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苦参碱在体外抑制人恶性黑色素瘤细胞系A375的侵袭和转移。

Matrine inhibits invasiveness and metastasis of human malignant melanoma cell line A375 in vitro.

作者信息

Liu Xiao-Yan, Fang Hong, Yang Zheng-Gang, Wang Xiao-Yong, Ruan Li-Ming, Fang De-Ren, Ding Ying-Guo, Wang Yi-Na, Zhang Yu, Jiang Xiao-Ling, Chen Hong-Chao

机构信息

Department of Dermatology, First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang, China.

出版信息

Int J Dermatol. 2008 May;47(5):448-56. doi: 10.1111/j.1365-4632.2008.03627.x.

Abstract

BACKGROUND

Matrine is a traditional Chinese medicine with significant inhibitory activity against malignant tumors. Its effects on the invasiveness and metastasis of malignant tumors have rarely been reported.

AIM

To investigate whether matrine can inhibit the metastasis-related activities of the human malignant melanoma cell line A375 in vitro.

METHODS

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and Annexin-V-fluorescein isothiocyanate/propidium iodide (Annexin-V-FITC/PI) affinity assay were used to examine the effects of matrine on the proliferation and apoptosis induction of A375 cells. The morphologic changes of A375 cells were observed by light and electron microscopy. Semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting were performed to evaluate the expression of heparanase mRNA and protein. The effect of matrine on the adhesion ability and invasiveness of treated A375 cells was tested by cell-Matrigel adhesion assay and Matrigel invasion assay, respectively.

RESULTS

Matrine showed significant inhibition of the proliferation of A375 cells in a dose- and time-dependent manner. It also induced apoptosis in a dose-dependent manner. Compared with the control group, the levels of heparanase mRNA and protein expression of A375 cells treated with different concentrations of matrine were decreased significantly, as were their adhesion ability and invasiveness.

CONCLUSIONS

These findings indicate that matrine inhibits the invasiveness and metastasis of A375 cells in vitro. The mechanisms may be linked to the inhibition of cellular proliferation, induction of apoptosis, and downregulation of heparanase mRNA and protein expression.

摘要

背景

苦参碱是一种对恶性肿瘤具有显著抑制活性的中药。其对恶性肿瘤侵袭和转移的影响鲜有报道。

目的

研究苦参碱在体外是否能抑制人恶性黑色素瘤细胞系A375的转移相关活性。

方法

采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)法和膜联蛋白-V-异硫氰酸荧光素/碘化丙啶(Annexin-V-FITC/PI)亲和法检测苦参碱对A375细胞增殖和凋亡诱导的影响。通过光学显微镜和电子显微镜观察A375细胞的形态变化。进行半定量逆转录-聚合酶链反应(RT-PCR)和蛋白质免疫印迹法以评估乙酰肝素酶mRNA和蛋白的表达。分别通过细胞-基质胶黏附试验和基质胶侵袭试验检测苦参碱对处理后的A375细胞黏附能力和侵袭能力的影响。

结果

苦参碱对A375细胞的增殖具有显著的剂量和时间依赖性抑制作用。它还以剂量依赖性方式诱导细胞凋亡。与对照组相比,不同浓度苦参碱处理的A375细胞中乙酰肝素酶mRNA和蛋白表达水平显著降低,其黏附能力和侵袭能力也显著降低。

结论

这些发现表明苦参碱在体外抑制A375细胞的侵袭和转移。其机制可能与抑制细胞增殖、诱导细胞凋亡以及下调乙酰肝素酶mRNA和蛋白表达有关。

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