Hao Chunfang, Wei Sen, Tong Zhongsheng, Li Shufen, Shi Yehui, Wang Xiaorui, Zhu Zhi-hua
Department of Breast Oncology, Tianjin Medical University Cancer Institute and Hospital, Key Laboratory of Breast Cancer Prevention and Therapy, Tianjin Medical University, Ministry of Education, Key Laboratory of Cancer Prevention and Therapy, Huanhuxi Road, Hexi District, Tianjin 300060, China.
Tumour Biol. 2012 Aug;33(4):1159-67. doi: 10.1007/s13277-012-0358-7. Epub 2012 Feb 29.
The purpose of this study was to investigate the effect of Raf kinase inhibitor protein (RKIP) on the growth, proliferation, invasion and metastasis of triple-negative breast cancer (TNBC) cells to provide experimental evidence for developing future therapies against human TNBC. The pcDNA3.1-RKIP eukaryotic expression vector was constructed and transfected into the TNBC cell line MDA-MB-231. The alterations of the biological characteristics of RKIP-transfected MDA-MB-231 cells were analyzed using the following approaches: a growth curve, a 3-(4,5-dimethylthiazol-2-Yl)-2,5-diphenyltetrazolium bromide (MTT) assay, bromodeoxyuridine (BrdU) staining and a cell migration assay. The effects of the RKIP gene on MMP-1 and MMP-2 expression were also examined. The pcDNA3.1 empty vector-transfected and mock-transfected MDA-MB-231 cells were used as control groups. Compared with the empty vector-transfected and mock-transfected cells, the cell growth of RKIP-transfected MDA-MB-231 cells was significantly reduced. The empty vector-transfected group was not significantly different compared with the mock-transfected MDA-MB-231 cells. The results of the MTT and BrdU assays demonstrated that the proliferation of pcDNA3.1-RKIP-transfected cells was significantly reduced compared to the control cells (P < 0.05). The result of the cell migration assay suggested that the cross-membrane migration rate of the pcDNA3.1-RKIP-transfected cells was significantly lower than that of the control MDA-MB-231 cells (P < 0.05). We also demonstrated that RKIP may inhibit MMP-1 and MMP-2 expression in MDA-MB-231 cells. The RKIP gene may play a role in inhibiting cellular proliferation. The RKIP gene may also have some inhibitory effects on the invasiveness and metastatic capability of human TNBC cells.
本研究旨在探讨Raf激酶抑制蛋白(RKIP)对三阴性乳腺癌(TNBC)细胞生长、增殖、侵袭和转移的影响,为开发未来针对人类TNBC的治疗方法提供实验依据。构建了pcDNA3.1-RKIP真核表达载体,并将其转染至TNBC细胞系MDA-MB-231中。采用以下方法分析RKIP转染的MDA-MB-231细胞生物学特性的改变:生长曲线、3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)法、溴脱氧尿苷(BrdU)染色和细胞迁移试验。还检测了RKIP基因对MMP-1和MMP-2表达的影响。将pcDNA3.1空载体转染和模拟转染的MDA-MB-231细胞用作对照组。与空载体转染和模拟转染的细胞相比,RKIP转染的MDA-MB-231细胞的生长明显受到抑制。空载体转染组与模拟转染的MDA-MB-231细胞相比无显著差异。MTT和BrdU试验结果表明,与对照细胞相比,pcDNA3.1-RKIP转染细胞的增殖明显降低(P<0.05)。细胞迁移试验结果表明,pcDNA3.1-RKIP转染细胞的跨膜迁移率明显低于对照MDA-MB-231细胞(P<0.05)。我们还证明RKIP可能抑制MDA-MB-231细胞中MMP-1和MMP-2的表达。RKIP基因可能在抑制细胞增殖中发挥作用。RKIP基因对人TNBC细胞的侵袭性和转移能力也可能有一定的抑制作用。