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目的探讨Shc1在肝癌组织中的表达情况及其对人肝癌SMMC-7721细胞增殖、迁移能力的影响。方法采用实时定量PCR、蛋白质印迹法、免疫组织化学法检测33例肝癌及其对应的癌旁组织中Shc1的表达情况,并分析Shc1蛋白表达量与临床特征间的关系;培养肝癌SMMC-7721细胞,以siRNA技术干扰Shc1的表达,采用MTT法和细胞划痕试验分别检测干扰Shc1对细胞增殖活力和迁移能力的影响。结果在33对样本中,肝癌组织中Shc1的表达高于癌旁组织(P<0.05);Shc1表达于肝癌细胞的胞质中。无肝硬化的患者Shc1阳性率(100%)高于伴肝硬化者(54.2%),术前Child-Pugh分级A级的患者Shc1阳性率(75.9%)高于B级或C级的患者(0%),术前血AFP阳性患者Shc1阳性率(79.2%)高于AFP阴性患者(33.3%),术后病理EdmondsonⅢ、Ⅳ级患者Shc1阳性率(84.2%)高于Ⅰ、Ⅱ级的患者(42.9%),差异均有统计学意义(P<0.05)。干扰Shc1表达后,随时间延长,SMMC-7721细胞增殖、迁移明显受到抑制(P<0.05)。结论 Shc1在肝癌组织中高表达,且Shc1的阳性率与有无肝硬化、Child-Pugh分级、术前AFP、病理Edmondson分级相关;干扰Shc1的表达可抑制肝癌SMMC-7721细胞的增殖、迁移能力。
Objective To investigate the expression of Shc1 in hepatocellular carcinoma and its effect on the proliferation and migration of human hepatoma SMMC-7721 cells. Methods The expression of Shc1 was detected by real-time PCR, Western blotting and immunohistochemistry in 33 HCC tissues and their corresponding paracancerous tissues. The relationship between Shc1 protein expression and clinical features was analyzed. The hepatoma SMMC-7721 cells were cultured SiRNA was used to interfere with the expression of Shc1. MTT assay and cell scratch assay were used to detect the effect of Shc1 on cell proliferation and migration. Results In 33 pairs of samples, the expression of Shc1 in HCC tissues was higher than that in adjacent tissues (P <0.05). Shc1 was expressed in the cytoplasm of HCC cells. The positive rate of Shc1 in patients without cirrhosis (100%) was higher than that in patients with cirrhosis (54.2%). The positive rate of Shc1 in patients with Child-Pugh grade A before operation (75.9%) was higher than those in patients with Grade B or C 0%). The positive rate of Shc1 in preoperative blood AFP positive patients (79.2%) was higher than that in AFP negative patients (33.3%). The positive rate of Shc1 in patients with Edmondson Ⅲ and Ⅳ grade was 84.2% (42.9%), the differences were statistically significant (P <0.05). After Shc1 expression was disturbed, the proliferation and migration of SMMC-7721 cells were significantly inhibited with time (P <0.05). Conclusion Shc1 is highly expressed in HCC and the positive rate of Shc1 is related to the presence or absence of cirrhosis, Child-Pugh classification, preoperative AFP and pathological Edmondson classification. Interfere with Shc1 expression can inhibit the proliferation and migration of HCC SMMC-7721 cells.