网络筛选乳腺癌相关miRNAs及miR-106a-5p对乳腺癌细胞侵袭迁移能力的影响
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摘要
目的网络筛选乳腺癌关键微小RNAs(micro RNAs,miRNAs),并探讨miR-106a-5p对乳腺癌细胞侵袭及迁移能力的影响。方法通过Target Scan和miRanda等工具,构建miRNAs与靶基因以及显著靶向性功能(gene ontology,GO)的调控网络,筛选对乳腺癌有关键调控作用的miRNAs;利用miR-106a-5p agomir及miR-106a-5p antagomir转染MCF-7、T47D乳腺癌细胞株,建立乳腺癌细胞中miR-106a-5p的过表达及敲低体系,Transwell侵袭实验、Transwell迁移实验、划痕实验检测miR-106a-5p对乳腺癌细胞的侵袭、迁移能力的影响。结果在miRNAs-gene-network和miRNAs-GO-network两个网络中调控维度最高的miRNAs基本一致,其中miR-106a-5p是调控维度最高的miRNAs之一;过表达miR-106a-5p显著抑制乳腺癌细胞的侵袭能力和迁移能力(P<0.05),敲低miR-106a-5p则显著增加乳腺癌细胞的侵袭能力和迁移能力(P<0.05)。结论 miR-106a-5p是乳腺癌miRNAs调控网络中的关键miRNAs之一,具有抑制乳腺癌细胞的侵袭及迁移能力的生物学功能。
Objective To screen key miRNAs related with breast cancer and explore the modulating effect of miR-106a-5p on the invasion and migration of breast cancer cells. Methods Target Scan and miRanda softwares were employed to predict the targets of miRNA array in breast cancer cells. The miRNAsgene-network and miRNAs-GO-network were established based on gene ontology( GO) analysis. The overexpression and knockdown of miR-106a-5p were established by transfection of miR-106a-5p agomir and miR-106a-5p antagomir into MCF-7 and T47 D breast cancer cells,respectively. The invasion and migration of breast cancer cells were tested by Transwell invasion assay,Transwell migration assay and scratch assay.Results Mi R-106a-5p was one of miRNAs with the highest regulating degree in both miRNAs-gene-network and miRNAs-GO-network. The expression of miR-106a-5p in breast cancer cells was significantly lower than that in normal mammary cells( P < 0. 05). The overexpression of miR-106a-5p inhibited the invasion and migration of MCF-7 and T47 D breast cancer cells( P < 0. 05). On the contrary,the knockdown of miR-106a-5p promoted the invasion and migration of breast cancer cells( P < 0. 05). Conclusion miR-106a-5p is a key miRNA in the regulation network of breast cancer miRNAs,and can inhibit the invasion and migration of breast cancer cells.
Objective To screen key miRNAs related with breast cancer and explore the modulating effect of miR-106a-5p on the invasion and migration of breast cancer cells. Methods Target Scan and miRanda softwares were employed to predict the targets of miRNA array in breast cancer cells. The miRNAsgene-network and miRNAs-GO-network were established based on gene ontology( GO) analysis. The overexpression and knockdown of miR-106a-5p were established by transfection of miR-106a-5p agomir and miR-106a-5p antagomir into MCF-7 and T47 D breast cancer cells,respectively. The invasion and migration of breast cancer cells were tested by Transwell invasion assay,Transwell migration assay and scratch assay.Results Mi R-106a-5p was one of miRNAs with the highest regulating degree in both miRNAs-gene-network and miRNAs-GO-network. The expression of miR-106a-5p in breast cancer cells was significantly lower than that in normal mammary cells( P < 0. 05). The overexpression of miR-106a-5p inhibited the invasion and migration of MCF-7 and T47 D breast cancer cells( P < 0. 05). On the contrary,the knockdown of miR-106a-5p promoted the invasion and migration of breast cancer cells( P < 0. 05). Conclusion miR-106a-5p is a key miRNA in the regulation network of breast cancer miRNAs,and can inhibit the invasion and migration of breast cancer cells.
引文
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[17]Yang G,Zhang R,Chen X,et al.Mi R-106a inhibits glioma cell growth by targeting E2F1 independent of p53 status[J].J Mol Med(Berl),2011,89(10):1037-1050.DOI:10.1007/s00109-011-0775-x
[18]Xu L Z,Li S S,Zhou W,et al.p62/SQSTM1 enhances breast cancer stem-like properties by stabilizing MYC m RNA[J].Oncogene,2016.[Epub ahead of print].DOI:10.1038/onc.2016.202
[2]Chen W,Zheng R,Baade P D,et al.Cancer statistics in China,2015[J].CA Cancer J Clin,2016,66(2):115-132.DOI:10.3322/caac.21338
[3]Fang Y X,Gao W Q.Roles of micro RNAs during prostatic tumorigenesis and tumor progression[J].Oncogene,2014,33(2):135-147.DOI:10.1038/onc.2013.54
[4]Aydogdu E,Katchy A,Tsouko E,et al.Micro RNA-regulated gene networks during mammary cell differentiation are associated with breast cancer[J].Carcinogenesis,2012,33(8):1502-1511.DOI:10.1093/carcin/bgs161
[5]Sun J G,Liao R X,Qiu J,et al.Microarray-based analysis of micro RNA expression in breast cancer stem cells[J].J Exp Clin Cancer Res,2010,29:174.DOI:10.1186/1756-9966-29-174
[6]Bartel D P.Micro RNAs:genomics,biogenesis,mechanism,and function[J].Cell,2004,116(2):281-297.DOI:10.1016/S0092-8674(04)00045-5
[7]Lujambio A,Lowe S W.The microcosmos of cancer[J].Nature,2012,482(7385):347-355.DOI:10.1038/nature10888
[8]Cheng C J,Bahal R,Babar I A,et al.Micro RNA silencing for cancer therapy targeted to the tumour microenvironment[J].Nature,2015,518(7537):107-110.DOI:10.1038/nature13905
[9]Li L,Chen J,Chen X,et al.Serum mi RNAs as predictive and preventive biomarker for pre-clinical hepatocellular carcinoma[J].Cancer Lett,2016,373(2):234-240.DOI:10.1016/j.canlet.2016.01.028
[10]Buffa F M,Camps C,Winchester L,et al.micro RNA-associated progression pathways and potential therapeutic targets identified by integrated m RNA and micro RNA expression profiling in breast cancer[J].Cancer Res,2011,71(17):5635-5645.DOI:10.1158/0008-5472.CAN-11-0489
[11]Landais S,Landry S,Legault P,et al.Oncogenic potential of the mi R-106-363 cluster and its implication in human Tcell leukemia[J].Cancer Res,2007,67(12):5699-5707.DOI:10.1158/0008-5472.CAN-06-4478
[12]Mishra S,Srivastava A K,Suman S,et al.Circulating mi RNAs revealed as surrogate molecular signatures for the early detection of breast cancer[J].Cancer Lett,2015,369(1):67-75.DOI:10.1016/j.canlet.2015.07.045
[13]Kim K,Chadalapaka G,Lee SO,et al.Identification of oncogenic micro RNA-17-92/ZBTB4/specificity protein axis in breast cancer[J].Oncogene,2012,31(8):1034-1044.DOI:10.1038/onc.2011.296
[14]Guo X,Yang C,Qian X,et al.Estrogen receptor alpha regulates ATM Expression through mi RNAs in breast cancer[J].Clin Cancer Res,2013,19(18):4994-5002.DOI:10.1158/1078-0432.CCR-12-3700
[15]Yang W S,Chadalapaka G,Cho S G,et al.The transcriptional repressor ZBTB4 regulates EZH2 through a Micro RNA-ZBTB4-specificity protein signaling axis[J].Neoplasia,2014,16(12):1059-1069.DOI:10.1016/j.neo.2014.09.011
[16]Zhu M,Zhang N,He S,et al.Micro RNA-106a targets TIMP2 to regulate invasion and metastasis of gastric cancer[J].FEBS Lett,2014,588(4):600-607.DOI:10.1016/j.febslet.2013.12.028
[17]Yang G,Zhang R,Chen X,et al.Mi R-106a inhibits glioma cell growth by targeting E2F1 independent of p53 status[J].J Mol Med(Berl),2011,89(10):1037-1050.DOI:10.1007/s00109-011-0775-x
[18]Xu L Z,Li S S,Zhou W,et al.p62/SQSTM1 enhances breast cancer stem-like properties by stabilizing MYC m RNA[J].Oncogene,2016.[Epub ahead of print].DOI:10.1038/onc.2016.202