miRNA-301b对三阴性型乳腺癌的血管生成拟态和预后的影响
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摘要
目的探讨微小核糖核酸301b(miR-301b)对体外三阴性型乳腺癌(TNBC)细胞血管生成拟态的作用,并结合微阵列数据库的生物信息学数据分析miR-301b对TNBC患者预后的影响。方法体外培养TNBC细胞株MDA-MB-231细胞,使用脂质体Lipofectamine2000将miR-301b模拟物(miRNA过表达组)、miR-301b抑制剂(miRNA抑制组)、miR-301b阴性对照(阴性对照组)分别转染于MDA-MB-231细胞,通过血管生成拟态实验验证miR-301b在TNBC细胞中的血管生成作用。同时,通过Kaplan-Meier Plotter平台(http://kmplot.com/analysis/)的微阵列数据库获取相关数据分析miR-301b高/低表达对TNBC患者10年、15年生存的影响。结果血管生成拟态实验表明,miR-301b过表达组MDA-MB-231细胞的管形成数量分别是阴性对照组的3.9倍(P<0.01)、miR-301b抑制组的9.6倍(P<0.01),而miR-301b抑制组MDA-MB-231细胞的管形成数量比阴性对照组减少了58%(P<0.01)。根据微阵列数据库获取相关的10年、15年的队列随访数据,miR-301b的高表达是TNBC患者不良预后的危险因素(HR=2.09,95%CI:1.21～3.60,P=0.0068);第3四分位数(P_(75))生存时间:miR-301b高表达队列TNBC患者是38.27个月,miR-301b低表达队列TNBC患者是106.59个月。结论 MDA-MB-231细胞中存在血管拟态现象,miR-301b过表达促进TNBC细胞的血管生成,是TNBC的一个负性预后因子。进一步寻找、研究、验证miR-301在TNBC中的靶基因,有望成为TNBC抗血管生成治疗的一个突破口。
Objective To investigate the effect of microRNA(miR)-301b on vasculogenic mimicry of triple-negative breast cancer(TNBC) cells in vitro,and to analyze the prognostic effect of miR-301b on TNBC patients by bioinformatics data from microarray database.Methods TNBC cell line MDA-MB-231 cells were cultured in vitro.Lipofectamine 2000 was used to transfect miR-301b mimic(miR-301b overexpression group),miR-301b inhibitor(miR-301b inhibition group)and miR-301b negative control(NC group)into MDA-MB-231 cells,respectively.Vasculogenic mimicry experiment was used to verify the effect of angiogenesis of miR-301b in MDA-MB-231 cells.At the same time,relevant data were obtained from the microarray database of the Kaplan-Meier Plotter platform(http://kmplot.com/analysis/) to analyze the effect of miR-301b high/low expression on the survival of TNBC patients at 10 and 15 years.Results Vasculogenic mimicry experiment showed that the number of tube formation of MDA-MB-231 cells in miR-301b overexpression group was 3.9 times as much as that in NC group(P<0.01) and 9.6 times as much as that in miR-301b inhibition group(P<0.01),respectively,while the number of tube formation in miR-301b inhibition group was reduced by 58% compared with NC group(P<0.01).According to followed-up cohort data of 10 years and 15 years from microarray database,high expression of miR-301b was a risk factor of poor prognosis in patients with TNBC(HR=2.09,95%CI:1.21-3.60,P=0.0068).The survival time [third four quantile(P_(75))] was 38.27 months for TNBC patients with miR-301b high-expression cohort,and was 106.59 months for TNBC patients with miR-301b low-expression cohort.Conclusions Vasculogenic mimicry exists in MDA-MB-231 cells.Overexpression of miR-301b can promote angiogenesis of TNBC cells and is a negative prognostic factor of TNBC.Further searching,studying and verifying the target gene of miR-301b in TNBC is expected to be a breakthrough in anti-angiogenesis therapy of TNBC.
Objective To investigate the effect of microRNA(miR)-301b on vasculogenic mimicry of triple-negative breast cancer(TNBC) cells in vitro,and to analyze the prognostic effect of miR-301b on TNBC patients by bioinformatics data from microarray database.Methods TNBC cell line MDA-MB-231 cells were cultured in vitro.Lipofectamine 2000 was used to transfect miR-301b mimic(miR-301b overexpression group),miR-301b inhibitor(miR-301b inhibition group)and miR-301b negative control(NC group)into MDA-MB-231 cells,respectively.Vasculogenic mimicry experiment was used to verify the effect of angiogenesis of miR-301b in MDA-MB-231 cells.At the same time,relevant data were obtained from the microarray database of the Kaplan-Meier Plotter platform(http://kmplot.com/analysis/) to analyze the effect of miR-301b high/low expression on the survival of TNBC patients at 10 and 15 years.Results Vasculogenic mimicry experiment showed that the number of tube formation of MDA-MB-231 cells in miR-301b overexpression group was 3.9 times as much as that in NC group(P<0.01) and 9.6 times as much as that in miR-301b inhibition group(P<0.01),respectively,while the number of tube formation in miR-301b inhibition group was reduced by 58% compared with NC group(P<0.01).According to followed-up cohort data of 10 years and 15 years from microarray database,high expression of miR-301b was a risk factor of poor prognosis in patients with TNBC(HR=2.09,95%CI:1.21-3.60,P=0.0068).The survival time [third four quantile(P_(75))] was 38.27 months for TNBC patients with miR-301b high-expression cohort,and was 106.59 months for TNBC patients with miR-301b low-expression cohort.Conclusions Vasculogenic mimicry exists in MDA-MB-231 cells.Overexpression of miR-301b can promote angiogenesis of TNBC cells and is a negative prognostic factor of TNBC.Further searching,studying and verifying the target gene of miR-301b in TNBC is expected to be a breakthrough in anti-angiogenesis therapy of TNBC.
引文
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[2] Kang Y,Siegel PM,Shu W,et al.A multigenic program mediating breast cancer metastasis to bone[J].Cancer Cell,2003,3(6):537-549.
[3] Feys L,Descamps B,Vanhove C,et al.Radiation-induced lung damage promotes breast cancer lung-metastasis through CXCR4 signaling[J].Oncotarget,2015,6(29):26615-26632.
[4] Goldhirsch A,Wood WC,Coates AS,et al.Strategies for subtypes-dealing with the diversity of breast cancer:highlights of the St.Gallen International Expert Consensus on the Primary Therapy of Early Breast Cancer 2011[J].Ann Oncol,2011,22(8):1736-1747.
[5] Karlsson E,Appelgren J,Solterbeck A,et al.Breast cancer during follow-up and progression-A population based cohort on new cancers and changed biology[J].Eur J Cancer,2014,50(17):2916-2924.
[6] Neophytou C,Boutsikos P,Papageorgis P.Molecular mechanisms and emerging therapeutic targets of triple-negative breast cancer metastasis[J].Front Oncol,2018,8:31.
[7] Lee EJ,Gusev Y,Jiang J,et al.Expression profiling identifies microRNA signature in pancreatic cancer[J].Int J Cancer,2007,120(5):1046-1054.
[8] Jiang J,Gusev Y,Aderca I,et al.Association of microRNA expression in hepatocellular carcinomas with hepatitis infection,cirrhosis,and patient survival[J].Clin Cancer Res,2008,14(2):419-427.
[9] Miko E,Czimmerer Z,Csánky E,et al.Differentially expressed microRNAs in small cell lung cancer[J].Exp Lung Res,2009,35(8):646-664.
[10] Lu YC,Chen YJ,Wang HM,et al.Oncogenic function and early detection potential of miRNA-10b in oral cancer as identified by microRNA profiling[J].Cancer Prev Res (Phila),2012,5(4):665-674.
[11] Wang YX,Zhang XY,Zhang BF,et al.Initial study of microRNA expression profiles of colonic cancer without lymph node metastasis[J].J Dig Dis,2010,11(1):50-54.
[12] Song H,Li D,Wu T,et al.MicroRNA-301b promotes cell proliferation and apoptosis resistance in triple-negative breast cancer by targeting CYLD[J].BMB Rep,2018,51(11):602-607.
[13] Lánczky A,Nagy á,Bottai G,et al.miRpower:a web-tool to validate survival-associated miRNAs utilizing expression data from 2178 breast cancer patients[J].Breast Cancer Res Treat,2016,160(3):439-446.
[14] Cao G,Huang B,Liu Z,et al.Intronic miR-301 feedback regulates its host gene,ska2,in A549 cells by targeting MEOX2 to affect ERK/CREB pathways[J].Biochem Biophys Res Commun,2010,396(4):978-982.
[15] Shi W,Gerster K,Alajez NM,et al.MicroRNA-301 mediates proliferation and invasion in human breast cancer[J].Cancer Res,2011,71(8):2926-2937.
[16] Luan YY,Liu ZM,Zhong JY,et al.Effect of grape seed proanthocyanidins on tumor vasculogenic mimicry in human triple-negative breast cancer cells[J].Asian Pac J Cancer Prev,2015,16(2):531-535.
[17] Plantamura I,Casalini P,Dugnani E,et al.PDGFRβ and FGFR2 mediate endothelial cell differentiation capability of triple negative breast carcinoma cells[J].Mol Oncol,2014,8(5):968-981.