mTOR抑制剂雷帕霉素对黑色素瘤肝转移的影响

Effect of mTOR inhibitor rapamycin on liver metastasis of melanoma

目的:评价mTOR抑制剂雷帕霉素对黑色素瘤转移的影响,为其临床应用奠定基础。方法:通过脾内注射建立黑色素瘤肝转移模型,测试雷帕霉素对黑色素瘤转移的影响,并通过HE染色作病理验证。在体外培养体系中,通过MTT实验、划痕实验、Transwell实验、流式实验检测药物对黑色素瘤细胞增殖、迁移、侵袭、凋亡能力的影响。结果:在肝转移模型中,与对照组相比,0.5 mg/kg RAPA、0.75 mg/kg RAPA、1.0 mg/kg RAPA处理组肝组织重量均显著降低(P<0.05)。体外实验表明,与对照组相比,1μmol/L RAPA、3μmol/L RAPA、5μmol/L RAPA给药组细胞迁移能力明显降低(P<0.05);与对照组相比,3μmol/L RAPA、5μmol/L RAPA处理组侵袭到小室膜底侧细胞数明显减少(P<0.05);与对照组OD值比较,1μmol/L RAPA、3μmol/L RAPA、5μmol/L RAPA处理组细胞增殖活性显著降低(P<0.05);与对照组相比,3μmol/L RAPA、5μmol/L RAPA、10μmol/L RAPA干预的小鼠黑色素瘤细胞凋亡数显著增加(P<0.05)。结论:雷帕霉素能够抑制黑色素瘤的肝转移,该效应可能与黑色素瘤细胞迁移和侵袭能力降低有关。

Objective:To interrogate the potential of mTOR inhibitor rapamycin in intervention for melanoma metastasis.Methods:Liver metastasis models was established through intrasplenic injection of B16-F10 cells.Rapamycin was tested for the inhibitory effect on the development of metastatic lesions of melanoma,followed by pathological examination.MTT,migration,Transwell and flow cytometry assays were performed to evaluate the proliferation,migration,invasion and apoptosis of melanoma cells in the presence or absence of therapeutic agents.Results:In the liver metastasis model,compared with the control,the liver weight of the treatment groups treated with 0.5 mg/kg RAPA,0.75 mg/kg RAPA and 1.0 mg/kg RAPA were significantly decreased(P<0.05).Consistent with in vivo experiment,vitro experiments showed that compared with the control group,the number of migrating cells in the 1μmol/L RAPA group,3μmol/L RAPA group and 5μmol/L RAPA group were significantly decreased(P<0.05).Compared with the control group,the numbers of invading cells in the 3μmol/L RAPA group and 5μmol/L RAPA group were significantly decreased(P<0.05).Compared with the control group,the cell proliferation rate in the 1μmol/L RAPA group,3μmol/L RAPA group and 5μmol/L RAPA group were significantly decreased(P<0.05).Compared with the control group,the apoptotic rate of cells in the 3μmol/L RAPA group and 5μmol/L RAPA group and 10μmol/L RAPA group were significantly increased(P<0.05).Conclusion:Rapamycin inhibits liver metastasis of melanoma,which may be attributable to the suppressed capacity of migration and invasion of tumor cells.