食管癌相关基因2对EC9706细胞恶性增殖的抑制作用

目的探讨食管癌相关基因2(esophageal cancer related gene2,ECRG2)对EC9706食管癌细胞恶性增殖的抑制作用及其机制。方法利用DNA重组技术,构建ECRG2真核表达质粒(pcDNA3.1ECRG2),在食管癌EC9706细胞中转染并表达ECRG2蛋白,观察细胞的生长状态,比较平板集落和软琼脂克隆形成能力的改变,分析ECRG2对EC9706恶性表型的影响。进一步采用WesternBlot检测p53、p21的改变。结果转染表达ECRG2后,EC9706细胞的生长受到干扰,细胞的增殖速度受到抑制,实验组细胞平板集落形成率为18%,而对照组为55%,两者间差异显著(P<0.05);肿瘤细胞的停泊非依赖能力降低,实验组软琼脂克隆体积小且形成个数(8±1.88)明显少于对照组(14.3±3.13),差异显著(P<0.05)。WesternBlot分析表明,EC9706细胞转染表达ECRG2后伴随着p53、p21蛋白表达上升。结论转染表达ECRG2蛋白能够部分逆转食管癌EC8706细胞的恶性表型,其过程可能与p53、p21通路密切相关。

Inhibitory effects of esophageal cancer related gene 2 on proliferation of human esophageal cancer cell EC9706

Objective To investigate the inhibitory role of esophageal cancer related gene 2 (ECRG2) on proliferation of human esophageal cancer cell EC9706. Methods Recombinant plasmid pcDNA3.1-ECRG2 with ECRG2 open reading frame was constructed. The cells were transfected with either pcDNA3.1 or pcDNA3.1-ECRG2 using Lipofectamin~ TM 2000. The expression of ECRG2 protein was examined by Dot Blot analysis. The effects of ECRG2 on cell proliferation and malignant was analyzed by colony formation assay. The variation of P53 and P21 were detected in EC9706 cells with or without expression of ECRG2. Results The plasmid of pcDNA3.1-ECRG2 was successfully established. Colony formation activity of EC9706/pcDNA3.1-ECRG2 was 18% while that of the control cell was 55% in six well plate (P<0.05). The activity of anchorage-independent proliferation of EC9706/pcDNA3.1-ECRG2 was lower than that of EC9706/pcDNA3.1 in soft agar. After transfected with pcDNA3.1-ECRG2,the expression of P53 and P21 were higher than control. Conclusion ECRG2 can reduced the abilities of proliferation and of anchorage-independent proliferation of EC9706 cells which is through p53 pathway possibility.