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Differential expression of miR-200c in breast cancer stem cells.

ABSTRACT: Breast cancer has become the most popular malignant tumors in females. Breast cancer stem cells (BCSCs) play an important role in metastasis and recurrence. Previous study showed close correlation between miR-200c and behavioral regulation of breast cancer stem cells. This study aimed to investigate differential expression of miR-200c in breast cancer stem cells, and its role in regulation tumor formation property. DNA microchip assay and qRT-PCR analyzed differential expression of microRNA between BCSCs and non-tumor formation NTG cells. Bioinformatics and luciferase reporter gene assay determined targeted site of miR-200c. MiR mimic and inhibitor were transfected to change miR-200c expression level, followed by Western Blot for detecting BMI1 gene expression. Cell clonal assay determined cell proliferation, whilst tumor xenograft was performed to analyze tumor formation potency. MiR-200c was down-regulated in BCSCs compared to NTG cells (P<0.05). Bioinformatics and luciferase reporter gene assay attributed BMI1 gene as targeting site of miR-200c. Transfection of miR-200c and inhibitor altered miR-200c expression. Results showed that miR-200c could suppress BMI1 expression, and suppressed proliferation and tumor formation rate (P<0.05). Compared to NTG cells, BCSCs has significantly down-regulated expression of miR-200c. BCSCs facilitate BMI1 gene expression via inhibiting miR-200c expression, further elevating BCSCs proliferation and tumor formation potency.

SUBMITTER: Wu D

PROVIDER: S-EPMC6965978 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature

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Differential expression of miR-200c in breast cancer stem cells.

Wu Dijun D Ji Ning N Zhang Lei L Zhang Liang L

International journal of clinical and experimental pathology 20170901 9

Breast cancer has become the most popular malignant tumors in females. Breast cancer stem cells (BCSCs) play an important role in metastasis and recurrence. Previous study showed close correlation between miR-200c and behavioral regulation of breast cancer stem cells. This study aimed to investigate differential expression of miR-200c in breast cancer stem cells, and its role in regulation tumor formation property. DNA microchip assay and qRT-PCR analyzed differential expression of microRNA betw ...[more]

PMID: 31966899

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Project description:Breast cancer stem cells (BCSCs) have been reported as the origin of breast cancer and the radical cause of drug resistance, relapse and metastasis in breast cancer. BCSCs could be derived from mutated mammary epithelial stem cells (MaSCs). Therefore, comparing the molecular differences between BCSCs and MaSCs may clarify the mechanism underlying breast carcinogenesis and the targets for gene therapy. Specifically, the distinct miRNome data of BCSCs and MaSCs need to be analyzed to find out the key miRNAs and reveal their roles in regulating the stemness of BCSCs.MUC1(-)ESA(+) cells were isolated from normal mammary epithelial cell line MCF-10A by fluorescence-activated cell sorting (FACS) and tested for stemness by clonogenic assay and multi-potential differentiation experiments. The miRNA profiles of MaSCs, BCSCs and breast cancer MCF-7 cells were compared to obtain the candidate miRNAs that may regulate breast tumorigenesis. An miRNA consecutively upregulated from MaSCs to BCSCs to MCF-7 cells, miR-200c, was chosen to determine its role in regulating the stemness of BCSCs and MaSCs in vitro and in vivo. Based on bioinformatics, the targets of miR-200c were validated by dual-luciferase report system, western blot and rescue experiments.In a 2-D clonogenic assay, MUC1(-)ESA(+) cells gave rise to multiple morphological colonies, including luminal colonies, myoepithelial colonies and mixed colonies. The clonogenic potential of MUC1(-)ESA(+) (61.5 ± 3.87 %) was significantly higher than that of non-stem MCF-10A cells (53.5 ± 3.42 %) (P < 0.05). In a 3-D matrigel culture, MUC1(-)ESA(+) cells grew into mammospheres with duct-like structures. A total of 12 miRNAs of interest were identified, 8 of which were upregulated and 4 downregulated in BCSCs compared with MaSCs. In gain- and lost-of-function assays, miR-200c was sufficient to inhibit the self-renewal of BCSCs and MaSCs in vitro and the growth of BCSCs in vivo. Furthermore, miR-200c negatively regulated programmed cell death 10 (PDCD10) in BCSCs and MaSCs. PDCD10 could rescue the tumorigenesis inhibited by miR-200c in BCSCs.Accumulating evidence shows that there is a milignant transformation from MaSCs into BCSCs. The underlying mechanism remains unclear. In present study, miRNA profiles between MaSCs and BCSCs were obtained. Then miRNA-200c, downregulated in both MaSCs and BCSCs, were verified as anti-oncogene, and played essential role in regulating self-renewal of both kinds of stem-like cells. These findings reveal a novel insights of breast tumorigenesis.PDCD10 is a target gene of miR-200c and also a possible mechanism by which miR-200c plays a role in regulating the stemness of BCSCs and MaSCs.

Dataset Information

Differential expression of miR-200c in breast cancer stem cells.

Publications

Differential expression of miR-200c in breast cancer stem cells.

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