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目的研究微小RNA-23a(miR-23a)对乳腺癌细胞增殖和侵袭的影响。方法将人乳腺癌MCF-7和MDA-MB-435S复苏培养后,接种至6孔板中,随机分组进行培养,共分为6组:RNAi-Ⅰ组、过表达-Ⅰ组、对照-Ⅰ组、RNAi-Ⅱ组、过表达-Ⅱ组、对照-Ⅱ组。构建MiR-23a重组干扰及过表达载体,用Lipofectamine 2000将构建成功的MiR-23a重组干扰及过表达载体分别转染至各组人乳腺癌MCF-7和MDA-MB-435S细胞系中。结果在培养72 h后的OD492值:过表达组的MCF-7细胞和MDA-MB-435S细胞可分别达到0.52±0.03和0.44±0.03,而RNAi组MCF-7细胞和MDA-MB-435S细胞仅为0.36±0.02和0.31±0.02,与对照-Ⅰ组和对照-Ⅱ组比较,RNAi-Ⅰ组和RNAi-Ⅱ组的差异均有统计学意义(均P<0.05)。过表达-Ⅰ组和过表达-Ⅱ组的细胞倍增时间分别为(26.61±5.23),(29.23±5.51)h,而RNAi-Ⅰ组和RNAi-Ⅱ组的细胞则分别为(42.24±3.47),(58.17±5.34)h。过表达-Ⅰ组和过表达-Ⅱ组的M期细胞比例分别为52.36%,51.59%;而RNAi-Ⅰ组和RNAi-Ⅱ组则分别为76.70%,74.17%。与对照-Ⅰ组比较,RNAi-Ⅰ组和过表达-Ⅰ组的细胞倍增时间及M期比例差异均有统计学意义(均P<0.05)。结论 MiR-23a的表达水平与乳腺癌细胞的增殖和侵袭呈现正相关性。
Objective To investigate the effect of microRNA-23a (miR-23a) on the proliferation and invasion of breast cancer cells. Methods Human breast cancer MCF-7 and MDA-MB-435S were resuscitated and cultured. The cells were inoculated into 6-well plates and randomly divided into 6 groups: RNAi-Ⅰ group, overexpression-Ⅰ group, Group, RNAi-Ⅱ group, overexpression-Ⅱ group, control-Ⅱ group. MiR-23a was constructed and transfected into human breast cancer MCF-7 and MDA-MB-435S cells by Lipofectamine 2000. The recombinant MiR-23a was constructed and transfected into human breast cancer cell lines MCF-7 and MDA-MB-435S respectively. Results The OD492 value of 72 h after culture showed that the expression of MCF-7 cells and MDA-MB-435S cells in overexpression group reached 0.52 ± 0.03 and 0.44 ± 0.03, respectively, while in RNAi group MCF-7 and MDA-MB-435S cells Only 0.36 ± 0.02 and 0.31 ± 0.02. There were significant differences between RNAi-Ⅰ group and RNAi-Ⅱ group (all P <0.05) compared with the control -Ⅰgroup and the control -Ⅱgroup. The cell doubling time was (26.61 ± 5.23) and (29.23 ± 5.51) h in overexpression-Ⅰ group and overexpression-Ⅱ group, while it was (42.24 ± 3.47) times in RNAi-Ⅰ group and RNAi-Ⅱ group , (58.17 ± 5.34) h. The proportion of M phase cells in over-expression groupⅠand overexpression-Ⅱ group were 52.36% and 51.59% respectively, while the RNAi-Ⅰ group and RNAi-Ⅱ group were 76.70% and 74.17% respectively. Compared with the control-Ⅰ group, the differences of the doubling time and the M-phase ratio between RNAi-Ⅰ group and overexpression-Ⅰ group were statistically significant (all P <0.05). Conclusion The expression of MiR-23a is positively correlated with the proliferation and invasion of breast cancer cells.