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目的研究抑癌基因10号染色体缺失的磷酸酶和张力蛋白同源基因(PTEN)对耐顺铂食管癌细胞Ec9706/c DDP化疗敏感性的影响及其机制。方法建立耐药的食管癌细胞株Ec9706/c DDP,脂质体转染PTEN质粒及其功能缺陷的突变质粒PEGFP-C1-G129E和PEGFP-C1-C124S,Wst-1检测食管癌细胞Ec9706/c DDP在含顺铂培养基中的增殖情况,克隆形成实验检测其增殖能力,流式细胞术检测其凋亡,荧光定量聚合酶链反应(Q-PCR)检测多重耐药基因1(MDR1)mRNA的表达水平。结果与完全空白组、转染空质粒组、PEGFP-C1-G129E组和PEGFP-C1-C124S组相比,转染PTEN的Ec9706/c DDP细胞增殖明显减少(P<0.01);转染PTEN组Ec9706/c DDP克隆形成率减少;转染PTEN使得Ec9706/c DDP凋亡增加(P均<0.01);Q-PCR结果显示,PTEN明显抑制MDR1 mRNA水平的表达(P均<0.01),并且这种抑制作用依赖于129和124这两个位点。结论 PTEN可促进耐药食管癌细胞的化疗敏感性,并通过抑制MDR1基因的表达水平促进肿瘤细胞对顺铂的敏感性。
Objective To investigate the effect and mechanism of PTEN on the chemosensitivity of cisplatin-resistant esophageal cancer cells Ec9706 / c DDP. Methods Ec9706 / c DDP cells were transfected with esophageal cancer cell lines Ec9706 / c, PEGFP-C1-G129E and PEGFP-C1-C124S which were transfected with PTEN plasmid and their defective plasmids. DDP proliferation in cisplatin-containing medium, clone formation assay to detect its proliferation ability, flow cytometry to detect apoptosis, quantitative polymerase chain reaction (Q-PCR) detection of multidrug resistance gene 1 (MDR1) mRNA The level of expression. Results The proliferation of Ec9706 / c DDP cells transfected with PTEN was significantly decreased (P <0.01) compared with those of blank control group, empty plasmid transfected group, PEGFP-C1-G129E group and PEGFP-C1-C124S group Ec9706 / c DDP clone formation rate decreased; PTEN transfected Ec9706 / c DDP apoptosis increased (P <0.01); Q-PCR results showed that PTEN significantly inhibited MDR1 mRNA expression (P <0.01), and this The inhibitory effect depends on the two sites 129 and 124. Conclusion PTEN can promote the chemosensitivity of esophageal cancer cells and promote the sensitivity of tumor cells to cisplatin by inhibiting the expression of MDR1 gene.