天津医药 ›› 2018, Vol. 46 ›› Issue (12): 1273-1276.doi: 10.11958/20181063

• 细胞与分子生物学 • 上一篇    下一篇

去甲斑蝥素抑制黑色素瘤M14细胞增殖和诱导细胞凋亡的研究

高杨,程毅,任明媛,岳育杨   

  1. 1石家庄,河北省儿童医院介入血管科(邮编050000);2石家庄,河北医科大学第四医院皮肤科
  • 收稿日期:2018-07-13 修回日期:2018-09-03 出版日期:2018-12-15 发布日期:2019-01-24
  • 通讯作者: 高杨 E-mail:gaoyang507@163.com

The inhibitory effects of norcantharidin on proliferation and apoptosis of melanoma M14 cells

GAO Yang, CHENG Yi, REN Ming-yuan, YUE Yu-yang   

  1. 1 Department of Interventional Radiology, Children's Hospital of Hebei Province, Shijiazhuang 050000, China; 2 Department of Dermatology, the Forth Hospital of Hebei Medical University
  • Received:2018-07-13 Revised:2018-09-03 Published:2018-12-15 Online:2019-01-24

摘要: Objective To investigate the effects of norcantharidin on the proliferation and apoptosis related proteins Bax and Bcl-2 of human melanoma M14 cells. Methods Melanoma M14 cell line cultured in vitro was treated with concentrations of 50, 100 and 500 μg / L norcantharidin respectively, and untreated cells were used as control. Cell proliferation was detected by CCK-8 method after treatment for 24 h and 48 h. Morphology of cells was observed under inverted microscope. DAPI nuclear staining was used to observe the morphological changes of apoptosis under fluorescence microscope. The expressions of Bax and Bcl-2 were detected by Western blot assay. Results CCK-8 results showed that concentrations of 100 and 500 μg/L norcantharidin inhibited the proliferation of melanoma M14 cells after treatment for 24 h and 48 h (P<0.05), and the higher the concentration, the more significant the inhibitory effect. The cells showed irregular contours, nucleus shrinkage, small volume, and poor cell adhesion of apoptosis changes under inverted microscope and fluorescence microscope. Western blot analysis showed that the relative expression of Bcl-2 decreased gradually with the increased concentration of norcantharidin (P<0.05), and the relative expression of Bax increased gradually (P<0.05).Conclusion Norcantharidin can inhibit the proliferation of melanoma M14 cells, and its mechanism may be related to the promotion of apoptosis.

关键词: 黑色素瘤, 实验性, 细胞凋亡, 抗肿瘤药, 植物, Bcl-2相关X蛋白质, 去甲斑蝥素, Bax

Abstract: Objective To investigate the effects of norcantharidin on the proliferation and apoptosis related proteins Bax and Bcl-2 of human melanoma M14 cells. Methods Melanoma M14 cell line cultured in vitro was treated with concentrations of 50, 100 and 500 μg / L norcantharidin respectively, and untreated cells were used as control. Cell proliferation was detected by CCK-8 method after treatment for 24 h and 48 h. Morphology of cells was observed under inverted microscope. DAPI nuclear staining was used to observe the morphological changes of apoptosis under fluorescence microscope. The expressions of Bax and Bcl-2 were detected by Western blot assay. Results CCK-8 results showed that concentrations of 100 and 500 μg/L norcantharidin inhibited the proliferation of melanoma M14 cells after treatment for 24 h and 48 h (P<0.05), and the higher the concentration, the more significant the inhibitory effect. The cells showed irregular contours, nucleus shrinkage, small volume, and poor cell adhesion of apoptosis changes under inverted microscope and fluorescence microscope. Western blot analysis showed that the relative expression of Bcl-2 decreased gradually with the increased concentration of norcantharidin (P<0.05), and the relative expression of Bax increased gradually (P<0.05).Conclusion Norcantharidin can inhibit the proliferation of melanoma M14 cells, and its mechanism may be related to the promotion of apoptosis.

Key words: melanoma, experimental, apoptosis, antineopl agents, phytogenic, Bcl-2-associated X protein, norcantharidin, Bax