Plumbagin对人高转移大细胞肺癌细胞系抑制作用的研究

目的:研究Plumbagin时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IC50 Plumbagin处理L9981细胞系,于处理后6、24、48 h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9.0 μmol/L;Plumbagin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.

Plumbagin对人类大细胞肺癌NL9980细胞系抑癌作用的研究

目的 研究Plumbagin对人类大细胞肺癌NL9980细胞系的抗肿瘤作用并探索其机制.方法 不同浓度Plumbagin处理NL9980细胞系,采用MTF法观察其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IC50 Plumbagin处理NL9980细胞系,于处理后6、24及48 h收获细胞,以实时定量PCR方法检测hcl-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NL9980细胞系的细胞增殖,IC50为7.5 μmol/L；Plumbagin对NL9980细胞系具有体外诱导凋亡作用,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161.59±47.32和26.58±9.07.Plumbagin处理NL9980细胞系后不同时间点检测结果显示,bcl-2、VEGF和CYCD1基因表达均逐渐减低,bax基因表达逐渐增高,组间差异有统计学意义(均P＜0.05).结论 Plumbagin对大细胞肺癌NL9980细胞系具有明确的抗肿瘤作用.Plumbagin可能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:The aim of this study was to investigate the effects of plumbagin (PL), a naphthoquinone derived from the medicinal plant plumbago zeylanica, on the invasion and migration of human breast cancer cells. Methods:Human breast cancer MDA-MB-231SArfp cells were treated with different concentrations of plum-bagin for 24 h. The effects of plumbagin on the migration and invasion were observed by a transwell method. The expressions of IL-1α, IL-1β, IL-6, IL-8, TGF-β, TNFα, MMP-2 and MMP-9 mRNA in M DA-MB-231SArfp cells were detected using Real-Time PCR. MDA-MB-231SArfp cells were treated with plumbagin at different concentrations for 45 minutes. The activation of STAT3 was detected by western blot. Following this analysis, STAT3 in MDA-MB-231SArfp cells was knocked out using specific siRNA. mRNA levels of IL-1α, TGF-β, MMP-2 and MMP-9 were then detected. Consequently, MDA-MB-231SArfp cells were injected intracardially into BALB/c nude mice to construct a breast cancer bone metastatic model. The mice were injected intra-peritoneally with plumbagin. Non-invasive in vivo monitoring, X-ray imaging and histological staining were performed to investigate the effects of plumbagin on the invasion and migration of breast cancer cells in vivo. Results: The in vitro results showed that plumbagin could suppress the migration and invasion of breast cancer cells and down-regulate mRNA expressions of IL-1α, TGF-β, MMP-2 and MMP-9. Western blotting demonstrated that plumbagin inhibited the activation of STAT3 signaling in MDA-MB-231SArfp cells. The inactivation of STAT3 was found to have an inhibitory effect on the expressions of IL-1α, TGF-β, MMP-2 and MMP-9. In vivo studies showed that plumbagin inhibited the metastasis of breast cancer cells and decreased osteolytic bone metastases, as well as the secretion of MMP-2 and MMP-9 by tumor cells at metastatic lesions. Conclusions:Plumbagin can suppress the invasion and migration of breast cancer cells via the inhibition of STAT3 signaling and by downregulation of IL-1α, TGF-β, MMP-2 and MMP-9.