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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。
Objective To investigate the inhibitory effect of esophageal cancer related gene 2 (ECRG2) on malignant proliferation of EC9706 esophageal cancer cells and its mechanism. Methods ECRG2 eukaryotic expression plasmid (pcDNA3.1ECRG2) was constructed by DNA recombination technique. The ECRG2 protein was transfected into EC9706 cells and the growth of ECRG2 cells was observed. The changes of colony formation and soft agar colony formation were compared and analyzed. Effect of ECRG2 on malignant phenotype of EC9706. Further use of WesternBlot detection of p53, p21 changes. Results After transfected with ECRG2, the growth of EC9706 cells was inhibited and the proliferation rate of EC9706 cells was inhibited. The rate of platelet colony formation was 18% in experimental group and 55% in control group (P <0.05). The anchorage-independent ability of tumor cells decreased. The soft agarose colonies in the experimental group were small and formed in a small number (8 ± 1.88), which was significantly lower than that in the control group (14.3 ± 3.13) (P <0.05). Western Blot analysis showed that EC9706 cells transfected with ECRG2 expression accompanied by p53, p21 protein expression increased. CONCLUSION: Transfection of ECRG2 protein can partially reverse the malignant phenotype of EC8706 cells, which may be related to the p53 and p21 pathway.