HGF/cMet系统与肝癌细胞侵袭转移关系的实验研究

The correlation between the expression level of hepatocyte growth factor/scatter factor receptor cMet and invasiveness and metastasis in hepatocelluar carcinoma

目的:探讨HGF/cMet基因表达与肝癌细胞侵袭转移能力的关系。方法:采用荧光定量PCR和WesternBlot检测5株肝癌细胞系MHCC-1、HepG2、Huh7、SMCC-7721、Hep3B和永生化肝细胞株L02以及26份肝组织中cMetmRNA和蛋白质表达水平,同时采用免疫组化法检测26份肝组织中cMet蛋白质的表达。结果:6株肝癌细胞cMetmRNA分别为7.9×106,3.4×106,2.1×106,6.5×105,3.8×105和5.8×103(copies/ml),5株肝癌细胞系cMetmRNA显著高于L02细胞系(P<0.01),其中以转移能力较强的MHCC-1细胞系的cMetmRNA表达量最高;肝癌组织、癌旁组织及周围正常组织的cMetRNA分别为2.28×107、6.80×105、5.68×105,肝癌组织的cMet基因表达明显高于癌旁及周围正常组织(P<0.01);在肝癌组织中,T1、T2、T3/T4期及NO、N1/N2的cMetmRNA分别为4.88×104、6.98×105、1.72×106、4.67×105、8.73×105,cMet的表达与肿瘤的大小及其侵袭能力明显正相关;WesternBlot和免疫组化也显示不同肝细胞系蛋白质表达水平与cMetmRNA表达一致。结论:HGF/cMet系统参与了肝癌的形成,并且在肝癌细胞的侵袭和转移的过程中起重要作用。

Objective： To investigate the correlation between the expression level of cMet and invasiveness and metastasis in hepatocelluar carcinoma. Methods： The expression level of cMet at mRNA and protein level was assessed using real time PCR on five hepatocelluar carcinoma cell lines （ MHCC-1, HepG2, Huh7, Hep3B, SMCC-7721 ） and a immortalized hepatocyte cell line （L02）, 26 cases of hepatic tissues specimens including 12 hepatocelluar carcinoma tissues and 9 cases of surrounding normal tissue as well as 5 cases of normal hepatic tissues were also evaluated, Atthe same time we investigated the level of cMet protein on those 26 cases of hepatic tissues specimen by immunohistochemistry analysis. Results： The cMet mRNA copy number of 6 cell lines were 7. 9 × 10^6, 3, 4 × 10^6, 2. 1× 10^6, 6. 5 × 10^5, 3. 8 × 10^5 and 5.68 × 10^3 , respectively. Compared with L02 cell line, the expression level of cMet was found significantly more in five hepatocelluar carcinoma cell lines, among which MHCC-1 that had a potential of highly invaseness and metastasis was the most significantly （P 〈0. 01 ） ; The level of cMet mRNA of hepatocelluar carcinoma tissues was 2. 28 × 10^7, But there was 6. 80 × 10^5 and 5.8 × 10^5 in surrounding normal tissue and normal hepatic tissues ; The level of cMet of hepatocelluar carcinoma tissues was significantly higher than that of another two kinds （ P 〈 0. 01 ） ; c-Met mRNA copies were significantly correlated with the invasion; T1 versus T2, P 〈 0. 01 ; T1 versus T3/T4, P 〈 0. 01 ; T2 versus T3/T4, P 〈 0. 01 c-Met copy number in hepatocelluar carcinoma tissues of N1/N2 staged patients was significantly higher than NO cases （P 〈 0. 01 ）. There was a high correlation between the level of cMet and the size of tumor as well as lymph node metastasis. Expression levels of c-Met mRNA were also verified with immunohistochemistry analysis of c-Met protein expression in hepatocelluar carcinoma tissues and surrounding normal tissue as well as normal hepatic tissues. Conclusion： HGF/cMet participate in the cancerigenesis and may play an important role in the invasiness and mesatasis of hepatocelluar carcinoma.