miR-379-5p抑制小鼠乳腺癌4T1细胞的增殖、侵袭和迁移

MiR-379-5p inhibits proliferation,invasion,and migration of mouse breast cancer 4T1 cells

目的 通过研究miR-379-5p对小鼠乳腺癌4T1细胞增殖、侵袭和迁移的影响,为临床抑制乳腺癌增殖、侵袭和转移提供新的治疗靶点。方法 采用实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction, qRT-PCR)检测miR-379-5p在质粒转染后4T1细胞中的表达情况;采用5-乙炔基-2’脱氧尿嘧啶核苷(5-ethynyl-2’-deoxyuridine, EdU)细胞增殖实验与Transwell实验检测各组4T1细胞增殖能力与侵袭能力;然后通过伤口愈合实验来检测过表达以及敲低miR-379-5p对4T1细胞迁移能力的影响;利用BABL/c小鼠建立乳腺癌小鼠移植瘤模型,观察miR-379-5p过表达后对体内肿瘤生长状况,肺转移灶数量及大小的影响。结果 EdU结果显示,与对照组相比,敲低miR-379-5p能增强乳腺癌细胞的增殖能力,过表达miR-379-5p能够抑制乳腺癌细胞的增殖能力(P<0.05);Transwell与伤口愈合实验结果显示,敲低miR-379-5p能够增强乳腺癌细胞的侵袭和迁移能力,过表达miR-379-5p显著抑制细胞的侵袭和迁移能力(P<0.01)。BABL/c小鼠体内成瘤实验显示,过表达miR-379-5p可显著减缓肿瘤的生长速度(P<0.05),对肺转移有抑制作用(P<0.01)。结论 miR-379-5p在乳腺癌中起到抑癌基因的作用,抑制小鼠乳腺癌4T1细胞的增殖、侵袭和迁移。

Objective By investigating the effects of miR-379-5p on the proliferation,invasion and metastasis of mouse breast cancer 4T1 cells,we aimed to provide new therapeutic targets for the clinical inhibition of breast cancer proliferation,invasion,and metastasis.Methods After plasmid transfection,4T1 cells were utilized to detect the expression of miR-379-5p using fluorescence quantitative PCR.While 5-ethynyl-2’doxyuridine(EdU)cell proliferation and Transwell assays were employed to detect changes in the proliferation and invasion ability of 4T1 cells in each group.The migration ability of 4T1 cells after overexpression and knockdown of miR-379-5p was examined by scratch healing assay.A transplanted tumor model of breast cancer was established in BABL/c mice,and the effects of overexpressing miR-379-5p on tumor growth and the number and size of lung metastases were observed.Results EdU result showed that knocking down miR-379-5p enhanced the proliferation ability of the cells compared with the control group cells,and miR-379-5p overexpression reduced the capacity of breast cancer cells to proliferate(P<0.05).Transwell and wound healing assays showed that miR-379-5p knockdown enhanced,while miR-379-5p overexpression significantly inhibited,the invasion and migratory ability of breast cancer cells(P<0.01).An in vivo tumorigenesis experiment with BABL/c mice showed that miR-379-5p overexpression significantly slowed the tumor growth rate(P<0.05)and inhibited lung metastasis(P<0.01).Conclusions MiR-379-5p plays a role in tumor gene suppression in breast cancer and inhibits the proliferation,invasion,and migration of mouse breast cancer 4T1 cells.