微小RNA-133a-3p对乳腺癌细胞的影响及机制研究

Effect and mechanism of microRNA-133A-3p on breast cancer cells

目的探究微小RNA-133a-3p(miR-133a-3p)通过靶向调控CAND蛋白1(CAND1)对乳腺癌细胞侵袭及凋亡的影响及机制。方法MCF-7细胞分为过表达组(转染mimics-miR-133a-3p)、NC组(转染mimics对照)、共转染组(共同转染mimics-miR-133a-3p和pcDNA-CAND1)、对照组(仅加入等量转染试剂)。以流式细胞仪检测细胞凋亡情况,以Transwell实验检测细胞侵袭能力,以实时荧光定量聚合酶链反应检测miR-133a-3p和CAND1的表达。结果转染后,对照组、NC组、过表达组及共转染组miR-133a-3p表达水平分别为0.50±0.08、0.51±0.09、1.06±0.10和1.05±0.15,CAND1 mRNA的表达分别为0.91±0.09、0.91±0.07、0.80±0.10和1.21±0.10。上述指标,共转染组与对照组、NC组比较,在统计学上差异均有统计学意义(均P<0.05),过表达组与对照组、NC组比较,在统计学上差异均有统计学意义(均P<0.05)。对照组、NC组、过表达组及共转染组中细胞凋亡率分别为(7.88±1.62)%、(8.87±2.01)%、(53.41±5.46)%和(29.54±3.78)%;对照组、NC组、过表达组及共转染组中细胞侵袭数分别为(161.02±10.31)、(155.87±12.30)、(85.21±9.11)和(118.37±10.84)个。上述指标,转染组与过表达组、对照组、NC组比较,在统计学上差异均有统计学意义(均P<0.05),过表达组与对照组、NC组比较,在统计学上差异均有统计学意义(均P<0.05)。结论在人乳腺癌细胞MCF-7中过表达miR-133a-3p可抑制CAND1,促进MCF-7细胞凋亡和侵袭。

Objective To explore the effect and mechanism of microRNA-133a-3p(miR-133a-3p)on invasion and apoptosis of breast cancer cells through targeted regulation of cullin-associated NEDD8-dissociated 1(CAND1).Methods MCF-7 cells were divided into overexpression group(mimics miR-133a-3p transfection),NC group(mimics control transfection),co-transfection group(mimics miR-133a-3p transfection with pcDNA-CAND1 co-transfection)and control group(only adding the same amount of transfection reagents).Flow cytometry was used to detect cell apoptosis,Transwell assay was used to detect cell invasion,and real-time fluorescence quantitative polymerase chain reaction was used to detect miR-133a-3p and CAND1 expression.Results After transfection,the expression levels of miR-133a-3p in control group,NC group,overexpression group and co-transfection group were 0.50±0.08,0.51±0.09,1.06±0.10 and 1.05±0.15,respectively;the expression of CAND1 mRNA were 0.91±0.09,0.91±0.07,0.80±0.10 and 1.21±0.10,respectively.There were statistically significant differences in the above indexes between the co-transfection group and the control group,the NC group(all P<0.05),and there were statistically significant differences between the overexpression group and the control group,the NC group(all P<0.05).The apoptosis rates in control group,NC group,overexpression group and co-transfection group were(7.88±1.62)%,(8.87±2.01)%,(53.41±5.46)%,(29.54±3.78)%,respectively.The number of invasive cells in control group,NC group,overexpression group and co-transfection group were 161.02±10.31,155.87±12.30,85.21±9.11 and 118.37±10.84,respectively.There were statistically significant differences in the above indexes between transfection group and overexpression group,control group and NC group(all P<0.05),and there were statistically significant differences between overexpression group and control group and NC group(all P<0.05).Conclusion Overexpression of miR-133a-3p in human breast cancer cells MCF-7 can inhibit CAND1 and promote apoptosis and invasion of MCF-7 cells.