摘要
目的观察细胞周期激酶亚单位1(CKS1)在肝细胞癌及正常肝组织中的表达以及其对人肝细胞癌细胞BEL-7405的增殖能力的影响。方法应用免疫组织化学检测65对手术肝细胞癌组织及癌旁正常组织CKS1的表达水平。应用Western印迹检测肝癌细胞系与正常肝脏细胞中CKS1表达量的差异。应用小干扰RNA(siRNA)技术转染肝细胞癌BEL-7405细胞,Western印迹检测CKS1 siRNA对CKS1蛋白的干扰效果,同时应用细胞计数试剂盒(CCK-8)和平板克隆增殖实验检测CKS1对BEL-7405细胞增殖能力的影响。结果免疫组织化学染色结果显示,CKS1在正常肝组织中表达量较肝细胞癌组织中表达量低,且差异有统计学意义(P<0.05)。Western印迹检测表明CKS1在7种肝细胞癌细胞系表达量显著高于正常肝细胞株,且CKS1的表达量在肝细胞癌BEL-7405细胞株显著高于其他肝癌细胞株。CCK-8法检测CKS1的siRNA转染BEL-7405细胞与阴性对照组和空白对照组相比明显抑制细胞增殖,在48、72和96 h A值分别降低了0.376、0.566、0.972(P<0.05);平板克隆增殖形成实验表明,CKS1的siRNA转染BEL-7405细胞后肿瘤细胞形成的克隆增殖集落明显少于阴性对照组和空白组,干扰组与空白、阴性对照组比较差异有统计学意义(P<0.05)。结论 CKS1在肝细胞癌组织中表达量显著高于正常肝组织,在肝癌细胞中表达量显著高于正常肝细胞,CKS1在肝细胞癌细胞BEL-7405的增殖中扮演了重要的角色,CKS1可能成为针对肝细胞癌基因治疗的新兴靶点。
Objective To investigate the expression of cyclin dependent kinase submit 1(CKS1)in hepatocellular carcinoma(HCC)and its effect on the proliferation of HCC cell line,BEL-7405.Methods The expression of CKS1 in 65 pairs of HCC tissues and normal tissues was measured by immunohistochemistry(IHC).Meanwhile,the expression of CKS1 in 7 different HCC cell lines and normal liver cell line was analyzed by western blot.Furthermore,a small interfering RNA(siRNA)targeting CKS1 was constructed and transfected into human HCC cells BEL-7405.The effects of CKS1 knockdown on cell proliferation and gene expression were assessed by CCK-8 assay,tablet cloning assay and western blot analysis,respectively.Results CKS1 expression was extremely higher in HCC tissue than that in normal liver tissue(P<0.05).Proliferation of siRNA-transfected BEl-7405 cells(siRNA-transfected group)was extremely inhibited comparing to that of negative control or blank group(P<0.05).Conclusion The expression of CKS1 in HCC tissues and cell lines was significantly higher than that normal liver tissue or cell line,respectively.Additionally,CKS1 plays an important role in proliferation of BEL-7405 HCC cell lines,which might be a novel anti-tumor target for HCC.
出处
《肝脏》
2016年第7期545-548,共4页
Chinese Hepatology
基金
国家自然基金项目(81472243)