维甲酸联合维生素D_3对非小细胞肺癌耐药基因表达的影响及其意义的探讨

Effect and significance of Retinoic Acid and VD_3 reversing multidrug resistance in non-small-cell-lung-cancer ceiis

目的探讨全反式维甲酸(ATRA)联合维生素D3(VD3)逆转非小细胞肺癌细胞耐药的影响和意义。方法应用原代细胞培养、四甲基偶氮唑兰(MTT)比色、逆转录-聚合酶链反应(RT-PCR)等方法观察ATRA联合VD3对人肺癌原代细胞、肺腺癌A549细胞株化疗敏感性及多药耐药相关蛋白(MRP)、肺耐药蛋白(LRP)表达水平的影响。结果人肺癌原代细胞MRP、LRP高表达,其表达与肿瘤病理无明显相关,MRP表达阳性与ADM、VP-16、VCR的耐药相关,LRP表达阳性与DDP、CBP、ADM、VP-16、VCR的耐药相关;ATRA与VD3可以明显降低肺腺癌原代细胞及A549细胞株MRP、LRP的表达量,对肺鳞癌MRP、LRP的表达量无明显影响;可以明显提高肺癌化疗敏感性。结论MRP、LRP在非小细胞肺癌中高表达,可反映肿瘤细胞多药耐药性;ATRA或VD3可下调肺腺癌MRP、LRP基因的表达水平,二药合用有协同作用,可以提高肺腺癌的化疗敏感性;但对肺鳞癌MRP、LRP基因的表达水平无明显影响,可能还存在其他的提高肺鳞癌化疗敏感性的机制。

Objective ： To investigate the effect and significance of ATRA and VD3 reversing multidrug resistance in non-small-cell lung cancer cells. Methods ： Primary cell culture, MTT colorimetric assay and RT -PCR were applied to observe the different effects induced by ATRA or VD3 or ATRA and VD3 on lung cancer cells. Results ： The frequency of expression of MRP,LRP was 54.7% ,58.5% ,and the expression of MRP was correlated with drug resistance of ADM,VP-16, VCR, the expression of LRP was correlated with drug resistance of DDP, CBP, ADM, VP-16, VCR ; RT-PCR showed the significantly reduced expression of MRP and LRP in lung adenoeareinoma cells after ATRA and VD3 treatment. , but not in lung squamous carcinoma cells. And the IC50 were siguifieantly reduced after ATRA and VD3 treatment. And the effect,of ATRA and VD3 were synergistic. Conclusion： MRP,LRP were often expressed in NSCLC cells, and the expression of MRP,LRP was the guiding clue of multidrug resistance. ATRA and VD3 can increase the chemosensitivity by reduction of the expression of MRP,LRP in lung adenoeareinoma cells,but not in lung squamous carcinoma cells. It can increase the chemosensitivity by other mechanism in lung squamous carcinoma eeUs. The effect of ATRA and VD3 were synergistic.