MST4表达对MHCC97H肝癌细胞细胞因子、ERK蛋白、p-ERK蛋白表达的影响及其意义

Effect of MST4 expression on the expression of cytokines, ERK protein and p-ERK protein in MHCC97H hepatoma cells and its significance

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 experimental absorbance value of the high expression group and the number of migration of the liver cancer cells in the Transwell experiment MHCC97H were higher than those of the empty group (P>0.05). In the white group and the low expression group (P<0.05), there was no significant difference between the MTT experimental absorbance value of the blank group and the low expression group and the number of MHCC97H hepatoma cell migration in the Transwell experiment (P>0.05). Conclusion:The high expression of MST4 in MHCC97H hepatoma cells mayactivate the expression of p-ERK protein,enhance the expression of cytokines, and in crease the value added and invasion of MHCC97H hepatoma cells.