热疗联合а-IFN逆转人肺腺癌细胞A549/CDDP耐药性的研究

Effect of Combiningа-IFN with Thermotherapy on Reversing the Drug Resistance of the Lung Adenocarcinoma Cell line A549/CDDP

目的探讨热疗联合α-IFN对人肺腺癌细胞A549/CDDP耐药逆转作用及可能机制。方法采用体外细胞培养技术,培养人肺腺癌细胞系A549及其耐药株A549/CDDP。设计1μg/ml、2μg/ml、4μg/ml浓度的CDDP,分别联合100U/ml、500U/ml、1000U/ml浓度а-IFN,在42℃热疗干预2h后,采用MTT法检测2种细胞对化疗药物CDDP的敏感性,采用流式细胞仪间接免疫荧光法,检测α-IFN、CDDP、热疗处理前后细胞中P-糖蛋白(P-gp)的表达及细胞内荧光药物CDDP的聚集量。结果热疗、а-IFN分别联用CDDP时,对A549/CDDP细胞的抑制率较单用CDDP时的抑制率高,差异有统计学意义(P<0.05)。热疗联合а-IFN与CDDP时,对A549/CDDP细胞的抑制率较热疗、а-IFN分别联用CDDP时明显升高(P<0.05)。热疗、а-IFN分别联合CDDP时,A549/CDDP细胞P-gp表达率较单用CDDP时下降,细胞内荧光强度增强,热疗联合α-IFN与CDDP,CDDP/A549P-gp表达率明显下降,细胞内荧光强度显著增强。结论а-IFN、热疗能部分逆转人肺腺癌细胞系A549/CDDP的耐药性;а-IFN联合热疗可提高逆转肺癌细胞耐药效应,其机制可能与抑制P-gp表达,增加细胞内化疗药物的积聚有关。

Objective To investigate the effect and the possible mechanism of combination of thermotherapy anda-IFN to the drug resistance of human lung adenocarcinoma cell line A549/CDDP. Methods Lung adenocarcinoma cell line A549 and drug resistant cell line A549/CDDP were cultured in vitro. After being treating by CDDP （1 μg/ml, 2 μg/ml and 4 μg/ml）, α-IFN （ 100 U/ml,500 U/ml and 1 000 U/ml） and thermotherapy（42℃ for 2 hours）, MTT method was used to detect the sensitivity of cell line A549 and the cell line Asg9/CDDP to CDDP. Flow cytometry detection was also used to determine the expression of P-gp and the deposition of CDDP in A549 and A549/CDDP cell line before and after the combined treatment. Results With α-IFN or thermotherapy ,the inhibition rate of CDDP to A549/CDDP cell line is higher than CDDP only used（ P 〈 0.05 ）. When treated with thermotherapy, α-IFN and CDDP, the inhibition rate of A.549/CDDP cell line is higher significantly than when treated by CDDP combined with α-IFN or themotherapy. When combined with thermotherapy and α- IFN respectively, the positive rate of p-gp expression decreased than CDDP only. When CDDP combined with thermotherapy and α-IFN, the positive rate of P-gp expression decreased further,And the dcpositiom of CDDP inside the cells increased further. Conclusion α-IFN and thermotherapy can reverse the drug resistance of lung ademocarcima cell line A549/CDDP in part, and the combination of α-IFN and themotherapy can increase this effect. The effect of combination of α-IFN and thermotherapy in reversing the drug resistance of A549/CDDP cell line may be correlated to the inhibition of P-gp expression and the increased deposition of the chemical drugs inside the cells.