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肝细胞癌中Nodal的表达及其与血管生成和上皮间质转化的关系

Expression of Nodal in hepatocellular carcinoma and its relationship with angiogenesis and epithelial-mesenchymal transition

摘要:

目的 探讨Nodal在肝细胞癌中表达情况,及其与血管生成和上皮间质转化(EMT)的关系.方法 2006年9月到2010年6月,收集16例冷冻肝癌组织和癌旁组织,应用实时荧光定量PCR(qPCR)和Western blot检测Nodal的表达;收集10例正常肝组织、96例肝癌组织和癌旁组织,采用免疫组化染色检测Nodal的表达,分析Nodal表达与肝癌临床病理特征及生存预后之间的关系.同时分析96例肝细胞癌患者癌组织Nodal、CD34和Vimentin的表达,探讨Nodal的表达与血管生成、EMT间的关系.结果 免疫组化染色显示Nodal主要在细胞质中表达.肝癌组织中高表达Nodal的比率为72.9% (70/96),显著高于癌旁组织(8.3%)和正常组织(0) (x2 =83.001、24.470,P<0.001).qPCR和Western blot检测结果显示,肝癌组织中Nodal表达水平明显高于癌旁组织和正常肝组织(P<0.05).Nodal高表达与肿瘤最大径(x2=15.318,P=0.000)、甲胎蛋白(x2=3.850,P=0.049)、吲哚氰绿15 min潴留率(x2=6.590,P=0.010)和侵袭转移(x2=17.824,P =0.000)有关.免疫组化染色检查发现,肝癌组织中Nodal的表达与微血管密度(MVD)有关(t=3.070,P=0.006),与Vimentin的表达无关(r=0.198,P=0.053).生存分析发现,高表达Nodal的患者术后积累生存率明显小于低表达者(x2 =487.053,P<0.001).多因素分析表明,Nodal高表达是影响肝癌患者术后生存的独立危险因素(RR =2.757,95% CI:1.450~5.240,P=0.002).结论 Nodal不参与肝癌EMT,但可促进肝癌血管形成,可作为判断肝癌患者生存预后的指标之一.

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abstracts:

Objective To detect the expression of Nodal in hepatocellular carcinoma (HCC),and explore its relationship with angiogenesis and epithelial-mesenchymal transition (EMT).Methods From September 2006 to June 2010,the 16 self-paired frozen HCC specimens were collected and the expression of Nodal was detected by qPCR and Western blot.The 10 normal liver tissues and 96 cases of HCC tumor and paracarcinomatous tissues were collected.The expression of Nodal and relationship among Nodal,clinicopathological characteristics of HCC and patients' prognosis were detected and analyzed using immunohistochemistry.The expressions of Nodal,Vimentin and CD34 in 96 HCC tumor tissues were detected by immunohistochemistry,and then judgment relationship between the expression of Nodal,EMT and angiogenesis.Results Immunohistochemistry showed that Nodal mainly expressed in the cytoplasm.The high expression rate of Nodal in HCC tumor tissues was 72.9% (70/96),which was remarkably higher than that in paracarcinomatous tissues (8.3%) and normal liver tissues (0) (x2 =83.001 and 24.470,both P<0.001).qPCR and Western blot analysis showed that the expression level of Nodal in HCC was significantly higher than that in paracarcinomatous and normal tissues (P < 0.05).The high expression of Nodal in HCC was correlated with tumor size (x2 =15.318,P =0.000),alpha-fetoprotein (x2 =3.850,P =0.049),indocyanine green retention rate at 15 minutes (x2 =6.590,P =0.010),and invasion and metastasis (x2 =17.824,P =0.000).High expression of Nodal was positively correlated with high microvascular density in HCC (t =3.070,P =0.006),but not with Vimentin (r =0.198,P =0.053).Survival analysis showed that accumulated survival rate of patients with high expression of Nodal was significantly less than that the low expression (x2 =487.053,P < 0.001).The Cox multivariate analysis demonstrated that high expression of Nodal was independent risk factors for cumulative survival in patients with hepatocellular carcinoma after a curative resection (RR =2.757,95% CI:1.450-5.240,P =0.002).Conclusions Nodal does not participate in EMT of HCC,but can promote angiogenesis,and it could be used as a predictor of poor prognosis.

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作者: 陈静 [1] 李建生 [1] 刘文斌 [1] 荚卫东 [1] 许戈良 [1] 马金良 [1]
第一作者: 陈静
期刊: 《中华外科杂志》2014年52卷3期 188-192页 MEDLINEISTICPKUCSCD
栏目名称: 论著
DOI: 10.3760/cm.j.issn.0529-5815.2014.03.008
发布时间: 2014-04-01
基金项目:
国家自然科学基金资助项目 安徽省科技攻关资助项目 安徽省自然科学基金资助项目 安徽省高校自然科学研究资助项目
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