[1]胡欣春,魏益平,彭金华,等.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,et al.Regulation of miR-4686 on proliferation in A549 spheres[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(04):340-345.
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miR-4686对A549细胞球增殖的调控作用及其机制研究(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
36卷
期数:
2014年第04期
页码:
340-345
栏目:
论著
出版日期:
2014-02-28

文章信息/Info

Title:
Regulation of miR-4686 on proliferation in A549 spheres
作者:
胡欣春魏益平彭金华喻东亮徐建军
南昌大学第二附属医院心胸外科;南昌大学医学院;江西省胸科医院胸外科
Author(s):
Hu Xinchun Wei Yiping Peng Jinhua Yu Dongliang Xu Jianjun
Department of Cardiothoracic Surgery, Second Affiliated Hospital, Nanchang University, Nanchang, Jiangxi Province, 330006; Medicine School of Nanchang University, Nanchang, Jiangxi Province, 330006; Department of Chest Surgery, Jiangxi Chest Hospital, Nanchang, Jiangxi Province, 330006, China
关键词:
microRNA 抑制物CD133肺癌
Keywords:
microRNA inhibitors CD133 lung cancer
分类号:
R73-362;R730.23;R734.2
文献标志码:
A
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(P<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(P<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 control 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 (P<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 (P<0.05). What’s more, OD values at 12, 24 and 48 h were decreased after miR-4686 inhibitor treatment (P<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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更新日期/Last Update: 2014-02-12