基础医学与临床 ›› 2016, Vol. 36 ›› Issue (4): 439-444.

• 研究论文 • 上一篇    下一篇

miR-140在骨肉瘤组织中表达降低及其过表达可促进骨肉瘤U2细胞的凋亡

陈翔1,黄路2,张中卒3,肖前仁1,陈少卿1,舒勇1,曹凯1   

  1. 1. 南昌大学第一附属医院
    2. 江西省妇幼保健院
    3. 重庆医科大学附属永川医院
  • 收稿日期:2015-09-17 修回日期:2015-11-11 出版日期:2016-04-05 发布日期:2016-03-29
  • 通讯作者: 曹凯 E-mail:kaichaw@126.com
  • 基金资助:
    从表观遗传调控角度探讨miR-140对骨肉瘤恶性表型的抑制作用;let-7a/STAT3/lin28正反馈环路对尤文肉瘤干细胞恶性表型的影响及其机制;let-7a靶向调节STAT3对尤文肉瘤干细胞恶性表型的影响及其机制;糖尿病人腰椎管狭窄症发病机制的研究;诱导多能干细胞移植对脊髓损伤致残后神经功能恢复的治疗研究

miR-140 is low expressed in osteosarcoma tissues and the over expression of miRNA-140 can promote the apoptosis of osteosarcoma cell line U2 in vitro

  • Received:2015-09-17 Revised:2015-11-11 Online:2016-04-05 Published:2016-03-29

摘要: 目的 探讨骨肉瘤组织中MiR-140的表达,以及过表达miR-140表达后对骨肉瘤细胞生物学行为的影响。方法 用实时荧光定量PCR(qRT-PCR)检测40例骨肉瘤患者肿瘤组织与癌旁组织中miR-140的表达。用脂质体法瞬时将人工合成的成熟miR-140片段(miR-140 mimic)转染至骨肉瘤U2细胞中;用qRT-PCR法检测细胞中miR-140的表达,CCK-8法及流式细胞仪检测细胞的增殖及凋亡,Transwell小室检测细胞的侵袭能力,Western印迹法检测miR-140靶基因组蛋白去乙酰化酶4(HDAC4)的表达。结果72.5%(29/40)的骨肉瘤组织miR-140表达明显低于瘤旁组织(P<0.05)。转染miR-140 mimic后U2细胞miR-140的表达水平明显上调(P<0.05),细胞增殖能力明显降低(P<0.05),凋亡率明显升高(P<0.05),侵袭能力明显下降(P<0.05),细胞中HDAC4蛋白的表达水平明显下降(P<0.05)。结论 miR-140在骨肉瘤中低表达;过表达miR-140后可抑制骨肉瘤U2细胞的增殖和侵袭,并促进其凋亡。这一作用可能部分依赖于miR-140靶向抑制HDAC4的表达。

关键词: 骨肉瘤, 微小RNA-140, 组蛋白去乙酰化酶4, 凋亡

Abstract: Objective To detect the expression of miR-140 in osteosarcoma tissues and investigate the effect of miR-140 on the proliferation, apoptosis and invasion of human osteosarcoma cell line U2. Methods The real-time polymerase chain reaction(qRT-PCR) was used to detect the expression levels of miR-140 in osteosarcoma tissues and corresponding paratumorous tissues from 40 patients. The expression levels of miR-140 in osteosarcoma cell line U2 after transfected with miR-140 mimic were measured by qRT-PCR. The proliferation and apoptosis of U2 cells were assessed by cell counting kit (CCK-8 method) and flow cytometry, respectively. And the in vitro invasion ability of U2 cells was detected with Transwell chamber assay, The protein changes of histone deacetylase 4 (HDAC4) of U2 cells after transfection were detected by Western blotting. Results The 72.5% (29/40) of miR-140 was low expressed in osteosarcoma tissues compared with the adjacent normal tissues (P<0.05).The miR-140 expression of U2 cells after transfection was significantly up-regulated (P<0.05), The apoptosis rate of U2 cells was significantly increased (P<0.05), The proliferation and invasion abilities were inhibited significantly (P<0.05). The relative protein expressions of HDAC4 in miR-140 mimic transfection group were significantly lower than those in negative control group (P<0.05). Conclusion MiR-140 is low expressed in human osteosarcoma tissue. MicroRNA-140 inhibits the proliferation and invasion activities of osteosarcoma cell line U2 and promotes cells’ apoptosis in vitro. These biological effects may partially attribute to miR-140 characters of targeting and inhibiting the HDAC4 expression.

Key words: osteosarcoma, microRNA-140, histone deacetylase 4, apoptosis