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目的探讨PTEN对耐顺铂食管癌Ec9706/c DDP细胞增殖及P-gp表达的影响.方法采用顺铂浓度梯度增加的方法筛选食管癌耐药细胞株Ec9706/c DDP,采用流式细胞术检测PTEN转染后对Ec9706/c DDP细胞周期和凋亡的影响,采用Western blot检测PTEN转染后Ec9706/c DDP细胞P-gp表达改变。结果通过顺铂浓度梯度增加的方法成功诱导食管癌耐顺铂细胞株Ec9706/c DDP,与Ec9706/c DDP和Ec9706/c DDP+空载相比,转染野生型PTEN的Ec9706/c DDP细胞G1期细胞比例增加,S期细胞明显减少,差异有统计学意义(P<0.05)。Ec9706/c DDP细胞株在野生型PTEN转染48 h后细胞凋亡率显著高于转染无活性G129E和C124S组,差异有统计学意义(P<0.05)。Western blot结果显示:转染野生型PTEN的Ec9706/c DDP细胞P-gp蛋白的表达显著下调,而转染2种突变型的PTEN组没有明显改变。结论 PTEN在食管癌耐顺铂细胞株Ec9706/c DDP过表达能够显著抑制肿瘤细胞增殖,促进其凋亡,降低肿瘤耐药关键蛋白P-gp表达,从而逆转食管癌细胞的多药耐药。
Objective To investigate the effect of PTEN on the proliferation and P-gp expression of EC9706 / c DDP cell line.Methods Ec9706 / c DDP cells were screened by increasing the concentration of cisplatin.Flow cytometry PTEN transfected Ec9706 / c DDP cell cycle and apoptosis, Western blot was used to detect PTEN transfected Ec9706 / c DDP cells P-gp expression changes. Results EC9706 / c DDP cells were successfully induced by cisplatin concentration gradient. Compared with Ec9706 / c DDP and Ec9706 / c DDP + no-load cells, the transfection of wild-type PTEN Ec9706 / c DDP cells G1 The proportion of stage cells increased, S phase cells decreased significantly, the difference was statistically significant (P <0.05). The apoptotic rates of Ec9706 / c DDP cells transfected with wild type PTEN for 48 h were significantly higher than those transfected with inactive G129E and C124S groups (P <0.05). Western blot results showed that the expression of P-gp protein was significantly down-regulated in Ec9706 / c DDP cells transfected with wild-type PTEN, while the two mutant PTEN transfected groups did not change significantly. Conclusions Overexpression of PTEN in EC9706 / c cisplatin-resistant esophageal carcinoma cell lines can significantly inhibit tumor cell proliferation, promote apoptosis and reduce the expression of P-gp, a key protein of tumor resistance, thereby reversing the multidrug resistance of esophageal cancer cells.