FEN1过表达对肝细胞癌细胞生物学行为及患者预后的作用

Effects of FEN1 Overexpression on Biological Behaviors of Hepatocellular Carcinoma Cells and Prognosis of Patients

目的探讨FEN1在肝细胞癌中的表达及其在肝细胞癌发生发展中的作用。方法应用RTqPCR及免疫组织化学方法检测FEN1基因在52例肝细胞癌组织及相应癌旁组织中的表达。FEN1基因特异性siRNA转染肝癌细胞株HepG2并用Western blot检测转染效率。用MTT及流式细胞技术检测干扰FEN1后HepG2增殖及凋亡情况。Transwell实验检测HepG2迁移、侵袭能力改变。结果 RT-qPCR结果显示与癌旁正常组织相比较,FEN1高表达于肝细胞癌组织(P<0.01)。免疫组织化学与临床资料统计分析进一步证实FEN1的高表达与肿瘤分化程度(P=0.017)、肝内转移(P=0.046)、临床TNM分期(P=0.020)相关,而与患者性别、年龄、术前AFP值及肿瘤直径无相关性(P>0.05)。通过siRNA干扰肝癌细胞株HepG2中FEN1基因表达,MTT及流式细胞术检测表明干扰FEN1表达后肝癌细胞株HepG2增殖受到明显抑制(P<0.05),凋亡率明显增加(P<0.05)。Transwell实验发现干扰FEN1后可明显抑制HepG2迁移、侵袭能力(P<0.05)。结论 FEN1在肝细胞癌组织中高表达,高FEN1表达提示肿瘤分化程度低,易转移及预后差,干扰FEN1后抑制肝癌细胞增殖,诱导其凋亡,并降低肝癌细胞体外迁移和侵袭能力。FEN1基因可成为肝细胞癌诊断及治疗的一个新生物靶点。

Objective To investigate the expression level of FEN1 in hepatocellular carcinoma（HCC） cells and its role in the progression of HCC. Methods The expressions of FEN1 in 52 matched pairs of HCC tissues and corresponding normal tissues were measured by RT-qPCR and immunohistochemical staining. Specific FEN1 siRNA was transfected in HepG2 cells and the transfection efficiency was detected by Western blot. Moreover, cell proliferation was analyzed by MTT assay and cells apoptosis was determined by flow cytometry. Then, cell migration and invasion were detected by Transwell assays. Results FEN1 was overexpressed in HCC tissues in comparison with the corresponding normal tissues（P〈0.01）. FEN1 expression had positive correlations with differentiation degree（P=0.017）, intrahepatic metastasis（P=0.046） and TNM stage（P=0.020） of HCC tissues. However, as regards gender（P=0.731）, age（P=0.754）, level of AFP（P=0.076） and tumor size（P=0.100） no significant differences were observed between the groups. FEN1 expression in HepG2 cells could be downregulated by siRNA effectively. Moreover, the inhibition of FEN1 expression suppressed the proliferation and induced the apoptosis of HepG2 cells（P〈0.05）. Transwell migration assay suggested that the migration and invasion abilities of HepG2 cells were significantly inhibited when FEN1 was interfered（P〈0.05）. Conclusion FEN1 is abundantly expressed in HCC tissues, and abnormal FEN1 expression may associate with low differentiation grade, intrahepatic metastasis and poor patient prognosis. Downregulation of FEN1 expression reduced HCC cells proliferation and induced cells apoptosis, as well as, reduced HCC cells migration and invasion ability in vitro. FEN1 might be a novel target for hepatocellular carcinoma diagnosis and therapy.