、TNF-α、CCL2水平均显著高于空白组和低表达组(P<0.05);空白组和低表达组的上清液中IL-6、IL-1β、TNF-α、CCL2水平差异无统计学意义(P>0.05);空白组、高表达组和低表达组的ERK蛋白表达差异无统计学意义(P>0.05);高表达组的pERK蛋白显著高于空白组和低表达组(P<0.05);空白组和低表达组的p-ERK蛋白差异无统计学意义(P>0.05);高表达组的MTT实验吸光度值、Transwell实验MHCC97H肝癌细胞迁移数目高于空白组和低表达组(P<0.05);空白组和低表达组的MTT实验吸光度值、Transwell实验MHCC97H肝癌细胞迁移数目差异无统计学意义(P>0.05)。结论:MHCC97H肝癌细胞中MST4高表达,将会激活p-ERK蛋白表达,从而提高细胞因子的表达,提高MHCC97H肝癌细胞的增殖、侵袭能力。Objective: To investigate the relationship between MST4 expression and cytokines, ERK protein and p-ERK protein expression in MHCC97H hepatoma cells and its significance. Methods:After the MHCC97H hepatoma cells were cultured at the logarithmic growth stage,they were inoculated on 96 orifice plates and divided into blank group-high expression group (MST4 transfection high expression) and low expression group (MST4 transfection siRNA). The ELISA method was used to detect interleukin -2 (IL-6), interleukin -1 beta (IL-1 beta) and tumor bad in the supernatant of each group after 24 h Death factor - alpha (TNF- alpha) and chemokine -2 (CCL2) were used to detect the expression of extracellular signal regulated kinase(ERK) and phosphorylated extracellular signal regulated kinase (p-ERK) protein in each group by Western-blot method. The increment ability of tumor cellsof three groups was detected by MTT test and the invasion of the cells was detected by Transwell test. Migration ability. Results:The expression of MST4 protein in the high expression group was significantly higher than that in the blank group and the low expression group (P<0.05). There was no significant difference in the expression of MST4 protein between the blank group and the low expression group (P>0.05).The levels of IL-6,IL-1 beta,TNF-α,and CCL2 in the supernatant of high expression group were significantly higher than those in the blank group and low expression group (P<0.05), and the differences in IL-6, IL-1 beta, TNF-α and CCL2 levels of the supernatant of the blank group and low expression group were not significant(P>0.05), but the negative expressions of the ERK protein in the blank group, the high expression group and the low expression group were poor. The p-ERK protein in the high expression group was significantly higher than that in the blank group and the low expression group(P<0.05), and the difference in the p-ERK protein of the blank group and the low expression group was not statistically significant (P>0.05);the MTT experimenta
关 键 词:MHCC97H肝癌细胞 丝氨酸/苏氨酸蛋白激酶4 增殖 侵袭迁移
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