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研究力达霉素(LDM)与肿瘤坏死因子相关的凋亡诱导配体(TRAIL)联合作用对非小细胞肺癌细胞株H460细胞的增效作用及其作用机制。MTT法观察两药联合对H460细胞增殖的抑制作用。AnnexinV-FITC/PI双染、流式细胞术和Hoechst33342荧光染色检测两药联合对细胞凋亡的影响。Westernblotting检测凋亡通路中PARP、Caspase-3和Caspase-8分子的变化以及LDM对TRAIL受体DR4、DR5表达的影响。MTT结果显示,LDM与TRAIL的IC50值分别为4.603×10-10mol·L-1和915.3ng·mL-1,在TRAIL(50及100ng·mL-1)作用下LDM的IC50值分别为3.064×10-11和1.611×10-11mol·L-1。两药相互作用指数CDI<1。荧光显微镜下观察及流式细胞术检测两药联合细胞凋亡作用增强。联合用药组中Caspase-3和Caspase-8的激活作用明显增强。LDM能够增强TRAIL受体DR5的表达,不仅具有剂量依赖性,且作用时间越长表达量越高。研究结果提示,LDM对人非小细胞肺癌细胞株H460细胞具有生长抑制作用,可能通过诱导DR5表达上调而促进TRAIL诱导的肿瘤细胞凋亡,抑制细胞增殖,从而增加细胞对TRAIL的敏感性。
To investigate the synergistic effect and its mechanism of combined effect of Lidamycin (LDM) and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) on non-small cell lung cancer cell line H460. MTT assay was used to observe the inhibitory effect of the two drugs on the proliferation of H460 cells. AnnexinV-FITC / PI double staining, flow cytometry and Hoechst33342 staining were used to detect the effect of the two drugs on apoptosis. Western blotting was used to detect the changes of PARP, Caspase-3 and Caspase-8 in apoptotic pathway and the effect of LDM on the expression of TRAIL receptor DR4 and DR5. The results of MTT showed that the IC50 of LDM and TRAIL were 4.603 × 10-10mol·L-1 and 915.3ng · mL-1, respectively. The IC50 values of LDM with TRAIL (50 and 100ng · mL-1) were 3.064 × 10-11 and 1.611 × 10-11 mol·L-1. Two drug interaction index CDI <1. Fluorescence microscopy and flow cytometry to detect the combination of two drugs to enhance apoptosis. The activation of Caspase-3 and Caspase-8 in the combination group was significantly enhanced. LDM can enhance the expression of TRAIL receptor DR5, not only in a dose-dependent manner, but also in higher expression levels over longer periods of time. The results suggest that LDM can inhibit the growth of human non-small cell lung cancer cell line H460, which may promote the apoptosis of TRAIL-induced tumor cells by inducing the up-regulation of DR5 expression, inhibit the cell proliferation and increase the sensitivity of cells to TRAIL.