钙激活中性蛋白酶抑制剂对肝癌细胞生物学行为及Src基因表达的影响及意义

Effects of significance of calcium-activated neutral protease inhibitor on biological behavior and expression of Src gene in hepatocellular carcinoma cells

目的观察钙激活中性蛋白酶-2(Calpain-2)抑制剂ALLM对不同迁移能力肝癌细胞系侵袭迁移能力及Src基因表达的影响,并探索ALLM抑制肝癌的机制。方法常规细胞培养肝癌细胞系MHCC97-H、MHCC97-L、HepG2及正常人肝细胞HL-7702,经高浓度抑制剂ALLM处理后采用划痕实验、Transwell实验及western-bolt方法观察3株肝癌细胞及人正常肝细胞侵袭迁移能力及肝癌细胞中Src基因表达变化。结果与对照组比较,经ALLM处理的MHCC97-H、MHCC97-L的迁移距离、穿膜细胞的吸光度OD值差异具有统计学意义(P<0.05);转移性MHCC97-H、MHCC97-L肝癌细胞系Src蛋白的表达量显著下降(P<0.01);HepG2及正常人肝细胞HL-7702与转移性肝癌细胞迁移距离、穿膜细胞吸光度OD值及Src表达量比较差异具有统计学意义(P<0.05)。结论ALLM抑制Calpain-2的活性使肝癌细胞的迁移和侵袭能力显著下降,其机制可能与抑制原癌基因Src的表达和Src蛋白的活性有关。

Objective To investigate the effects of Calpain-2 inhibitor ALLM on invasion,migration ability and expression of Src gene in hepatocellular carcinoma cell lines with different migration abilities.Methods Conventional cell culture MHCC97-H,MHCC97-L,HepG2 and normal human liver cells HL-7702 cells.Observed the changes of three hepatocellular carcinoma cells and normal liver cells in the invasion,migration and expression of Src gene by the method of wound healing assay,transwell and Western-bolt,after treated by ALLM of high concentration.Results Compared with the control group,the migration distance of the ALLM-treated MHCC97-H,MHCC97-L,and the absorbance OD of the transmembrane cells were significantly difference(P<0.05).The expression of metastatic MHCC97-H,MHCC97-L liver cancer cell lines of Src protein weresignificantly decreased(P<0.01).Compared with HepG2 and normal human hepatocyte HL-7702,the migration distance,absorbance OD value of transmembrane cells,and Src expression was significantly difference in metastatic liver cancer cells(P<0.05).Conclusion ALLM inhibits the activity of Calpain-2 and significantly reduces the migration and invasion ability of liver cancer cells.The mechanism may be related to the inhibition of the expression of proto-oncogene Src and the activity of Src protein.