目的 研究白花丹素(plumbagin,PL)对Lewis肺癌的抑制作用。方法 CCK8检测细胞的增殖;流式细胞仪检测细胞的凋亡率;Western blot检测细胞内Bcl-2和血管内皮生长因子(VEGF)蛋白的表达水平;建立Lewis肺癌小鼠模型;分别使用生理盐水、白花丹素(分小、中、大3个剂量组)及环磷酰胺(CTX)作用于动物模型;观察各组肿瘤体积大小、瘤重、抑瘤率及测肿瘤组织VEGF表达情况。结果 CCK8结果显示,白花丹素对Lewis肺癌细胞增殖有明显的抑制作用,且呈浓度依赖性(r=0.953,P<0.01);白花丹素显著增加Lewis肺癌细胞凋亡率(P<0.05);白花丹素可使Lewis肺癌细胞中Bcl-2和VEGF蛋白的表达较对照组显著下调(P<0.05);白花丹素小、中、大剂量组及CTX组的肿瘤体积、瘤重及肿瘤组织中VEGF蛋白表达水平均较对照组显著降低(P<0.05)。结论 白花丹素在体内外均能抑制Lewis肺癌细胞的增殖,其机制可能与下调Bcl-2、VEGF蛋白的表达和诱导Lewis肺癌细胞凋亡有关。
Abstract
OBJECITVE To study the inhibitive effect of plumbagin on Lewis lung cancer. METHODS Cell proliferation was determined by CCK8 assay. Apoptosis was determined by flow cytometry. The expression of Bcl-2 and VEGF protein was studied by Western blot assay. The model of C57BL/6 mice bearing Lewis lung cancer was established by subcutaneous seeding of Lewis lung cancer cells, and randomly divided into 5 groups (n=6). Tumor-bearing mice were injected with normal saline, plumbagin(low, medium, high dose) or cyclophosphamide (CTX) in each group. The tumor volume was measured. All mice were sacrificed on Day 22nd under aseptic condition for the tumor collection. The transplanted tumors were weighed for calculation of the tumor inhibition rate; Immunohistochemical method was applied to assessing the VEGF expression in tumor tissue. RESULTS CCK-8 assay showed that plumbagin had an obvious inhibition on Lewis lung carcinoma cells line in a dose-dependent manner(r=0.953, P<0.05). Plumbagin significantly increased cell apoptosis rate(P<0.05). The protein levels of Bcl-2 and VEGF were significantly reduced by plumbagin (0, 2.5, 5, 10 μmol·L-1) treatment(P<0.05). In plumbagin(low, medium, high dose) groups and CTX group, the tumour volume, tumour weight and the expression of VEGF were significantly less than those in the control group (P<0.05). CONCLUSION The plumbagin effectively inhibits Lewis lung carcinoma cells proliferation and tumor growth of Lewis lung carcinoma cells in mice. The mechanism involved is down-regulating the expression of Bcl-2 ,VEGF and inducing cell apoptosis.
关键词
白花丹素 /
Lewis 肺癌细胞 /
凋亡 /
肺癌移植瘤 /
血管内皮生长因子 /
Bcl-2
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Key words
plumbagin /
Lewis lung carcinoma cells /
apoptosis /
transplanted tumor of lung cancer /
vascular endothelial growth factor /
Bcl-2
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中图分类号：
R734.2
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脚注
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基金
江西省卫生和计划生育委员会中医药科研基金(2011A146)
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