CAJ | 학술논문

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-△CT值为1.2928±0.1509)明显高于M14细胞系(2-△CT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.
목적 탐토miRNA-21재인흑소류세포계A375、M14중적표체급대기세포활성적영향.방법 채용형광정량PCR법,이U6기인위내삼,분별검측A375、M14세포중miRNA-21적상대함량.이용LipofectamineTM2000지질체장삼충농도적miRNA-21억제제분별전염저량충세포계,용CCK-8검측전염전후실험조、대조조적세포활성.용SPSS13.0통계연건분석세포활성적변화정황.결과 형광정량PCR법검측제시,miRNA-21재A375세포계중적표체(2-△CT치위1.2928±0.1509)명현고우M14세포계(2-△CT치위0.1894±0.1803).miRNA-21억제제농도위90 nmol/L시,A375、M14세포활성수도겁현저억제,세포활성(R치)분별위0.7362±0.1662、0.7295±0.1478;당농도위180 nmol/L、270 nmol/L시,A375세포활성현저하강,R치분별위0.7248±0.3204、0.6767±0.2998,이M14세포활성무현저변화.결론 miRNA-21재A375、M14량충인흑소류세포계중균유불동정도적표체.miRNA-21재A375、M14세포계중가이촉진세포증식,가능하조료모사종류억제인자이기도암기인양작용.
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-deltaCT 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.