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目的:观察木鳖子醇提物(ethanol extract from cochinchina momordica seed,CMSEE)对小鼠黑素瘤细胞B16及其荷瘤小鼠移植瘤侵袭转移的影响,初步探讨其作用机制。方法:通过划痕实验和体外迁移实验检测不同质量浓度CMSEE(20、25、30μg/mL)作用24和48h后,B16细胞侵袭和转移能力的改变;分别用RT-PCR和Western印迹法检测不同质量浓度CMSEE(20、25、30μg/mL)作用24h以及25μg/mL CMSEE作用不同时间(0、6、12、24、36、48h)对B16细胞中基质金属蛋白酶(ma-trix metalloproteinase,MMP)-2、MMP-9mRNA及其蛋白以及基质金属蛋白酶组织抑制因子(tissue inhibitor of metalloproteinase,TIMP)-1、TIMP-2蛋白表达的影响。以B16细胞建立小鼠荷瘤模型,用10mg/kg CMSEE灌胃治疗荷瘤小鼠,HE染色观察各组小鼠的肿瘤、肝、脾组织的病理变化。结果:CMSEE(20~30μg/mL)可明显抑制B16细胞的侵袭和转移能力,可使B16细胞划痕的直径明显变宽(P<0.05),穿过Transwell小室膜的细胞数显著减少(P<0.05),并呈浓度依赖性;CMSEE(20~30μg/mL)可明显下调B16细胞中MMP-2、MMP-9mRNA及蛋白的表达(P<0.05),并有浓度依赖性;而CMSEE对B16细胞中TIMP-1和TIMP-2蛋白的表达无明显影响(P>0.05)。CMSEE可有效抑制小鼠体内黑素瘤的侵袭和转移。结论:CMSEE在体内外均能明显抑制小鼠B16细胞的侵袭和转移能力,其作用机制可能与其抑制MMP-2和MMP-9的表达有关。
OBJECTIVE: To investigate the effect of CMSEE on the invasion and metastasis of mouse melanoma B16 cells and its tumor-bearing mice xenografts and to explore its possible mechanism. Methods: The invasion and metastasis of B16 cells were detected by scratch test and in vitro migration assay at different concentrations of CMSEE (20, 25, 30 μg / mL) for 24 and 48 hours respectively. The differences were detected by RT-PCR and Western blot Effects of CMSEE (20,25,30μg / mL) for 24 hours and 25μg / mL CMSEE on the expression of matrine metalloproteinase (MMP) in B16 cells at different times (0,6,12,24,36,48h) -2, MMP-9mRNA and their proteins and the expression of tissue inhibitor of metalloproteinase (TIMP) -1 and TIMP-2. The tumor-bearing mice model was established by B16 cells and the tumor-bearing mice were treated with 10mg / kg CMSEE. The pathological changes of tumor, liver and spleen were observed by HE staining. Results: CMSEE (20 ~ 30μg / mL) significantly inhibited the invasion and metastasis of B16 cells, and significantly broadened the diameter of B16 cells (P <0.05) <0.05), and in a concentration-dependent manner. CMSEE (20-30μg / mL) could significantly down-regulate the expression of MMP-2, MMP-9mRNA and protein in B16cells (P <0.05) The expression of TIMP-1 and TIMP-2 in B16 cells had no significant effect (P> 0.05). CMSEE can effectively inhibit the invasion and metastasis of melanoma in mice. CONCLUSION: CMSEE can significantly inhibit the invasion and metastasis of B16 cells in vitro and in vivo, which may be related to the inhibition of the expression of MMP-2 and MMP-9.