微小RNA-10b调控乳腺癌MCF-7细胞中乳腺癌干细胞标记物乙醛脱氢酶1 mRNA和蛋白的表达
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摘要
背景:miR-10b能够调控乳腺癌干细胞的相关特性,乙醛脱氢酶1是最重要的乳腺癌干细胞标记物之一,二者在乳腺癌细胞中的相互作用尚需进一步研究。目的:探讨miR-10b对乳腺癌MCF-7细胞中乳腺癌干细胞标记物乙醛脱氢酶1 mRNA和蛋白表达的影响。方法:常规培养乳腺癌MCF-7细胞株,分别将miR-10b模拟物或其阴性对照转染进入乳腺癌MCF-7细胞;转染48 h,通过实时荧光定量PCR实验和Western blot实验检测乙醛脱氢酶1 mRNA和蛋白的表达。结果与结论:(1)将miR-10b模拟物转染进入MCF-7细胞之后,miR-10b在MCF-7细胞株中的表达显著高于对照组(P=0.003 47);(2)miR-10b过表达之后显著上调了MCF-7细胞株中乙醛脱氢酶1 mRNA和蛋白的表达(P=0.009 54和P=0.003 11);(3)结果表明,上调miR-10b表达能够诱导乙醛脱氢酶1阳性乳腺癌干细胞的产生,从而为miR-10b调控乳腺癌细胞侵袭、转移提供新的研究依据。
BACKGROUND: MicroR-10 b can regulate the characteristics of breast cancer stem cells, and acetaldehyde dehydrogenase 1(ALDH1) is one of the most important markers of breast cancer stem cells. The interaction between them in breast cancer cells needs further explorations. OBJECTIVE: To investigate whether over-expression of microRNA-10 b affects ALDH1 mRNA and protein levels in human breast cancer MCF-7 cells. METHODS: hsa-miR-10 b mimics or its negative control was transfected into breast cancer MCF-7 cell line. At 48 hours after transfection, the mRNA and protein expression of ALDH1 in the cells was detected using real-time RT-PCR and western blot assays, respectively. RESULTS AND CONCLUSION: Over-expression of microR-10 b was found in the MCF-7 cell line transfected with hsa-miR-10 b mimics, which was significantly higher than that in the control group(P=0.003 47). Both of ALDH1 mRNA and protein levels were up-regulated in the MCF-7 cell line overexpressing microR-10 b, as compared with the control group(P=0.009 54 and P=0.003 11, respectively). To conclude, over-expression of microR-10 b induces the ALDH1 mRNA and protein expression in the breast cancer MCF-7 cell line, providing new evidence that microR-10 b regulates the invasion and metastasis of breast cancer cells.
BACKGROUND: MicroR-10 b can regulate the characteristics of breast cancer stem cells, and acetaldehyde dehydrogenase 1(ALDH1) is one of the most important markers of breast cancer stem cells. The interaction between them in breast cancer cells needs further explorations. OBJECTIVE: To investigate whether over-expression of microRNA-10 b affects ALDH1 mRNA and protein levels in human breast cancer MCF-7 cells. METHODS: hsa-miR-10 b mimics or its negative control was transfected into breast cancer MCF-7 cell line. At 48 hours after transfection, the mRNA and protein expression of ALDH1 in the cells was detected using real-time RT-PCR and western blot assays, respectively. RESULTS AND CONCLUSION: Over-expression of microR-10 b was found in the MCF-7 cell line transfected with hsa-miR-10 b mimics, which was significantly higher than that in the control group(P=0.003 47). Both of ALDH1 mRNA and protein levels were up-regulated in the MCF-7 cell line overexpressing microR-10 b, as compared with the control group(P=0.009 54 and P=0.003 11, respectively). To conclude, over-expression of microR-10 b induces the ALDH1 mRNA and protein expression in the breast cancer MCF-7 cell line, providing new evidence that microR-10 b regulates the invasion and metastasis of breast cancer cells.
引文
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[8]Liu X, Guan Y, Wang L, et al. MicroRNA-10b expression in node-negative breast cancer-correlation with metastasis and angiogenesis. Oncol Lett. 2017;14(5):5845-5852.
[9]Huang Q, Song Q, Zhong W, et al. MicroRNA-10b and the clinical outcomes of various cancers:A systematic review and meta-analysis. Clin Chim Acta. 2017;474:14-22.
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[11]Yanwirasti, A Harahap W, Arisanty D. Evaluation of MiR-21and MiR-10b Expression of Human Breast Cancer in West Sumatera. Pak J Biol Sci. 2017;20(4):189-196.
[12]Ma L. Role of miR-10b in breast cancer metastasis. Breast Cancer Res. 2010;12(5):210.
[13]Bahena-Ocampo I, Espinosa M, Ceballos-Cancino G, et al.miR-10b expression in breast cancer stem cells supports self-renewal through negative PTEN regulation and sustained AKT activation. EMBO Rep. 2016;17(5):648-658.
[14]Pindiprolu SKSS, Krishnamurthy PT, Chintamaneni PK.Pharmacological targets of breast cancer stem cells:a review.Naunyn Schmiedebergs Arch Pharmacol. 2018;391(5):463-479.
[15]Yang F, Xu J, Tang L, et al. Breast cancer stem cell:the roles and therapeutic implications. Cell Mol Life Sci. 2017;74(6):951-966.
[16]Ginestier C, Hur MH, Charafe-Jauffret E, et al. ALDH1 is a marker of normal and malignant human mammary stem cells and a predictor of poor clinical outcome. Cell Stem Cell. 2007;1(5):555-567.
[17]Louhichi T, Ziadi S, Saad H, et al. Clinicopathological significance of cancer stem cell markers CD44 and ALDH1expression in breast cancer. Breast Cancer. 2018;25(6):698-705.
[18]Kong Y, Lyu N, Wu J, et al. Breast cancer stem cell markers CD44 and ALDH1A1 in serum:distribution and prognostic value in patients with primary breast cancer. J Cancer. 2018;9(20):3728-3735.
[19]Mandujano-Tinoco EA, García-Venzor A, Melendez-Zajgla J,et al. New emerging roles of microRNAs in breast cancer.Breast Cancer Res Treat. 2018;171(2):247-259.
[20]Mehrgou A, Akouchekian M. Therapeutic impacts of microRNAs in breast cancer by their roles in regulating processes involved in this disease. J Res Med Sci. 2017;22:130.
[21]Wang P, Liu XM, Ding L, et al. mTOR signaling-related MicroRNAs and Cancer involvement. J Cancer. 2018;9(4):667-673.
[22]Weidle UH, Dickopf S, Hintermair C, et al. The Role of micro RNAs in Breast Cancer Metastasis:Preclinical Validation and Potential Therapeutic Targets. Cancer Genomics Proteomics.2018;15(1):17-39.
[23]Chen W, Hoffmann AD, Liu H, et al. Organotropism:new insights into molecular mechanisms of breast cancer metastasis. NPJ Precis Oncol. 2018;2(1):4.
[24]Han M, Wang F, Gu Y, et al. MicroRNA-21 induces breast cancer cell invasion and migration by suppressing smad7 via EGF and TGF-βpathways. Oncol Rep. 2016;35(1):73-80.
[25]Han M, Liu M, Wang Y, et al. Antagonism of miR-21 reverses epithelial-mesenchymal transition and cancer stem cell phenotype through AKT/ERK1/2 inactivation by targeting PTEN. PLoS One. 2012;7(6):e39520.
[26]Han M, Wang Y, Liu M, et al. MiR-21 regulates epithelial-mesenchymal transition phenotype and hypoxia-inducible factor-1αexpression in third-sphere forming breast cancer stem cell-like cells. Cancer Sci. 2012;103(6):1058-1064.
[27]Han M, Liu M, Wang Y, et al. Re-expression of miR-21contributes to migration and invasion by inducing epithelial-mesenchymal transition consistent with cancer stem cell characteristics in MCF-7 cells. Mol Cell Biochem. 2012;363(1-2):427-436.
[28]Han ML, Wang F, Gu YT, et al. MicroR-760 suppresses cancer stem cell subpopulation and breast cancer cell proliferation and metastasis:By down-regulating NANOG.Biomed Pharmacother. 2016;80:304-310.
[29]Han M, Wang Y, Guo G, et al. microRNA-30d mediated breast cancer invasion, migration, and EMT by targeting KLF11 and activating STAT3 pathway. J Cell Biochem. 2018;119(10):8138-8145.
[2]Bertoli G, Cava C, Castiglioni I. MicroRNAs:New Biomarkers for Diagnosis, Prognosis, Therapy Prediction and Therapeutic Tools for Breast Cancer. Theranostics. 2015;5(10):1122-1143.
[3]林庆铿,尹德宏,刘菊芬,等.第2代和第5代人脐带间充质干细胞外泌体miRNA谱的比较[J].中国组织工程研究,2018,22(17):2686-2691.
[4]Mytsyk Y, Dosenko V, Skrzypczyk MA, et al. Potential clinical applications of microRNAs as biomarkers for renal cell carcinoma. Cent European J Urol. 2018;71(3):295-303.
[5]Link A, Kupcinskas J. MicroRNAs as non-invasive diagnostic biomarkers for gastric cancer:Current insights and future perspectives. World J Gastroenterol. 2018;24(30):3313-3329.
[6]Huang Q, Song Q, Zhong W, et al. MicroRNA-10b and the clinical outcomes of various cancers:A systematic review and meta-analysis. Clin Chim Acta. 2017;474:14-22.
[7]Biagioni F, Bossel Ben-Moshe N, Fontemaggi G, et al. The locus of microRNA-10b:a critical target for breast cancer insurgence and dissemination. Cell Cycle. 2013;12(15):2371-2375.
[8]Liu X, Guan Y, Wang L, et al. MicroRNA-10b expression in node-negative breast cancer-correlation with metastasis and angiogenesis. Oncol Lett. 2017;14(5):5845-5852.
[9]Huang Q, Song Q, Zhong W, et al. MicroRNA-10b and the clinical outcomes of various cancers:A systematic review and meta-analysis. Clin Chim Acta. 2017;474:14-22.
[10]顾忆,李小曼,郑禄枫,等. MicroRNA-10b调控乳腺癌细胞生长增殖与凋亡的研究[J].中国药科大学学报, 2015, 46(2):242-249.
[11]Yanwirasti, A Harahap W, Arisanty D. Evaluation of MiR-21and MiR-10b Expression of Human Breast Cancer in West Sumatera. Pak J Biol Sci. 2017;20(4):189-196.
[12]Ma L. Role of miR-10b in breast cancer metastasis. Breast Cancer Res. 2010;12(5):210.
[13]Bahena-Ocampo I, Espinosa M, Ceballos-Cancino G, et al.miR-10b expression in breast cancer stem cells supports self-renewal through negative PTEN regulation and sustained AKT activation. EMBO Rep. 2016;17(5):648-658.
[14]Pindiprolu SKSS, Krishnamurthy PT, Chintamaneni PK.Pharmacological targets of breast cancer stem cells:a review.Naunyn Schmiedebergs Arch Pharmacol. 2018;391(5):463-479.
[15]Yang F, Xu J, Tang L, et al. Breast cancer stem cell:the roles and therapeutic implications. Cell Mol Life Sci. 2017;74(6):951-966.
[16]Ginestier C, Hur MH, Charafe-Jauffret E, et al. ALDH1 is a marker of normal and malignant human mammary stem cells and a predictor of poor clinical outcome. Cell Stem Cell. 2007;1(5):555-567.
[17]Louhichi T, Ziadi S, Saad H, et al. Clinicopathological significance of cancer stem cell markers CD44 and ALDH1expression in breast cancer. Breast Cancer. 2018;25(6):698-705.
[18]Kong Y, Lyu N, Wu J, et al. Breast cancer stem cell markers CD44 and ALDH1A1 in serum:distribution and prognostic value in patients with primary breast cancer. J Cancer. 2018;9(20):3728-3735.
[19]Mandujano-Tinoco EA, García-Venzor A, Melendez-Zajgla J,et al. New emerging roles of microRNAs in breast cancer.Breast Cancer Res Treat. 2018;171(2):247-259.
[20]Mehrgou A, Akouchekian M. Therapeutic impacts of microRNAs in breast cancer by their roles in regulating processes involved in this disease. J Res Med Sci. 2017;22:130.
[21]Wang P, Liu XM, Ding L, et al. mTOR signaling-related MicroRNAs and Cancer involvement. J Cancer. 2018;9(4):667-673.
[22]Weidle UH, Dickopf S, Hintermair C, et al. The Role of micro RNAs in Breast Cancer Metastasis:Preclinical Validation and Potential Therapeutic Targets. Cancer Genomics Proteomics.2018;15(1):17-39.
[23]Chen W, Hoffmann AD, Liu H, et al. Organotropism:new insights into molecular mechanisms of breast cancer metastasis. NPJ Precis Oncol. 2018;2(1):4.
[24]Han M, Wang F, Gu Y, et al. MicroRNA-21 induces breast cancer cell invasion and migration by suppressing smad7 via EGF and TGF-βpathways. Oncol Rep. 2016;35(1):73-80.
[25]Han M, Liu M, Wang Y, et al. Antagonism of miR-21 reverses epithelial-mesenchymal transition and cancer stem cell phenotype through AKT/ERK1/2 inactivation by targeting PTEN. PLoS One. 2012;7(6):e39520.
[26]Han M, Wang Y, Liu M, et al. MiR-21 regulates epithelial-mesenchymal transition phenotype and hypoxia-inducible factor-1αexpression in third-sphere forming breast cancer stem cell-like cells. Cancer Sci. 2012;103(6):1058-1064.
[27]Han M, Liu M, Wang Y, et al. Re-expression of miR-21contributes to migration and invasion by inducing epithelial-mesenchymal transition consistent with cancer stem cell characteristics in MCF-7 cells. Mol Cell Biochem. 2012;363(1-2):427-436.
[28]Han ML, Wang F, Gu YT, et al. MicroR-760 suppresses cancer stem cell subpopulation and breast cancer cell proliferation and metastasis:By down-regulating NANOG.Biomed Pharmacother. 2016;80:304-310.
[29]Han M, Wang Y, Guo G, et al. microRNA-30d mediated breast cancer invasion, migration, and EMT by targeting KLF11 and activating STAT3 pathway. J Cell Biochem. 2018;119(10):8138-8145.