论文部分内容阅读
目的:研究Plumbagin对人高转移大细胞肺癌L9981细胞系的体外抗肿瘤作用,并初步探讨其机制。方法:不同浓度Plumbagin处理L9981细胞系,采用MTT法观察对肿瘤细胞增殖的影响,确定药物IC50;采用流式细胞术检测对细胞系诱导凋亡的作用;采用Boyden小室侵袭实验检测对体外侵袭力的抑制作用。以IC50Plumbagin处理L9981细胞系,于处理后6、24、48h收获细胞,以实时定量PCR方法检测Bcl-2、Bax、VEGF和CYCD1等基因的mRNA表达变化。结果:MTT法显示,Plumbagin明显抑制L9981细胞系的细胞增殖(F=39.535,P=0.000),IC50为9.0μmol/L;Plum-bagin对L9981细胞系具有体外诱导凋亡作用(F=23.671,P=0.000),且明显抑制其体外侵袭能力。Bodyen小室侵袭实验检测对照组和Plumbagin组平均穿膜细胞数分别为228.17±55.12和9.83±3.87,差异有统计学意义,t=13.598,P=0.000。Plumbagin处理L9981细胞系后不同时间点检测结果显示,Bcl-2、VEGF和CYCD1基因表达均逐渐降低,bax基因表达逐渐增高,组间差异有统计学意义(F=13.520,P=0.000;F=15.778,P=0.000;F=10.163,P=0.000;F=18.635,P=0.000)。结论:Plumbagin对大细胞肺癌L9981细胞系具有明确的抗肿瘤作用。Plumbagin通过多种作用机制发挥其抑癌作用,显示了其成为抗大细胞肺癌药物的前景。
Objective: To study the anti-tumor effect of Plumbagin in human high metastatic large cell lung cancer L9981 cell line in vitro and its mechanism. Methods: L9981 cells were treated with different concentrations of Plumbagin. The effects of different concentrations of Plumbagin on cell proliferation were observed by MTT assay to determine the drug IC50. Flow cytometry was used to detect the effect on cell lines induced by apoptosis. Boyden chamber invasion assay was used to determine the in vitro invasiveness Inhibition. The L9981 cell line was treated with IC50Plumbagin, and the cells were harvested at 6,24,48h after treatment. The mRNA expression of Bcl-2, Bax, VEGF and CYCD1 were detected by real-time quantitative PCR. Results: MTT assay showed that Plumbagin significantly inhibited cell proliferation (F = 39.535, P = 0.000) and IC50 was 9.0 μmol / L in L9981 cell line. Plum-bagin induced apoptosis in L9981 cell line in vitro (F = 23.671, P = 0.000), and significantly inhibited its in vitro invasive ability. Bodyen chamber invasion assay detected that the average number of penetrating cells in the control and Plumbagin groups was 228.17 ± 55.12 and 9.83 ± 3.87, respectively, with a significant difference (t = 13.598, P = 0.000). The results of Plumbagin treatment of L9981 cell line at different time points showed that the expression of Bcl-2, VEGF and CYCD1 genes were gradually decreased and the expression of bax gene was gradually increased with a significant difference between the two groups (F = 13.520, P = 0.000; F = 15.778, P = 0.000; F = 10.163, P = 0.000; F = 18.635, P = 0.000). Conclusion: Plumbagin has definite anti-tumor effect on L9981 cell line of large cell lung cancer. Plumbagin exerts its anti-cancer effect through a variety of mechanisms of action, demonstrating its promise as an anti-large-cell lung cancer drug.