腺病毒介导p53基因逆转乳腺癌耐药的实验研究

目的探讨腺病毒介导p53基因（Ad-p53）对人乳腺癌阿霉素（ADM）耐药细胞株MCF-7／ADM的耐药性及多药耐药基因1（MDR1）的影响。方法以Ad-p53转染MCF-7／ADM细胞株,流式细胞术检测p53蛋白变化;锥虫蓝活细胞拒染法观察细胞生长情况;MTT法观察MCF-7／ADM细胞对阿霉素耐药性的变化,实时荧光RT—PCR法检测MDR1 mRNA变化。结果流式细胞术观察到MCF-7／ADM细胞经MOI50的Ad-p53作用48h后,p53蛋白表达率由10．24％升高到36．20％;细胞抑制率为7．4％（P=0．003）;逆转MCF-7／ADM对ADM的耐药倍数11．6倍（P=0．001）;实时荧光RT-PCR检测结果显示MDR1 mRNA相对表达量由1．25±0．01下降至0．91±0．01（P=0．011）。结论腺病毒介导p53基因能抑制MCF-7／ADM细胞MDR1基因的表达,并能部分逆转MCF-7／ADM的耐药性,增加阿霉素化疗敏感性。

Experimental study of adenovirus-mediated p53 gene on the reversal of multidrug resistance in breast cancer

OBJECTIVE: To explore the effects of adenovirus-mediated wild-type p53 gene (Ad-p53) on the reversal of multidrug resistance (MDR) and multidrug resistance gene-1 (MDR1) in adriamycin (ADM) resistant human breast cancer cell. METHODS: Ad-p53 was transfected into the human breast cancer cells of the line MCF-7/ADM. p53 protein expression was detected by flow cytometry (FCM). The cell proliferation was assessed by trypan blue dye exclusion. The effects of Ad-p53 on ADM mediated drug resistance were observed by MTT assay. The expression levels of MDR1 mRNA were detected with real-time RT-PCR. RESULTS: The expression rate of p53 protein of the MCF-7/ADM cell was increased from 10.24% to 36.20% after transfected by MOI(50) Ad-p53 for 48 hours. The growth inhibition rate was 7.4 % (P = 0.003). The chemosensitivity of MCF-7/ADM to ADM increased by 11.6 times (P = 0.001). The result of real-time RT-PCR reveales that MDR1 mRNA was decreased from 1.25 +/- 0.01 to 0.91 +/- 0.01 (P = 0.011) after the MCF-7/ADM cells were transfected by Ad-p53. CONCLUSION: Ad-p53 can inhibit the expression of MDR1 gene and partially reverse the MDR of MCF-7/ADM cells, thus enhancing the chemosensitivity of human breast cancer cells to ADM.