Regulation of miR-4686 on proliferation in A549 spheres

胡欣春; 魏益平; 彭金华; 喻东亮; 徐建军

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Regulation of miR-4686 on proliferation in A549 spheres

更新时间：2026-01-09

- Regulation of miR-4686 on proliferation in A549 spheres
- Journal of Army Medical University Vol. 36, Issue 4, Pages: 340-345(2014)
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  CLC： R73-362;R730.23;R734.2
- Published：2014
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胡欣春,魏益平,彭金华,喻东亮,徐建军.miR-4686对A549细胞球增殖的调控作用及其机制研究[J].陆军军医大学学报(原第三军医大学学报),2014,36(04):340-345.

　Hu Xinchun,Wei Yiping,Peng Jinhua,Yu Dongliang,Xu Jianjun.Regulation of miR-4686 on proliferation in A549 spheres[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(04):340-345.
胡欣春,魏益平,彭金华,喻东亮,徐建军.miR-4686对A549细胞球增殖的调控作用及其机制研究[J].陆军军医大学学报(原第三军医大学学报),2014,36(04):340-345. DOI：

　Hu Xinchun,Wei Yiping,Peng Jinhua,Yu Dongliang,Xu Jianjun.Regulation of miR-4686 on proliferation in A549 spheres[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(04):340-345. DOI：

摘要

目的研究miR-4686对A549细胞球增殖的调控作用及其相关机制。方法培养A549细胞，从中分选出CD133

CD44

A549细胞并置于无血清培养基中培养，形成细胞球后进行CD133

CD44

表达检测。Western blot检测A549细胞和A549细胞球中Wnt5a、Wnt2蛋白的表达；合成miR-4686抑制物和阴性对照，分别转染A549细胞球48 h后，应用Real-time PCR检测转染前后miR-4686的表达变化后，使用Real-time PCR和Western blot分析Wnt5a、Wnt2以及β-catenin mRNA和蛋白的表达变化，利用MTT检测12、24、48 h细胞的增殖能力。结果流式细胞术检测A549细胞球CD133

CD44

表达为69.800%；A549细胞球分别转染miR-4686抑制物和阴性对照物后，miR-4686的表达以及Wnt5a、Wnt2和β-catenin mRNA和蛋白的表达较对照组和miR-4686阴性对照物组显著下调(

0.05）；A549细胞球转染miR-4686抑制物后12、24、48 h，其OD值显著低于对照组和miR-4686对照物干预组（

0.05）。结论 miR-4686介导Wnt信号通路发挥调控A549细胞球增殖的作用，其可成为治疗肺癌的潜在靶点。

Abstract

Objective To study the mechanism of miR-4686 regulating the proliferation of A549 spheres. Methods CD133

CD44

A549 cells were sorted by FACS

and then cultured in serum-free medium. Positive rate of CD133

CD44

A549 spheres were detected by FACS. Wnt5a and Wnt2 protein expression of A549 cells and A549 spheres were assayed by Western blotting. MiR-4686 inhibitors and negative co

ntrol were synthesized. A549 spheres were treated with miR-4686 inhibitors and negative control separately for 48 h

and then the expression of miR-4686 were detected by real-time PCR. Wnt5a

Wnt2 and β-catenin mRNA and protein expression was assayed by RT-PCR and Western blotting

respectively. The proliferation of A549 spheres was detected by MTT assay at 12

24 and 48 h. Results CD133

CD44

A549 cells were sorted. The positive rate of CD133

CD44

was 69.800% in A549 spheres. Wnt5a and Wnt2 protein expression was significant decreased in A549 cells

as compared to A549 spheres (

0.05). After treatment with miR-4686 inhibitors

the expression of miR-4686 as well as the mRNA and protein expression of Wnt5a

Wnt2 and β-catenin were significant down-regulated

compared to the control group and miR-4686 negative control group (P0.05). What’s more

OD values at 12

24 and 48 h were decreased after miR-4686 inhibitor treatment (

0.05). Conclusion MiR-4686 plays an important role in regulation of A549 sphere proliferation through Wnt signal pathway

which may become a novel target for treatment of non-small-cell lung cancer.

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