苦参碱对K562细胞Cgi-100基因表达和细胞增殖的影响

Effects of Matrine on Cgi-100 Gene Expression and Proliferation in K562 Cells

本研究了解功能未知的cgi-100基因在苦参碱作用人白血病K562细胞中的表达情况,并探讨其对K562细胞生长的影响。应用RT-PCR检测苦参碱作用前后K562细胞中cgi-100基因的表达情况;构建pIRES2-EGFP/cgi-100真核表达质粒,用脂质体法转染至K562细胞中;RT-PCR检测在重组细胞中目的基因cgi-100的表达状况,并采用台盼蓝染色法观察细胞生长趋势,电镜观察细胞形态学改变,流式细胞术检测细胞周期的变化,观察cgi-100基因过表达对K562细胞增殖的影响。结果表明,在苦参碱作用K562细胞后cgi-100基因表达下调;重组K562细胞中cgi-100基因过表达,且常染色质增加,异染色质减少,细胞增殖旺盛,S期细胞增多,增殖速度高于对照组。结论:不同浓度不同作用时间下苦参碱能使K562细胞中cgi-100基因表达下降;cgi-100基因过表达能使K562细胞增殖加速、细胞幼稚程度增加。

This study was aimed to explore the expression level of the unknown cgi-100 gene in human leukemia K562 cells treated with matrine, and to investigate effect of cgi-100 on proliferation of K562 cells. The expression level of cgi-100 was detected by RT-PCR in K562 cells before and after being treated with matrine; pIRES2-EGFP/cgi-100 eukaryotic expression vector was constructed by DNA recombinant technique and was introduced into K562 cells by liposome-mediated DNA transfection. The cgi-100 gene expression level, growth-curve, and cell cycle of the modified K562-cgi-100 cells were detected by RT-PCR, Trypan blue staining and FCM. Morphological changes were observed under the optical and electron microscopes. The results indicated that the expression level of cgi-100 decreased in K562 cells treated with matrine. Heterochromatin decreased, euchromatin and the proportion of S phase in K562-cgi-100 cells increased, and cell proliferation enhanced. It is concluded that the expression of cgi-100 mRNA decreased in a dose- and time-dependent manner in the K562 cells treated with matrine and over-expression of cgi-100 elevates the proliferation and the immaturity level of K562- cgi-100 cells.