中华皮肤科杂志 ›› 2009, Vol. 42 ›› Issue (3): 186-188.

• 论著 • 上一篇    下一篇

miRNA-21在人黑素瘤细胞系A375和M14中的表达及功能初探

王焱 孙建方 方方 熊慧 韩峻松 张国成   

  1. 中国医科院皮研所 南京医科院皮研所 南京医科院皮研所 国家人类基因组南方研究中心 生物芯片上海国家工程研究中心 南京医科院皮研所
  • 收稿日期:2008-04-02 修回日期:2008-04-24 出版日期:2009-03-15 发布日期:2009-03-15
  • 通讯作者: 王焱 E-mail:wang1890@yahoo.com.cn

A preliminary study on the expression and function of microRNA-21 in two human malignant melanoma cell lines, A375 and M14

  • Received:2008-04-02 Revised:2008-04-24 Online:2009-03-15 Published:2009-03-15

PDF (PC)

204

赞

摘要：

目的 探讨miRNA-21在人黑素瘤细胞系A375、M14中的表达及对其细胞活性的影响。 方法 采用荧光定量PCR法，以U6基因为内参，分别检测A375、M14细胞中miRNA-21的相对含量。利用LipofectamineTM2000脂质体将三种浓度的miRNA-21抑制剂分别转染这两种细胞系，用CCK-8检测转染前后实验组、对照组的细胞活性。用SPSS13.0统计软件分析细胞活性的变化情况。结果 荧光定量PCR法检测提示，miRNA-21在A375细胞系中的表达（2－ΔCT值为1.2928 ± 0.1509）明显高于M14细胞系（2－ΔCT值为0.1894 ± 0.1803）。miRNA-21抑制剂浓度为90 nmol/L时，A375、M14细胞活性受到极显著抑制，细胞活性（R值）分别为0.7362 ± 0.1662、0.7295 ± 0.1478；当浓度为180 nmol/L、270 nmol/L时，A375细胞活性显著下降，R值分别为0.7248 ± 0.3204、0.6767 ± 0.2998，而M14细胞活性无显著变化。结论 miRNA-21在A375、M14两种人黑素瘤细胞系中均有不同程度的表达。miRNA-21在A375、M14细胞系中可以促进细胞增殖，可能下调了某些肿瘤抑制因子而起到癌基因样作用。

关键词: 微小RNA;黑素瘤;细胞转染;抑制剂

Abstract:

Objective To investigate the expression of microRNA-21 （miRNA-21） in two human malignant melanoma cell lines, A375 and M14, and its effect on the viability of these cells. Methods The expression of human miRNA-21 was assessed by quantitative fluorescent PCR in A375 and M14 cells. Then, three concentrations （90, 180, 270 nmol/L） of miRNA-21 inhibitor and were transfected both cells and a negative control were transfected a mixture of LipofectamineTM 2000 reagent, respectively. After another 3-day culture, the proliferation of cells was detected by cell counting kit-8, and R value was calculated to denote the relative activity of cells. Statistical analysis was carried out by SPSS13.0. Results The expression of miRNA-21 was higher on A375 cells than that on M14 cells with the average value of 2-delta CT being 1.2928 ± 0.1509 vs 0.1894 ± 0.1803. With miRNA-21 inhibitor at the concentration of 90, 180, 270 nmol/L, the activity of A375 cells was significantly lowered in comparison with that in the control group, with the R value being 0.7362 ± 0.1662, 0.7248 ± 0.3204 and 0.6767 ± 0.2998 respectively （all P < 0.01）. However, in the case of M14 cells, cell activity was only suppressed by miRNA-21 inhibitor at 90 nmol/L with the R value being 0.7295 ± 0.1478, and no significant inhibition was observed with the inhibitor at 180 or 270 nmol/L （both P > 0.05）. Conclusions miRNA-21 is expressed on human melanoma cell lines, A375 and M14, at different levels, with a promoting effect on the proliferation of both cells. Moreover, miRNA-21 may act as an oncogene-like gene via down-regulating the expression of some tumor-inhibiting factors.

Key words: microRNA;Melanoma;Cell transfection;Inhibitor