Yu Shao-hong, Yan Lü-nan, Zhang Yan, Gou Xing-hua, Han Lei, Chen Yong-bing
Department of Hepatobiliary Surgery, Chongqing Fuling Center Hospital, Fuling 400800, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2005 Jul;36(4):471-4.
To conduct an in vitro study on the effect of recombinant adenovirus microsphere encapusulated antisense MRP (as-mrp) for use in the gene therapy to overcome drug resistance in hepatocellular carcinoma.
Recombinant adenovirus microsphere encapusulated as-mrp was transfected into hepatocellular carcinoma multidrug resistance cells HepG2/ADM, the fluorescence intensity of transfected cells were observed at 48 hours and 120 hours after transfection. in vitro drug sensitivity was measured by MTT assay; the resistant index of andromycin resistant variants was determined by drawing the cell dosage reaction curves. The levels of MRP mRNA expression were detected by RT-PCR and the ratio of MRP mRNA/beta-actin was detected. Intracelluar rubidomycin (DNR) concertration was examined by flow cytometry (FCM).
More than 90% of the HepG2/ADM cells could be transfected when microspheres being 10 mg. Adv microsphere inhibited the expression of mRNA in HepG2/ADM and enhanced the sensitivity of HepG2/ADM to chemotherapeutic drug.
Recombinant adenovirus microsphere encapusulated as-mrp could effectively reverse HepG2/ADM cells, which would provide an experimental basis for the methods of reversing the multidrug resistance in human hepatocellular carcinoma.
进行一项体外研究，探讨重组腺病毒微球包裹反义多药耐药相关蛋白（as-mrp）在肝癌基因治疗中克服耐药性的作用。
将重组腺病毒微球包裹的as-mrp转染至肝癌多药耐药细胞HepG2/ADM，于转染后48小时和120小时观察转染细胞的荧光强度。采用MTT法检测体外药物敏感性；通过绘制细胞剂量反应曲线测定阿霉素耐药变异体的耐药指数。采用RT-PCR检测MRP mRNA表达水平，并检测MRP mRNA与β-肌动蛋白的比值。通过流式细胞术（FCM）检测细胞内柔红霉素（DNR）浓度。
当微球为10mg时，超过90%的HepG2/ADM细胞可被转染。腺病毒微球抑制了HepG2/ADM中mRNA的表达，并增强了HepG2/ADM对化疗药物的敏感性。
重组腺病毒微球包裹as-mrp可有效逆转HepG2/ADM细胞，为逆转人肝癌多药耐药的方法提供实验依据。
