Chinese Clinical Oncology
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WANG Xiaoxian, KONG Fanjun, LI Fei, ZHANG Shuo
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Abstract: 【Abstract】Objective To investigate the expression of miR-490-3p in adriamycin(ADM)resistant human breast cancer cell line MCF-7/ADM and its effect on the cell proliferation,apoptosis and resistance to ADM of MCF-7/ADM cell line. Methods Cultured wild-type MCF-7 human breast cancer cells and its resistant cells MCF-7/ADM were selected as research objectives. The realtime fluorescent quantitative PCR(qPCR)was used to compare the level of miR-490-3p in both cell lines. The recombinant plasmid pcDNA3.1(+)miR-490-3p to over-express miR-490-3p was transiently transfected into MCF-7/ADM cells(overexpression group)and then the transfection effect was verified by qPCR. Meanwhile,the pcDNA3.1(-)empty control group and blank control group were set up. MTT method was used to determine the effect of pcDNA3.1(+)miR-490-3p on the proliferation of each group. The inhibitory rate of ADM on the proliferation of MCF-7/ADM cells was determined, and the inhibition concentration (IC50) and the reversal factor were calculated to evaluate the sensitivity to ADM. The cells in each group were collected at 48 h post-transfection for the detection of apoptotic rate by flow cytometry with Annexin FITC/PI double staining. Meanwhile, the expression of resistance proteins including P-glycoprotein(P-gp),breast cancer resistance protein (BCRP)and multidrug resistance associated protein 1(MRP1)were tested by Western blotting,intracellular ADM concentration was detected by fluorescence photometer and Pgp activity by flow cytometry. Results The level of miR-490-3p in MCF-7/ADM cells was 0.24±0.07,significantly lower than 1.02±0.03 of wild type MCF-7 cells(P<005). The levels of miR-490-3p in the overexpression group were higher than those in the other two groups during 24-96 h post-transfection(P<005). The proliferation inhibition rates were(17.52 ±1.87)%,(31.67±2.79)%,(45.09±1.88)% and(61.82±2.52)% at 48,24,72 and 96 h post-transfection, all higher than those of empty control group(P<0.05). IC50 value of ADM for MCF-7/ADM cells was(11.27±2.34)μg/ml,lower than that of blank control group and empty control group(P<0.05). Moreover,reversal index was 3.35and 3.39fold of blank control group and empty control group for the overexpression group. Compared with the other two groups,there were increased apoptotic rates and intracellular drug concentration but decreased expression of P-gp,BCRP and MRP1 as well as P-gp activity in over-expression group (P<0.05).Conclusion Upregulation of miR-490-3p exhibits the reversal effect of resistance to ADM in MCF-7/ADM cell,possibly by reducing the resistance gene protein expression and inhibition of P-gp activity. Elevated level of miR-490-3p can inhibit proliferation and induce apoptosis.
WANG Xiaoxian, KONG Fanjun, LI Fei, ZHANG Shuo. Reversal effect of miR-490-3p on the resistance of MCF-7/ADM cells to adriamycin[J].Chinese Clinical Oncology, 2015, 20(9): 780-.
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http://manu65.magtech.com.cn/Jwk3_lczlxzz/EN/Y2015/V20/I9/780
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