白桦丹醌通过抑制PC9细胞迁移及PC9诱导的破骨细胞形成阻止肺癌骨转移的实验研究

Plumbagin Prevents Bone Metastasis of Lung Adenocarcinoma Cell Line PC9 by Inhibiting Migration and Osteolysis

目的:研究白桦丹醌在抗人类肺腺癌细胞系PC9骨转移中的作用。方法:采用左心室注射稳转荧光素酶的PC9细胞构建肺癌骨转移动物模型,研究白桦丹醌对PC9骨转移的抑制作用;采用SRB研究白桦丹醌对PC9的毒性作用,采用transwell迁移实验检测其对PC9体外迁移的抑制作用;采用PC9与Raw264.7共培养构建肺癌诱导破骨形成并用白桦丹醌处理,研究白桦丹醌对PC9诱导形成破骨细的抑制作用。结果:动物实验显示,心室注射PC9细胞40 d后,对照组5只裸鼠均出现明显转移,而用药组虽然也出现4只裸鼠转移,但用药组转移灶的光子值(2 759 200±2 670 290)显著小于对照组(8 732 800±6 191 628)(P<0.05)。SRB显示2μM白桦丹醌对PC9无明显杀伤作用,但可显著抑制PC9的迁移能力及显著抑制PC9诱导的破骨细胞形成,组间比较差异均有统计学意义(P<0.05)。结论:白桦丹醌可通过抑制PC9迁移及抑制PC9诱导的破骨细胞形成的方式抑制PC9骨转移,显示其作为抗肺癌骨转移药物的应用前景。

Objective：To investigate the effects of plumbagin on lung cancer bone metastasis. Method：Left ventricles injection of PC9 cells stably expressing luciferase was used to establish lung cancer bone metastasis model and the bone metastatic lesions were monitored by bioluminescent imaging and X-ray. The inhibiting effect against migration ability was detected by transwell assay and SRB test was applied to evaluated the inhibiting effects of proliferation. The coculture system of PC9 cells and Raw264.7 cells were preformed to establish the PC9 induced-osteoclast differentiation assay. The indicated concentrations of plumbagin were administrated to animals or PC9 cells.Result：In mouse model of bone metastasis, we demonstrated that plumbagin significantly repressed breast cancer cell metastasis and osteolysis. Mean photons emission of plumbagin group（2 759 200±2 670 290）were significantly lower than control group（8 732 800±6 191 628）（P〈0.05） 40 day after PC9 cells injection. In vitro experimentations showed 2μM Plumbagin significantly inhibited the migration of PC9 cells and osetoclasts formation induced by PC9 cells,but did not influence the proliferation of PC9 cells. Conclusion：Plumbagin has displayed the powerful anti bone metastasis effects against PC9 cell line and it might work via inhibiting cancer cell migration and cancer cell induced osteoclastogenesis,suggesting that Plumbagin could be a potential candidate in the treatment of breast cancer cell bone metastasis.