【摘要】 目的观察小泛素样修饰蛋白-E2连接酶(UBE2I)在KRAS突变型结肠癌组织中的表达及其对KRAS突变型结肠癌细胞系增殖能力的影响。方法分析癌症基因组(TCGA)中86例已知KRAS突变类型的结肠癌样本中UBE2I差异表达水平及临床意义。构建短发夹RNA(shRNA)介导的UBE2I基因沉默KRAS突变型结肠癌细胞株。采用RT-PCR及Western blot检测结肠癌细胞中UBE2I mRNA与蛋白表达,CCK8增殖实验及细胞克隆实验分别检测细胞增殖及克隆能力变化。结果 UBE2I在KRAS突变型癌组织中表达明显上调(P <0.01),且其表达水平与KRAS突变型结肠癌患者临床分期及病理分级密切相关(病理分级P <0.01;临床分期P=0.02)。KRAS突变型结肠癌细胞HCT116、DLD1的UBE2I基因沉默后,克隆形成能力(P <0.01)及72 h后细胞增殖能力(P <0.01)均明显降低。结论 UBE2I在KRAS突变型结肠癌组织及细胞株中表达上调,沉默UBE2I基因可显著抑制肿瘤细胞的增殖及克隆能力。更多还原

【Abstract】 Objective To study the expression of small ubiquitin-KRAS mutation colorectal cancer and to explore its effects on the proliferation of KRAS mutation cancer cell.Methods The relationship between UBE2 I expression and external clinical information of KRAS mutation was identified by integrating The Cancer Genome Atlas(TCGA)database. Short-hairpin RNA directed against UBE2 I was inserted into the plasmid and then stable transfected cell lines. The proliferation and clone forming abilities were detected by cell counting kit-8 assay(CCK8)and colony formation. Results UBE2 I expression level was markedly up-regulated in KRAS mutation cancer samples when compared with that in paracancerous tissue(P <0.01)and KRAS wildtype cancer(P < 0.01). Strikingly,the expression of UBE2 I was significantly correlated with both pathologic grade and clinical stage(pathologic grade P < 0.01;clinical stage P = 0.02). Inhibition of UBE2 I by shRNA decreased the abilities of KRAS mutation cancer cell proliferation(both of HCT116 and DLD1 cell P <0.01 after 72 hours)and colony formation(both of HCT116 and DLD1 cell P < 0.01). Conclusion UBE2 I is over-expressed in KRAS mutation colorectal cancer. Knockdown of UBE2 I could inhibit the proliferation and cloning of KRAS mutation colorectal cancer cell.更多还原