miR-200b对三阴性乳腺癌细胞MDA-MB-231的负性调控作用
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摘要
目的研究miRNA-200b对三阴性乳腺癌细胞增殖、侵袭能力的影响。方法定量PCR检测人乳腺癌细胞株MCF-7、BT-474、MDA-MB-453、SKBR3、MDA-MB-468、MDA-MB-231中miRNA-200a/b/c的表达情况,通过miR-200b抑制剂及拟似物分别抑制、促进miR-200b的表达后,观察癌细胞增殖能力、克隆形成及侵袭能力的变化。结果 miRNA-200家族在Luminal型乳腺癌细胞MCF-7、BT-474和Her-2表达型MDA-MB-453、SKBR3的表达量高于三阴性型MDA-MB-468、MDA-MB-231的表达。通过inhibitor、mimics分别抑制、促进MDA-MB-231细胞中miR-200b的表达后,结果显示miR-200b对肿瘤细胞的增殖有抑制作用(P<0.05),miRNA-200b组对肿瘤细胞的克隆形成有抑制作用(P<0.05),对癌细胞侵袭有抑制作用(P<0.05)。结论miRNA-200b对三阴性乳腺癌细胞MDA-MB-231具有负性调控作用,可以作为潜在的早期诊断及治疗靶点。
Objective To identify the regulation of miRNA-200 b on proliferation of triple negative breast cancer cells.Methods The expressions of miRNA-200 a/b/c in breast cancer cell lines MCF-7,BT-474,MDA-MB-453,SKBR3,MDA-MB-468,and MDA-MB-231 were detected by real-time PCR.MTT was used to detect MDAMB-231 cell proliferation after miR-200 b was downregulated and upregulated.The colony-forming efficiency of miR-200 b was detected by colony forming assay.The transwell chamber was used to investigate the influence of miR-200 b on invasion of MDA-MB-231 cells.Results miRNA-200 a/b/c had relatively low expressions in triple negative breast cancer cell lines MDA-MB-468 and MDA-MB-231,but high expressions in luminal cells MCF-7,BT-474,and Her-2 positive cells MDA-MB-453,SKBR3.The upregulated miR-200b could suppress the proliferation,colony formation and cell invasion of MDA-MB-231 cells(P<0.05).Also the downregulated miR-200b promoted cell proliferation,colony forming and cell invasion(P<0.05).Conclusion miRNA-200 b suppresses triple negative breast cancer cell MDA-MB-231,making it a potential biomarker and target for early diagnosis and therapy.
Objective To identify the regulation of miRNA-200 b on proliferation of triple negative breast cancer cells.Methods The expressions of miRNA-200 a/b/c in breast cancer cell lines MCF-7,BT-474,MDA-MB-453,SKBR3,MDA-MB-468,and MDA-MB-231 were detected by real-time PCR.MTT was used to detect MDAMB-231 cell proliferation after miR-200 b was downregulated and upregulated.The colony-forming efficiency of miR-200 b was detected by colony forming assay.The transwell chamber was used to investigate the influence of miR-200 b on invasion of MDA-MB-231 cells.Results miRNA-200 a/b/c had relatively low expressions in triple negative breast cancer cell lines MDA-MB-468 and MDA-MB-231,but high expressions in luminal cells MCF-7,BT-474,and Her-2 positive cells MDA-MB-453,SKBR3.The upregulated miR-200b could suppress the proliferation,colony formation and cell invasion of MDA-MB-231 cells(P<0.05).Also the downregulated miR-200b promoted cell proliferation,colony forming and cell invasion(P<0.05).Conclusion miRNA-200 b suppresses triple negative breast cancer cell MDA-MB-231,making it a potential biomarker and target for early diagnosis and therapy.
引文
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[15]刘棣,黄辰,刁冬梅,等.miRNA-449家族靶向CCNE2对胃癌细胞生长的抑制调节作用[J].西安交通大学学报(医学版),2012,33(5):544-548.
[2]CRISCITIELLO C,AZIM HA,JR,SCHOUTEN PC,et al.Understanding the biology of triple-negative breast cancer[J].Ann Oncol,2012,23(Suppl 6):vi13-18.
[3]BAYRAKTAR S,GLUCK S.Molecularly targeted therapies for metastatic triple-negative breast cancer[J].Breast Cancer Res Treat,2013,138(1):21-35.
[4]ANDRE F,ZIELINSKI CC.Optimal strategies for the treatment of metastatic triple-negative breast cancer with currently approved agents[J].Ann Oncol,2012,23(Suppl 6):vi46-51.
[5]GAO S L,WANG L Z,LIU H Y,et al.miR-200a inhibits tumor proliferation by targeting AP-2gamma inneuroblastoma cells[J].Asian Pac J Cancer Prev,2014,15(11):4671-4676.
[6]SHI L,ZHANG S,WU H,et al.MiR-200c increases theradiosensitivity of non-small-cell lung cancer cell line A549 by targeting VEGF-VEGFR2 pathway[J].PLoS One,2013,8(10):e78344.
[7]TUOMARILAM,LUOSTARI K,SOINI Y,et al.Overexpression of microRNA-200c predicts poor outcome in patients with PR-negative breast cancer[J].PLoS One,2014,9(10):e109508.
[8]MANAVALAN TT,TENG Y,LITCHFIELD LM,et al.Reduced expression of miR-200 family members contributes toantiestrogen resistance in LY2 human breast cancer cells[J].PLoS One,2013,8(4):e62334.
[9]D'IPPOLITO E,PLANTAMURA I,BONGIOVANNI L,et al.miR-9 and miR-200 regulatePDGFRbeta-mediated endothelial differentiation of tumor cells in triple-negative breast cancer[J].Cancer Res,2016,76(18):5562-5572.
[10]PERDIGAO-HENRIQUES R,PETROCCA F,ALTSCHULER G,et al.miR-200 promotes themesenchymal to epithelial transition by suppressing multiple members of the Zeb2 and Snail1 transcriptional repressor complexes[J].Oncogene,2016,35(2):158-172.
[11]KORPAL M,ELL BJ,BUFFA FM,et al.Direct targeting of Sec23a by miR-200s influences cancer cellsecretome and promotes metastatic colonization[J].Nat Med,2011,17(9):1101-1108.
[12]CHOI SK,KIM HS,JIN T,et al.vOverexpression of the miR-141/200c cluster promotes the migratory and invasive ability of triple-negative breast cancer cells through the activation of the FAK and PI3K/AKT signaling pathways by secreting VEGF-A[J].BMC Cancer,2016,16:570.
[13]HUMPHRIES B,WANG Z,OOM AL,et al.MicroRNA-200b targets proteinkinase Calpha and suppresses triple-negative breast cancer metastasis[J].Carcinogenesis,2014,35(10):2254-2263.
[14]LIU T,ZHANG X,SHANG M,et al.Dysregulated expression of Slug,vimentin,and E-cadherin correlates with poor clinical outcome in patients with basal-like breast cancer[J].J Surg Oncol,2013,107(2):188-194.
[15]刘棣,黄辰,刁冬梅,等.miRNA-449家族靶向CCNE2对胃癌细胞生长的抑制调节作用[J].西安交通大学学报(医学版),2012,33(5):544-548.