ADAM12在原发性肝癌侵袭性中的作用及机制研究

The role and mechanism of ADAM12 in invasiveness of primary hepatocellular carcinoma

目的观察ADAM12在原发性肝癌组织中的表达及与预后的关系,探讨ADAM12调控肝癌细胞的侵袭转移的作用及机制。方法采用免疫组化检测ADAM12和上皮-间充质转化(epithelial-mesenchymal transition,EMT)标志物在肝癌组织中的表达,统计分析对应原发性肝癌病人的预后情况。培养HepG2、MHCC97H肝癌细胞系,病毒转染,构建稳定的ADAM12沉默和过表达细胞。CCK8检测ADAM12对癌细胞增殖的影响,Transwell实验检测ADAM12对肝癌细胞侵袭能力的影响。Western Blot检测肝癌细胞中ADAM12和EMT标志物的表达改变。结果ADAM12表达水平越高,肝癌病人的预后可能越差,肿瘤≥3 cm可能性越大,分化程度越低,TNM分期越晚,肿瘤Ki67指数越高,且有统计学意义(P<0.05)。ADAM12的表达水平越高,肝癌增殖能力越大(P<0.05),抑制ADAM12的表达可降低肝癌细胞的增殖能力(P<0.05)。ADAM12的表达水平越高,肝癌侵袭潜能越大(P<0.05),抑制ADAM12的表达可抑制肝癌的侵袭(P<0.05),而过表达ADAM12可促进肝癌的侵袭(P<0.05)。ADAM12的表达水平与EMT标志物E-cadherin、N-cadherin、Vimentin的表达有关(P<0.05)。结论ADAM12通过调控肝癌EMT,增强HepG2、MHCC97H肝癌细胞增殖、侵袭能力。通过抑制ADAM12,可抑制肝癌细胞EMT,降低HepG2、MHCC97H肝癌细胞增殖、侵袭能力。

Objective To observe the expression of ADAM12 in primary hepatocellular carcinoma and its relationship with the prognosis,and to explore the role and mechanism of ADAM12 in regulating invasion and metastasis of hepatocellular carcinoma cells. Methods Immunohistochemistry was used to detect the expression of ADAM12 and Ki67 in hepatocellular carcinoma tissues,and the prognosis of patients with primary hepatocellular carcinoma was statistically analyzed.HepG2,MHCC97H hepatoma cell lines were cultured and transfected,and stable ADAM12-silenced and overexpressed cells were constructed.The effect of ADAM12 on the proliferation of hepatoma cells was detected by CCK8,and the effect on the invasiveness of hepatoma cells was detected by Transwell.Western-blot was used to detect the expression of ADAM12 and EMT markers in hepatoma cells. Results The higher expression of ADAM12 was associated with worse prognosis of patients with hepatocellular carcinoma,larger tumors, the lower the degree of differentiation,TNM stage,lymph node,the greater the possibility of vascular invasion,and higher tumor Ki67 index ( P <0.05). The higher expression level of ADAM12 was associated with the greater the proliferative capacity of HCC ( P < 0.05),and inhibition of ADAM12 expression reduced the proliferative capacity of HCC cells ( P <0.05).In addition,the higher expression level of ADAM12 was also associated with the greater the potential of invasion and metastasis of hepatocellular carcinoma ( P <0.05).Therefore,the inhibition of ADAM12 expression can inhibit the invasion and metastasis of hepatocellular carcinoma ( P < 0.05).Conversely,overexpression of ADAM12 can accelerate the invasion and metastasis of hepatocellular carcinoma ( P < 0.05 ).The expression level of ADAM12 was related to the expression of EMT markers E-cadherin,N-cadherin and vimentin ( P <0.05). Conclusion ADAM12 enhances the proliferation and invasion of HepG2 and MHCC97H hepatoma cells by regulating EMT.By inhibiting ADAM12,EMT of HCC cells and the proliferation as well as invasion of HepG2 and MHCC97H HCC cells can be suppressed.