低氧诱导因子-1α短发卡RNA逆转乳腺癌阿霉素耐药的研究

Reversal of adriamycin resistance induced by hypoxia inducible factor -1α short hairpin RNA in breast cancer

目的探讨低氧诱导因子-1α（HIF-1α）短发卡RNA（shRNA）沉默HIF-1α基因的表达对人耐阿霉素乳腺癌细胞MCF-7／ADR耐药能力的影响。方法应用HIF-1αshRNA重组载体质粒转染MCF-7／ADR细胞，荧光照微镜观察细胞转染过程，G418筛选稳定表达HIF-1αshRNA重组载体质粒的细胞，倒置显微镜观察低氧24、48h细胞生长情况，反转录-聚合酶链反应（RT—PCR）检测HIF-1α基因的表达情况，细胞计数试剂盒（CCK-8）实验鉴定细胞株的耐药性。结果荧光显微镜下叮见细胞内荧光显示，镜下可见低氧培养细胞典型凋亡，转染后的MCF-7／ADR细胞阿霉素敏感性增加（P〈0．05），对阿霉素半数抑制浓度（IC50）下降6．68倍。结论HIF—lotshRNA可以有效阻断MCF-7／ADR细胞HIF-1α的表达，抑制细胞生长，调控HIF-1α表达水平可能影响其细胞耐药特性，增加对化疗药物阿霉素的敏感性。

Objective To investigate the effect of hypoxia inducible factor （HIF） - 1α short hair- pin RNA （shRNA） on HIF - 1α gene expression and drug resistance of human breast cancer cells MCF -7/ADR cells. Methods HIF - 1α shRNA recombinant plasmid was transfected into MCF - 7/ ADR cells. G418 was used to observe the cell transfection. The growth of cells in 24 h and 48 h cells were observed under the inverted microscope. The expression of HIF - 1α was detected by reverse transcription polymerase chain reaction （ RT - PCR）. The drug resistance of ceils was detected by cell counting kit - 8 （ CCK - 8） assay. Results The cells showed a typical apoptosis under fluorescence microscope. The sensi- tivity to Adriamycin （ADR） of MCF -7/ADR cells was increased after transfection （P 〈 0. 05）, and the half maximal inhibitory concentration （IC50） was decreased by 6. 68 times. Conclusion HIF - 1α shRNA can effectively inhibit the expression of HIF - 1α and cell growth, which may affect the characteristics of drug resistance, and increase the sensitivity to chemotherapy drugs.