Liang Ai-Ling, Zhang Ting-Ting, Zhou Ning, Wu Cui Yun, Lin Man-Hua, Liu Yong-Jun
Medical Molecular Diagnostics Key Laboratory of Guangdong, and Departments of Biochemistry and Molecular Biology and Clinical Biochemistry, Guangdong Medical University, Dongguan, Guangdong 523808, P.R. China.
Department of Clinical Laboratory, Huangshi City Central Hospital, Huangshi, Hubei 415000, P.R. China.
Oncol Rep. 2016 Apr;35(4):1950-8. doi: 10.3892/or.2016.4596. Epub 2016 Jan 26.
Breast cancer is a malignant tumor with the highest incidence among women. Breast cancer metastasis is the major cause of treatment failure and mortality among such patients. MicroRNAs (miRNAs) are a class of small molecular non-coding regulatory RNAs, which act as oncogenes or tumor suppressors in breast cancer. miRNA-10b has been found to exhibit a high expression level in advanced and metastatic breast cancer, and is closely related to breast cancer metastasis. An miRNA sponge is an mRNA with several repeated sequences of complete or incomplete complementarity to the natural miRNA in its 3' non-translating region. It acts as a sponge adsorbing miRNAs and ensures their separation from their targets and inhibits their function. The present study designed a sponge plasmid against miRNA-10b and transiently transfected it into high and low metastatic human breast cancer cell lines MDA-MB-231 and MCF-7, and analyzed the effects of the miRNA-10b sponge on the growth and proliferation, migration and invasion in these cell lines. qRT-PCR results found that the sponge plasmid effectively inhibited the expression of miRNA-10b, and upregulated the expression of the miRNA‑10b target protein HOXD-10. The results from the CCK-8 assay found that the miRNA-10b sponge inhibited the growth of breast cancer cell lines MDA-MB-231 and MCF-7. Results of the plate cloning experiments indicated that the miRNA-10b sponge suppressed the colony formation of the MDA-MB-231 and MCF-7 cells. The results of wound healing and Transwell assays showed that the miRNA-10b sponge inhibited the migration and invasion of the breast cancer cell lines MDA-MB-231 and MCF-7. Our results demonstrated that the miRNA-10b sponge effectively inhibited the growth and proliferation of breast cancer MDA-MB-231 and MCF-7 cells. In addition, it also restrained the migration and invasion of human highly metastatic breast cancer MDA-MB-231 cells.
乳腺癌是女性中发病率最高的恶性肿瘤。乳腺癌转移是这类患者治疗失败和死亡的主要原因。微小RNA(miRNA)是一类小分子非编码调节RNA,在乳腺癌中可作为癌基因或肿瘤抑制因子发挥作用。已发现miRNA - 10b在晚期和转移性乳腺癌中呈现高表达水平,且与乳腺癌转移密切相关。miRNA海绵是一种mRNA,其3'非翻译区具有与天然miRNA完全或不完全互补的多个重复序列。它起到海绵吸附miRNA的作用,确保它们与靶标分离并抑制其功能。本研究设计了针对miRNA - 10b的海绵质粒,并将其瞬时转染到高转移和低转移的人乳腺癌细胞系MDA - MB - 231和MCF - 7中,分析了miRNA - 10b海绵对这些细胞系生长增殖、迁移和侵袭的影响。qRT - PCR结果发现,海绵质粒有效抑制了miRNA - 10b的表达,并上调了miRNA - 10b靶蛋白HOXD - 10的表达。CCK - 8检测结果发现,miRNA - 10b海绵抑制了乳腺癌细胞系MDA - MB - 231和MCF - 7的生长。平板克隆实验结果表明,miRNA - 10b海绵抑制了MDA - MB - 231和MCF - 7细胞的集落形成。伤口愈合和Transwell检测结果显示,miRNA - 10b海绵抑制了乳腺癌细胞系MDA - MB - 231和MCF - 7的迁移和侵袭。我们的结果表明,miRNA - 10b海绵有效抑制了乳腺癌MDA - MB - 231和MCF - 7细胞的生长和增殖。此外,它还抑制了人高转移性乳腺癌MDA - MB - 231细胞的迁移和侵袭。