磷脂酰肌醇蛋白聚糖-3的表达特征及其在肝癌诊断与鉴别诊断中的价值

Expression features of glypican-3 and its diagnostic and differential values in hepatocellular carcinoma

目的 研究磷脂酰肌醇蛋白聚糖-3（GPC-3）的表达特征及其在肝癌诊断与鉴别诊断中的临床价值.方法 制作鼠肝癌模型,并按病理组织检查结果分为正常组、肝细胞变性组（变性组）、癌前病变组（癌前组）和肝细胞癌组（癌变组）.以Western blot和逆转录-聚合酶链反应分别观察GPC-3蛋白质及mRNA的动态表达;以自身配对法收集术后肝癌组织,根据其组织学类型分为肝癌组、癌旁组、远癌组,以免疫组织化学法分析GPC-3表达与病理学特征的关系;以酶联免疫吸附法定量分析肝病患者外周血GPC-3表达水平,并评价其诊断效率.多个样本均数比较用单因素方差分析,组间GPC-3的表达及病理学特征比较用单因素秩和检验,血清GPC-3比较用秩和检验,率的比较采用x2检验或Fisher＇s exact分析,以受试者工作特征曲线下面积比较GPC-3诊断肝癌的敏感性、特异性及诊断效率.结果 在鼠肝癌形成过程中,正常、变性、癌前和癌变组GPC-3阳性率分别为0（0/6）、83.3%（15/18）、100.0%（9/9）和100.0%（9/9）.人肝癌组织GPC-3阳性呈棕黄色颗粒状染色,定位于胞质和细胞膜,肝癌、癌旁和远癌组的阳性率分别为80.6%、41.7%和0,肝癌组明显高于远癌组（x2=48.56,P＜0.01）和癌旁组（x2=11.455,P＜0.01）;癌旁组明显高于远癌组（x2=18.94,P＜0.01）.GPC-3表达与肿瘤分化程度和数目间未见明显相关,与瘤体大小有关（Z=2.941,P＜0.01）.肝病患者血清GPC-3异常主要见于肝癌（52.8%,65/123）,且在不同性别、年龄、甲胎蛋白水平、肿瘤数目、Child分级和肝外转移者间未见明显差异;但肝癌＜3.0cm组明显高于≥3.0cm组（x 2=6.318,P＜0.05）; HBsAg阳性组明显高于阴性组（x 2=23.362,P＜0.01）.GPC-3与甲胎蛋白（＞20 μg/L）联合诊断肝癌的敏感度、特异度、准确度、阳性预测值和阴性预测值分别为87.00%、79.66%、82.17%、69.03%和92.16%.结论 GPC-3表达与肝癌密切相关,其表达的检测有助于肝癌早期诊断和鉴别诊断.

Objective To investigate the expression features of glypican-3 （GPC-3）and its diagnostic and differential values in hepatocellular carcinoma （HCC）. Methods Rat hepatoma models were made and the dynamic expression features of GPC-3 protein and its gene were investigated by Western blotting and RT-PCR respectively. Liver specimens from 36 HCC patients were collected by self-control method and the expression and clinicopathological features of GPC-3 were analyzed by immunohistochemistry. Serum GPC-3 levels were quantitatively detected by ELISA and its efficiency for HCC diagnosis was evaluated in patients with liver diseases. Results The incidence of GPC-3 was 0% in control, 83.3% in degeneration, 100% in precanceration and 100% in canceration during dynamic formation of rat hepatoma, respectively. The positive GPC-3 was brown granule-like staining localized in membrane and cytoplasm in human HCC.The GPC-3 positive rates were 80.6% in HCC, 41.7% in surrounding tissues and none in distal tissues （P＜0.01）, respectively. No positive relationship presented between GPC-3 and differentiation grade or the number of minor except of rumor size （Z=2.941, P＜0.01）. The incidence of serum GPC-3 was 52.8% in HCC patients except of one patient with cirrhosis. No significant differences were found between GPC-3 and sex, age, AFP, mm or number, Child classification or extrahepatic metastasis except of rumor size （x2 = 6.318, P＜0.05） and HBV infection （x2 = 23.362,P＜0.01）. Combined detection of GPC-3 and AFP could rise up diagnosis of HCC. Conclusions GPC-3 expression closely associated with HCC and might be useful for early diagnosis of HCC.