ATRA促进DDP对肺腺癌细胞A549凋亡作用机制的研究

Study on the Mechanism about ATRA to Promote the Apoptosis of DDP on Lung Adenocarcinoma A549

探讨全反式维甲酸(all-trans retinoic acid,ATRA)对顺铂(cisplatin,DDP)抑制肺腺癌细胞株(A549)增殖的影响及其机制。采用四甲基偶氮唑盐比色法(MTT)、实时荧光定量逆转录聚合酶链反应(qRT-PCR)以及流式细胞术检测DDP、ATRA处理前后A549凋亡率及维甲酸受体-β(retinoic acid receptorβ,RARβ)mRNA、凋亡抑制蛋白Survivin mRNA的转录情况。结果表明DDP对A549有抑制作用,且呈剂量依赖性。ATRA小于0.4 mmol/L抑制作用不明显,ATRA达到0.4 mmol/L及更高浓度时,抑制作用显著,且呈剂量依赖性。qRT-PCR结果显示DDP组RARβmRNA及Survivin mRNA均降低。ATRA组RARβmRNA增加,而Survivin mRNA降低;联合用药组较空白组RARβmRNA降低,较DDP组RARβmRNA提高,而Survivin mRNA均降低。流式结果显示联合用药组较单独用药组凋亡率明显增高。ATRA上调RARβ并下调Survivin,提高A549化疗敏感性,强化DDP化疗作用。

To discuss the effect of all-trans retinoic acid（ATRA） on the cisplatin（DDP） induced lung adenocarcinoma cell line（A549） apoptosis and its mechanism.Using methyl thiazolyl tetrazolium salt assay（MTT）, real-time quantitative reverse transcription polymerase chain reaction（qRT-PCR） and flow cytometry to detect the A549 apoptosis rate,the transcription level of retinoic acid receptorβ（RARβ） mRNA and the transcription level of inhibitor of apoptosis protein Survivin mRNA.The results showed that DDP had inhibition effect on A549 in a dose-dependent way.When ATRA was less than 0.4μmol/L,the inhibition effect was not obvious,but when it up to 0.4μmol/L or higher concentrations,the inhibition effect was significantly in a dose-dependent way.The qRT-PCR results showed that the expression of RARβmRNA and Survivin mRNA all decreased in the DDP groups.In ATRA groups the expression of RARβmRNA increased,while the expression of Survivin mRNA decreased.In Combined treatment groups,the expression of RARβmRNA was lower than that in the Control groups but higher than that in the DDP groups.And when it comes to Survivin mRNA,it decreased both in Control groups and DDP groups.The flow cytometry results showed that the apoptosis rate in combined treatment groups were higher than that in single drug groups.In conclusion,the ATRA could promote the expression of RARβwhile inhibited the expression of Survivin.Through such means,ATRA could increase the chemosensitivity of A549 and strengthen the role of DDP chemotherapy.