摘要
目的:探讨微小RNA(microRNA-214,miR-214)在人结肠癌HCT116细胞中的表达,以及其对HCT116细胞增殖及凋亡等生物学功能的影响。方法:采用实时荧光定量-PCR检测HCT116细胞中miR-214和PIM1 mRNA的表达情况,并分析二者的相关性。脂质体转染法将miR-214模拟物(mimics)转入HCT116细胞,用实时荧光定量-PCR法检测miR-214和PIM1 mRNA的表达情况;MTT法和平板克隆形成实验检测细胞增殖能力的改变情况;FCM法检测对细胞凋亡及细胞周期的影响。结果:MiR-214在结肠癌细胞株中低表达。结肠癌细胞中miR-214和PIM1 mRNA的表达呈负相关(r=-0.943,P=0.016)。MiR-214 mimics组HCT116细胞中miR-214表达量较对照组明显上调(P<0.01);miR-214高表达可以显著抑制PIM1 mRNA的表达水平。MiR-214 mimics组细胞的克隆数较对照组明显减少(P=0.026 9),细胞生长活力明显受到抑制(P<0.000 1),同时细胞凋亡率明显增加(P=0.010 4),G1期所占细胞数减少,而S期所占细胞数增加。结论:MiR-214可以抑制结肠癌细胞HCT116的增殖并促进其凋亡,miR-214作为一种抑癌因子,可能通过抑制PIM1在结肠癌中发挥抑癌作用。
Objective： To investigate the expression of miR-214 （microRNA-214） in different colon cancer cell lines and its effect on the proliferation and apoptosis of HCT116 cells. Methods： The expressions of miR-214 and PIM1 mRNA in different colon cancer cell lines were detected by real-time fluorogenic quantitative-PCR, and the relationship between miR-214 and PIM1 mRNA expression was evaluated. The miR-214 mimics was transfected into HCT116 cells by LipofectAMINE 2000, then the expressions of miR-214 and PIM1 mRNA levels were detected by real-time fluorogenic quantitative-PCR. The change of proliferation ablility of HCT116 cells was detected by MTT method and colony-formation assay. The flow cytometry was used to examine the changes of apoptosis and cell cycle distribution. Results： The expression of miR-214 was down-regulated in HCT116 cells. The expression of miR-214 was negatively correlated with the expression of PIM1 mRNA in colon cancer cells （r = –0.943, P = 0.016）. After transfection with miR-214 mimics, the expression level of miR-214 was up-regulated as compared with no transfection. The high expression level of miR-214 increased in miR-214 mimics group compared with that in the negative control group （P 〈 0.01）. The high expression level of miR-214 could significantly inhibit the expression of PIM1 mRNA. After transfection with miR-214 mimics, the number of colonies was decreased （P = 0.026 9）, the cell growth was inhibited （P 〈 0.000 1）, the apoptosis rate was increased （P = 0.010 4）, and the proportion of G1-stage cells was decreased as well as the proportion of S-stage cells was increased. Conclusion： MiR-214 can inhibit the proliferation and promote the apoptosis of colon cancer HCT116 cells. MiR-214 may act as a tumor suppressor in colorectal cancer by inhibiting PIM1.
出处
《肿瘤》
CAS
CSCD
北大核心
2013年第11期966-972,共7页
Tumor
基金
国家自然科学基金资助项目(编号:81072034)
河北省科技计划资助项目(编号:12396105D)
