肝门部胆管癌中CD133的表达及RNA干扰抑制对胆管癌细胞增殖的影响  

Expression of CD133 in cholangiocarcinoma cells and the effect of RNA interference on their proliferation

作　　者：张升涛[1] 刘斌[1] 常琦[1] 姬乐[1] 王加中[2] 

机构地区：[1]延安大学附属医院普通外科,延安医学硕士716000 [2]西安交通大学第二附属医院普外科,西安710003

出　　处：《医学研究生学报》2016年第11期1206-1210,共5页Journal of Medical Postgraduates

摘　　要：目的 CD133是肿瘤干细胞特征性的表面标记分子。文中探讨CD133在肝门部胆管癌组织中的表达以及抑制CD133表达对胆管癌细胞QBC939细胞增殖的影响。方法免疫组化法检测32例肝门部胆管癌组织、癌旁胆管组织中CD133的表达,分析CD133的表达水平与临床病理特征的关系。将CD133 siRNA转染至QBC939细胞内,通过Real-time PCR检测细胞转染后CD133 mRNA的表达,CCK-8法分别检测24、48、72 h后细胞转染情况。将实验细胞分为转染组(CD133 siRNA基因序列转染QBC939细胞组)、阴性对照组(阴性对照引物转染QBC939细胞组)以及空白对照组(无任何引物转染QBC939细胞组),每组实验重复3次。结果胆管癌组织中CD133阳性表达率62.5%(20/32),高于癌旁胆管组织中CD133阳性表达率28.1%(9/32),差异有统计学意义(P=0.0082)。CD133阳性表达可影响肿瘤的分化程度、TNM分期、淋巴结转移(P<0.05)。CD133 siRNA转染后,CD133 mRNA相对表达量低于空白对照组(0.437±0.014 vs 0.826±0.022,P<0.05)。转染24、48和72 h后,细胞的增殖活性受到明显抑制,与阴性对照组、空白对照组的差异均有统计学意义(P<0.05)。结论CD133在胆管癌组织中呈高表达,且影响肿瘤的分化、TNM分期、淋巴结转移。抑制CD133在胆管癌细胞QBC939的表达可抑制细胞增殖的活性。Objective CD133 is a surface marker of tumor stem cells. The aim of this study was to investigate the expression of CD133 in hilar cholangiocarcinoma and its effect on the proliferation of cholangiocarcinoma QBC939 cells. Methods The expression of CD133 was detected by immunohistochemistry in hilar cholangiocarcinoma and adjacent bile duct tissue in 32 patients,followed by analysis of the relationship between the expression level of CD133 and clinical pathological characteristics of the malignancy. CD133 siRNA was transfected into the QBC939 cells, the expression of CD133 mRNA in the cells determined by real-time PCR,and the status of transfection assessed after 24,48,and 72 hours by CCK-8.The cells were divided into a transfection group( CD133 siRNA gene transfected into QBC939 cells),a negative control group( negative control primers transfected into QBC939 cells),and a blank controlgroup( no primer transfected into QBC939 cells),and the experiment was repeated 3 times in each group. Results The positive expression of CD133 was significantly higher in cholangiocarcinoma than in the adjacent tissue( 62.5% [20 / 32]vs 28.1% [9 / 32],P =0.008 2) and it was closely related to the degree of tumor differentiation,TNM stage and lymph node metastasis( P<0.05). The relative expression of CD133 mRNA was markedly lower in the transfection than in the blank control group( 0.437±0.014 vs 0.826±0.022,P<0.05). At 24,48 and 72 hours after transfection,the proliferation activity of the cells was significantly inhibited as compared with that in the negative control or blank control group( P<0.05). Conclusion CD133 is highly expressed in cholangiocarcinoma and associated with tumor differentiation,TNM stage,and lymph node metastasis. The proliferation of cholangiocarcinoma QBC939 cells can be inhibited by suppressing the expression of CD133 in them.

关 键 词：胆管癌 CD133 RNA干扰 细胞增殖 

分 类 号：R657.4[医药卫生—外科学]