肝细胞癌患者CNTN1检测的临床意义

Feasibility of contactin 1 as a novel prognostic marker for hepatocellular carcinoma

摘要:

检测肝细胞癌（HCC）患者肿瘤组织中接触蛋白1（CNTN1）的表达水平，及其对患者疾病预后价值的分析。方法采用实时荧光定量聚合酶链反应（qRT-PCR）和Western blot 检测25 例新鲜HCC 样本，以及邻近非肿瘤组织中CNTN1 mRNA 和蛋白的表达，同时采用免疫组织化学法检测135 例石蜡包埋HCC样本与邻近非肿瘤组织中CNTN1 的蛋白表达，统计分析其表达与HCC患者临床病理特征的关系，采用RNAi技术干扰肝癌细胞系HepG2 CNTN1的表达，对其进行肿瘤细胞侵袭和转移能力检测。结果25例新鲜HCC样本qRT-PCR和Western blot检测结果表明，与邻近非肿瘤组织相比，HCC 组织中CNTN1 mRNA 和蛋白表达均上调（ p<0.05）。免疫组织化学法结果证实62.22%（84/135）HCC 样本为CNTN1 阳性，高于邻近非肿瘤组织17.78%（24/135）（ p<0.05）。Kaplan–Meier 生存分析表明，CNTN1 阳性HCC 患者无疾病生存率和总生存率低于阴性患者。CNTN1 表达与HCC患者肿瘤转移、乙型肝炎病毒感染、淋巴结肿瘤转移分期及肿瘤直径相关（ p<0.05）。RNAi 实验表明，干扰CNTN1 表达后HepG2 细胞活力下降（ p<0.05）。多重变量分析表明，CNTN1 表达是HCC 患者无疾病生存期和总生存期的独立标志物（ p<0.05）。结论HCC 患者肿瘤组织中CNTN1 蛋白表达上调，且其表达与肿瘤恶性状态密切相关，参与肝细胞癌的侵袭和转移，可以作为HCC 患者疾病预后的独立标志物。

Abstract:

To evaluate the feasibility of contactin 1 (CNTN1) as a novel prognostic marker for epithelial hepatocellular carcinoma. Methods qRT-PCR and Western blot were performed to assess the mRNA and protein levels of CNTN1 in 25 matched fresh HCC specimens. The clinical and prognostic significance of CNTN1 in 135 cases of HCC was determined by immunohistochemistry. The correlations between CNTN1 expression and clinicopathological characteristics were analyzed. The expression of CNTN1 in HepG2 cells was interfered by RNAi technique, then the invasive and metastatic abilities of the HepG2 cells were determined. Results CNTN1 expression in the fresh HCC tissues was higher than that in the paracancerous tissues at both mRNA and protein levels ( p< 0.05). Notably, immunohistochemical results revealed that CNTN1 expression significantly increased in the HCC compared to the adjacent tissues (62.22% vs 17.78%,p < 0.05). The vitality of HepG2 cells was significantly decreased after RNA interference ( p< 0.05). Furthermore, positive CNTN1 expression was associated with tumor size, HBV infection, metastasis, and tumor-node-metastasis stage (p <0.05). Kaplan-Meier survival analysis showed that high CNTN1 was correlated with reduced overall survival(OS) rate (p < 0.05) and disease-free survival (DFS) rate ( p< 0.05). Multivariate analysis identified CNTN1 as an independent poor prognostic factor of OS and DFS in HCC patients ( p< 0.05). Conclusions Our resultssuggest that CNTN1 expression is up-regulated in HCC and closely correlated with the invasion and metasta-sis of HCC. It can be served as an independent unfavorable prognostic marker for HCC.