【摘要】 目的:通过检测HSP90β基因在白血病细胞株HL60及其耐药株HL60/ATRA中的表达差异,探讨HSP90β基因在ATRA相关多药耐药性发生中的作用。方法:采用ATRA浓度递增及间歇诱导法建立HL60/ATRA耐药细胞株;用MTT法和流式细胞检测技术(Flow cytometry,FCM),分别检测HL60和HL60/ATRA细胞的增殖及细胞周期;RT-PCR、Western blot法检测HSP90β基因在HL60及HL60/ATRA细胞中的表达;通过MTT法检测HL60和HL60/ATRA细胞对常用化疗药物(VCR、CTX、VP-16、ADM)敏感性。结果:成功构建了白血病细胞耐药株HL60/ATRA;在mRNA和蛋白两个水平,均检测到HSP90β在耐药株HL60/ATRA中的表达较HL60明显增高(P<0.01);药敏结果显示HL60/ATRA细胞对VCR、CTX、ADM、VP-16的耐药倍数分别为10.74、5.51、4.01、3.24。结论:ATRA能够诱导HL60细胞中HSP90β高表达,经ATRA诱导后的HL60/ATRA细胞对抗癌药物的敏感性显著降低、耐药性增高,这提示HSP90β可能在ATRA相关多药耐药性的发生过程中发挥着重要作用。更多还原

【Abstract】 Objective:To investigate the different expressions of heat shock protein 90β(HSP90β) mRNA and protein in all-trans retinoic acid(ATRA) sensitive leukemia cell line HL-60 and its resistant cell line HL60/ATRA in order to explore the effect of HSP90β on the ATRA multidrug resistance in leukemia.Methods:HL60/ATRA cell line was constructed by increasing ATRA concentration stepwisely and intermittently.The proliferation and sensitivity to anticancer drugs(VCR,CTX,VP-16,ADM) of HL60 and HL60/ATRA were tested by MTT assay.The cell cycle was analyzed using flow cytometry(FCM).The expression of HSP90β in HL60 cells and HL60/ATRA cells was detected with RT-PCR(reverse transcription PCR) and Western blot.Results:HL60/ATRA was successfully constructed.The transcription and protein expression of HSP90β in HL60/ATRA cells was higher than that of HL60 cells statistically(P<0.01).The drug resistant indexes of HL60/ATRA cells to VCR,CTX,VP-16 and ADM increased to 10.74,5.51,4.01 and 3.24,compared with HL60 cells respectively.Conclusion:ATRA could induce the high expression of HSP90 β in HL-60 cells.The sensitivity to anticancer drugs of HL60/ATRA induced by ATRA is decreased,but drug resistance is increased,which suggests that HSP90β may play an important role in the development of ATRA multidrug resistance.更多还原