竹红菌乙素-光动力疗法对比杀伤经顺铂诱导耐药及非耐药肺癌细胞系的实验研究

Comparative Study on the Phototoxic Efficiency of HB-PDT to DDP-Resistant and Non-Resistant Lung Cancer Cell Lines

目的对比研究耐药及非耐药肺癌细胞株对竹红菌乙素-光动力疗法的反应性,以及耐药肿瘤细胞对竹红菌乙素-光动力疗法及化疗药物的反应性之间是否存在差别。方法以不同浓度的竹红菌乙素分别孵育人肺腺癌细胞系A549及经顺铂诱导耐药的多药耐药细胞系A549/顺铂,然后分别在铜蒸汽激光混合光饱和光剂量条件下进行照光处理,照光后继续孵育24 h,四唑盐比色法分别测定不同竹红菌乙素浓度、不同细胞的存活率,分别绘制竹红菌乙素对两种细胞存活率曲线并拟合曲线方程,根据方程求出竹红菌乙素对两种细胞的半数杀伤浓度。根据曲线、方程及半数杀伤浓度比较竹红菌乙素-光动力疗法杀伤经顺铂诱导耐药及非耐药细胞系的光毒效应有无差别;同时对比测定化疗药物顺铂对A549及A549/顺铂杀伤的半数杀伤浓度,比较竹红菌乙素-光动力疗法及化疗药物顺铂对耐药肿瘤细胞杀伤半数杀伤浓度间的差异。结果竹红菌乙素-光动力疗法对经顺铂诱导耐药及非耐药细胞系均具有很强的杀伤效应,竹红菌乙素对A549及A549/顺铂的半数杀伤浓度分别为33.82μg/L和34.19μg/L,竹红菌乙素-光动力疗法对两种细胞杀伤的半数杀伤浓度数据间统计学差异不显著(P>0.05)。顺铂对A549及A549/顺铂的半数杀伤浓度分别为1.06 mg/L和8.06 mg/L,两者之间统计学差异显著(P<0.01)。竹红菌乙素-光动力疗法和化疗药物顺铂对耐药肿瘤细胞杀伤效应间统计学差异显著(P<0.01)。结论竹红菌乙素-光动力疗法对于耐药及非耐药细胞的光毒效应无显著差别。竹红菌乙素-光动力疗法对耐药肿瘤细胞具有明显的光毒效应,其杀伤经顺铂耐药肿瘤细胞的效果明显优于已产生耐药的化疗药物。竹红菌乙素-光动力疗法有可能发展为对顺铂耐药肿瘤的有效治疗手段。

Objective To compare the reactivity of multidrug-resistant and non-drug-resistant lung cancer cell lines to HB-PDT, and to study whether there is difference between the reactivity of multidrug-resistant lung cancer cell line to HB-PDT and to chemotherapy. Methods Human lung adenocarcinoma cell line named A549 and multidrug-resistant cell line named A549/DDP were incubated with HB of different concentrations dissolved in DMEM, then irradiated by copper vapor laser with mixed wavelength light under saturated light dose. Cell survival rates at each concentration were measured by MTT assay after 24 hours＂ incubation, cell survival curves of the two kinds of cells were described and the equation of each survival curve was analyzed. IC50 of each cell was found from the equations. Finally the phototoxic efficiency of H/3-PDT to A549/DDP and A549 cell lines was analyzed according to the curves, equations and IC50 of the each kind of cells. At the same time, the IC50s of DDP for A549 and A549/DDP were also measured by MTT assay to find out if there were differences in the killing effect between DDP and HB-PDT to DDP-resistant lung cancer cell line. Results HP,-PDT had strong phototoxic efficiency to both DDP-resistant and non-resistant lung cancer cell lines. The IC50s of HB to A549 and A549/DDP were 33. 82 μg/L and 34. 19μg/L respectively. There was no significant difference between them（P〉0. 05）. On the other hand, the IC50s of DDP to A549 and A549/DDP were 1.06 mg/L and 8. 06 mg/L respectively（P〈 0. 01）. There was significant difference between the killing effects of HB-PDT and D to multidrug-resistant lung cancer cell line A549/DDP. Conclusion HB-PDT had strong phototoxic efficiency to DDP-resistant lung cancer cell line, the killing effect of HB-PDT was superior to that of DDP. There was no significant statistic difference in the phototoxic efficiency of HB-PDT to DDP-resistant and non-resistant lung cancer cell lines. HB-PDT probably can be developed to be an effective therapy to DDP-resistant lung cancer.