MicroRNA-203 suppresses cell proliferation and migration by targeting BIRC5 and LASP1 in human triple-negative breast cancer cells

Journal of Experimental & Clinical Cancer Research, Jun 2012

This study was performed to investigate the effect of microRNA-203 (miR-203) on cell proliferation and migration in triple-negative breast cancer (TNBC). Real-time PCR was performed to detect the expression of miR-203 in TNBC cell lines. miR-203 precursor and control microRNA (miRNA) were transfected into triple-negative breast cancer (TNBC) cell lines and the effects of miR-203 up-regulation on the proliferation and migration of cells were investigated. Meanwhile, the mRNA and protein levels of baculoviral IAP repeat-containing protein 5 (BIRC5) and Lim and SH3 domain protein 1 (LASP1) were measured. Luciferase assays were also performed to validate BIRC5 and LASP1 as miR-203 targets. Both miR-203 and BIRC5 siRNA signicantly inhibited cell proliferation in TNBC cells. Both miR-203 and LASP1 siRNA signicantly inhibited cell migration in TNBC cells, also. Moreover, up-regulated of BIRC5 and LASP1 was able to abrogate the effects induced by transfection with the miR-203 precursor. These data suggest that miR-203 may function as a tumor suppressor in TNBC cells. Thus, miR-203 could be a potential therapeutic target for this disease.

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MicroRNA-203 suppresses cell proliferation and migration by targeting BIRC5 and LASP1 in human triple-negative breast cancer cells

Chen Wang Xiangqian Zheng Chunyan Shen Yurong Shi 0 0 Tianjin Cancer Institute , Huanhuxi Ave, Tianjin 300060 , China Background: This study was performed to investigate the effect of microRNA-203 (miR-203) on cell proliferation and migration in triple-negative breast cancer (TNBC). Methods: Real-time PCR was performed to detect the expression of miR-203 in TNBC cell lines. miR-203 precursor and control microRNA (miRNA) were transfected into triple-negative breast cancer (TNBC) cell lines and the effects of miR-203 up-regulation on the proliferation and migration of cells were investigated. Meanwhile, the mRNA and protein levels of baculoviral IAP repeat-containing protein 5 (BIRC5) and Lim and SH3 domain protein 1 (LASP1) were measured. Luciferase assays were also performed to validate BIRC5 and LASP1 as miR-203 targets. Results: Both miR-203 and BIRC5 siRNA signicantly inhibited cell proliferation in TNBC cells. Both miR-203 and LASP1 siRNA signicantly inhibited cell migration in TNBC cells, also. Moreover, up-regulated of BIRC5 and LASP1 was able to abrogate the effects induced by transfection with the miR-203 precursor. Conclusions: These data suggest that miR-203 may function as a tumor suppressor in TNBC cells. Thus, miR-203 could be a potential therapeutic target for this disease. - Background Breast cancer is the most frequently diagnosed cancer and the leading cause of cancer death in women worldwide, accounting for 23% (1.38 million) of all new cancer cases and 14% (458,400) of all cancer deaths in 2008. Approximately half of all breast cancer cases and 60% of breast cancer-related deaths are estimated to occur in developing countries [1]. The large number of etiological factors and the complexity of breast cancer present challenge for prevention and treatment. Triple-negative breast cancer (TNBC) is defined histologically as invasive carcinoma of the breast that lacks staining for estrogen receptor (ER), progesterone receptor (PgR), and the human epidermal growth factor receptor-2 (HER2). TNBC is associated with high proliferative rates, early recurrence, and poor survival rates. Much effort has been spent on the study of the biological behavior of TNBC cells to develop effective treatment strategies. MicroRNAs (miRNAs) are small, non-coding RNAs of 1925 nucleotides in length that are endogenously expressed in mammalian cells. miRNAs regulate gene expression post-transcriptionally, by pairing with complementary nucleotide sequences in the 3-UTRs of specific target mRNAs [2,3]. This recently identified type of gene regulators is involved in modulating multiple cellular pathways, including cell proliferation, differentiation, and migration. Thus, miRNAs may function as oncogenic miRNAs or tumor suppressors [4-6]. Over 50% of miRNA genes are located in cancer-associated genomic regions [7]. The deletion or epigenetic silencing of a miRNA that normally represses the expression of one or more oncogenes might lead to carcinogenesis, tumor growth and invasion, as has been demonstrated for miR200, miR-122 and miR-203 [8-10]. miR-203 is significantly down regulated in several cancers, including hepatocellular carcinoma [11], colon cancer [12], prostate cancer [13], and laryngeal cancer [14]. Because the down-regulated of miR-203 is common to a number of cancers, it has been hypothesized that miR203 may play an important role in tumorigenesis and tumor development. However, the function of miR-203 in breast cancer remains unclear, especially in TNBC. In this paper, we showed that miR-203 was downregulated in TNBC cell lines and that the ectopic overexpression of miR-203 blocked tumor cell proliferation and migration in vitro. Furthermore, BIRC5 and LASP1 were identified as two direct functional targets of miR203 in TNBC cells. These data suggest that the reduced expression of miR-203 facilitates the development and metastasis of TNBC. Materials and methods Cell culture and treatment Human triple-negative breast cancer cell lines (MDAMB-468 and MDA-MB-231) and normal breast cell line MCF-10A, were purchased from the American Type Culture Collection. MDA-MB-468 and MDA-MB-231 cells were maintained in DMEM (Gibco) supplemented with 10% FBS and 100 U/ml penicillin and 100 g/ml streptomycin. MCF-10A cells were maintained in DMEM/F-12 supplemented with 10% FBS, insulin (10 g /ml), hydrocortisone (500 ng/ml) and EGF (20 ng/ml). The cells were collected using 0.05% trypsin EDTA following the specified incubation period. Precursor miRNA/siRNA/plasmid transfection Cells were seeded in 6-well plates at a concentration of 1 105 and cultured in medium without antibiotics for approximately 24 h before transfection. Cells were transiently transfected with miR-203 precursor (Applied Biosystems) or negative control miRNA, BIRC5 siRNA (Sigma), LASP1 siRNA (Sigma) or control siRNA at a final concentration of 200nM. PcDNA-BIRC5 or pcDNA-LASP1 plasmid was also transfected into MDAMB-231 cells using Lipofectamine 20 (...truncated)


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Chen Wang, Xiangqian Zheng, Chunyan Shen, Yurong Shi. MicroRNA-203 suppresses cell proliferation and migration by targeting BIRC5 and LASP1 in human triple-negative breast cancer cells, Journal of Experimental & Clinical Cancer Research, 2012, pp. 58, Volume 31, Issue 1, DOI: 10.1186/1756-9966-31-58