下调miR-148a-3p表达对乳腺癌细胞增殖和迁移侵袭影响

Effects of down-regulation of miR-148a-3p on proliferation,migration and invasion of breast carcinoma cells

目的复发和转移是阻碍乳腺癌治疗效果的主要原因,研究复发转移机制是提高疗效的关键。本研究探讨miR-148a-3p在乳腺癌细胞系中表达,并研究抑制miR-148a-3p表达对人乳腺癌细胞增殖、迁移和侵袭能力影响。方法实时荧光定量PCR检测乳腺癌细胞系中miR-148a-3p表达,转染miR-148a-3p抑制物通过实时荧光定量PCR检测其抑制效率。细胞计数试剂盒CCK-8法和Transwell实验分别检测miR-148a-3p对乳腺癌细胞MCF-7和MDA-MB-231增殖、迁移和侵袭影响。调取并分析TCGA数据库中miR-148a-3p在乳腺癌肿瘤组织中表达。结果实时荧光定量PCR检测结果发现,miR-148a-3p RNA相对表达量在MCF-7乳腺癌细胞为1.430±0.093,MDA-MB-231乳腺癌细胞为1.370±0.081,高于正常乳腺癌上皮细胞MCF10A(1.013±0.062),t值分别为14.233、16.509,均P<0.001。CCK-8实验结果表明,抑制miR-148a-3p表达后,MCF-7和MDA-MB-231乳腺癌细胞系增殖能力下降。Transwell迁移实验结果显示,乳腺癌细胞系迁移细胞数目miR-148a-3p inhibitor组MCF-7为61.010±7.941,少于miR-NC组102.327±7.092,差异有统计学意义,t=6.725,P<0.001;乳腺癌细胞系迁移细胞数目miR-148a-3p inhibitor组MDA-MB-231为64.332±6.110,少于miR-NC组109.332±5.687,t=9.338,P<0.001;Transwell侵袭实验结果显示,乳腺癌细胞系迁移细胞数目miR-148a-3p inhibitor组MCF-7为42.331±4.042,少于miR-NC组细胞侵袭数目81.130±14.112,t=4.564,P<0.001;MDA-MB-231乳腺癌细胞系侵袭细胞数目为41.328±2.520,少于miR-NC组细胞侵袭数目82.671±6.657,t=10.06,P<0.001。结论 miR-148a-3p在乳腺癌细胞中高表达,并影响乳腺癌细胞的增殖、侵袭和迁移能力。

OBJECTIVE Recurrence and metastasis are the major factors affecting the curative effect of breast carcinoma.Investigating the mechanism of recurrence and metastasis is the key to improve the efficacy.In this study,we aimed to explore the expression and biological function of MicroRNA 148 a-3 p(miR-148 a-3 p)in breast carcinoma.METHODS Quantitative Real-time PCR(qRT-PCR)was performed to examine the expression of miR-148 a-3 p in breast carcinoma cell lines.The inhibition efficiency after transfection with miR-148 a-3 p inhibitor was examined by qRT-PCR.CCK-8 assay and transwell assay were used to detect the effect of miR-148 a-3 p on proliferation,migration and invasion of breast carcinoma cells.RESULTS The qRT-PCR results showed that the expression of miR-148 a-3 p in breast carcinoma cell lines MCF-7 was 1.430±0.093,and in breast carcinoma cell lines MDA-MB-231 was 1.370±0.081,which were both higher than that in breast normal epithelial cell line MCF10 A(1.013±0.062),t=14.233,16.509,and both P< 0.001.CCK-8 assay revealed that the decreased expression of miR-148 a-3 p reduced the proliferative ability of MCF-7 and MDA-MB-231.Transwell migration test showed that the number of migrated cells in MCF-7 breast cancer cell lines in the miR-148 a-3 p inhibitor group was 61.010±7.941,which was significantly lower than the number of migrated cells in the miR-NC group(102.327±7.092),t=6.725,P<0.001.The number of migrated cells in MDA-MB-231 breast cancer cell lines in the miR-148 a-3 p inhibitor group was 64.332±6.110,which was lower than that of the miR-NC group(109.331±5.687),t=9.338,P<0.001.Transwell invasion test showed that the number of migrated cells in MCF-7 breast cancer cell lines in the miR-148 a-3 p inhibitor group was 42.331±4.042,which was significantly lower than that of the miR-NC group(81.130±14.112),t=4.564,P<0.001.The number of invasion cells in MDA-MB-231 breast cancer cell line was 41.328±2.520,which was lower than that of the miR-NC group(82.671±6.657),t=10.063,P<0.001.CONCLUSION MiR-148 a-3 p is highly expressed in breast carcinoma cells and tumor tissues which affect the breast carcinoma cells’ proliferation,migration and invasion ability.