miRNA-301b对三阴性型乳腺癌的血管生成拟态和预后的影响

Effect of miRNA-301b on vasculogenic mimicry and prognosis of triple-negative breast cancer in vitro

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(P75)]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.