Li Da-Wei, Dong Pin, Wang Fei, Chen Xin-Wei, Xu Cheng-Zhi, Zhou Liang
Department of Otolaryngology-Head and Neck Surgery, Affiliated Eye and ENT Hospital of Fudan University, Shanghai, China E-mail :
Asian Pac J Cancer Prev. 2013;14(8):4853-8. doi: 10.7314/apjcp.2013.14.8.4853.
To investigate whether hypoxia has an effect on regulation of multidrug resistance (MDR) to chemotherapeutic drugs in laryngeal carcinoma cells and explore the role of hypoxia-inducible factor-1α (HIF- 1α).
Laryngeal cancer cells were cultured under normoxic and hypoxic conditions. The sensitivity of the cells to multiple drugs and levels of apoptosis induced by paclitaxel were determined by MTT assay and annexin-V/propidium iodide staining analysis, respectively. HIF-1α expression was blocked by RNA interference. The expression of HIF-1α gene was detected by real-time quantitative RT-PCR and Western blotting. The value of fluorescence intensity of intracellular adriamycin accumulation and retention in cells was evaluated by flow cytometry.
The sensitivity to multiple chemotherapy agents and induction of apoptosis by paclitaxel could be reduced by hypoxia (P<0.05). A the same time, the adriamycin releasing index of cells was increased (P<0.05). However, resistance acquisition subject to hypoxia in vitro was suppressed by down-regulating HIF-1α expression.
HIF-1α could be considered as a key regulator for mediating hypoxia-induced MDR in laryngeal cancer cells via inhibition of drug-induced apoptosis and decrease in intracellular drug accumulation.
探讨缺氧对喉癌细胞化疗药物多药耐药（MDR）调控的影响，并探究缺氧诱导因子-1α（HIF-1α）的作用。
将喉癌细胞在常氧和缺氧条件下培养。分别通过MTT法和膜联蛋白V/碘化丙啶染色分析测定细胞对多种药物的敏感性以及紫杉醇诱导的凋亡水平。通过RNA干扰阻断HIF-1α表达。采用实时定量RT-PCR和蛋白质免疫印迹法检测HIF-1α基因的表达。通过流式细胞术评估细胞内阿霉素蓄积和保留的荧光强度值。
缺氧可降低细胞对多种化疗药物的敏感性以及紫杉醇诱导的凋亡（P<0.05）。同时，细胞的阿霉素释放指数升高（P<0.05）。然而，下调HIF-1α表达可抑制体外缺氧诱导的耐药性获得。
HIF-1α可被视为通过抑制药物诱导的凋亡和减少细胞内药物蓄积来介导喉癌细胞缺氧诱导的MDR的关键调节因子。
