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目的 探讨p5 3抑制剂PFT α(p fiftythreeinhibitor ,PFT α)对热化疗诱导原代培养肠上皮细胞 (intestinalepithe lialcells ,IECs)凋亡及p5 3靶基因Bax和MDM2表达的影响。方法 原代培养IECs分为正常对照组、热化疗组和PFT α加热化疗组 ,运用顺铂联合温热 ( 4 3℃ )处理IECs 3 0min ,对比加入不同浓度的PFT α后 ,AnnexinV FITC /PI染色 ,流式细胞仪检测细胞凋亡。MTT法分析PFT α对热化疗杀伤DLD1的影响。Westernblot检测IECs的p5 3、Bax和MDM2蛋白表达。结果 热化疗导致IECs发生凋亡 ,Bax和MDM2蛋白表达明显高于对照组。 10、2 0、3 0、40 μmol/LPFT α作用于顺铂联合温热处理IECs后 ,细胞凋亡率下降且呈剂量依赖性 ,p5 3在IECs胞核 /胞浆表达比例下降 ,Bax和MDM2的表达随PFT α的剂量升高而逐渐下降。热化疗杀伤肿瘤细胞的效应并未受到低剂量PFT α影响。结论 PFT α对热化疗损伤肠上皮细胞具有保护作用 ,其机制可能与改变p5 3核转位和抑制促凋亡基因Bax和MDM2的表达相关 ,抑制p5 3可作为克服热化疗抗肿瘤治疗副效应的新策略。
Objective To investigate the effect of PFT inhibitor PFT α on the apoptosis of intestinalepithe lialcells (IECs) and the expression of p53 target genes Bax and MDM2 induced by thermochemotherapy in primary cultured human intestinal epithelial cells. Methods IECs of primary culture were divided into normal control group, thermochemotherapy group and PFT α heating chemotherapy group. IECs were treated with cisplatin and warm (43 ℃) for 30min. After adding different concentrations of PFTα, AnnexinV FITC / PI Staining and flow cytometry were used to detect apoptosis. The effect of PFT α on the killing of DLD1 by thermochemotherapy was analyzed by MTT method. Westernblot was used to detect the expression of p5 3, Bax and MDM2 in IECs. Results Thermal chemotherapy led to IECs apoptosis, Bax and MDM2 protein expression was significantly higher than the control group. After treated with 10, 20, 30, 40 μmol / L LPFA for 24 hours, the apoptotic rates of IECs were decreased and in a dose-dependent manner. The proportion of p5 3 in IECs decreased in nuclei and cytoplasm, The expression of MDM2 gradually decreased with the increase of PFT α dose. The effect of thermochemotherapy on killing tumor cells was not affected by low doses of PFTα. Conclusions PFT α may protect the intestinal epithelial cells from hyperthermia and its mechanism may be related to the change of nuclear translocation of p5 3 and the expression of pro-apoptotic genes Bax and MDM2. Inhibition of p5 3 can be used as a side effect to overcome the anti-tumor effect of thermochemotherapy New strategy.