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  

miR-140 is low expressed in osteosarcoma tissues and its over-expression promotes the apoptosis of osteosarcoma cell line U2 in vitro

作　　者：陈翔[1] 黄路[2] 张中卒[3] 肖前仁 陈少卿[4] 舒勇[1] 曹凯[1] 

机构地区：[1]南昌大学第一附属医院骨科,江西南昌330006 [2]江西省妇幼保健院儿童保健科,江西南昌330006 [3]重庆医科大学附属永川医院骨科,重庆402160 [4]南昌大学第一附属医院肿瘤科,江西南昌330006

出　　处：《基础医学与临床》2016年第4期439-444,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金(81260399;81460405);江西省自然科学基金(20152ACB21024;20133BCB23027);江西省科技计划支撑项目(20132BBG70068)

摘　　要：目的探讨骨肉瘤组织中miR-140的表达,以及过表达miR-140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。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 qRT-PCR was used to detect the expression level of miR-140 in osteosarcoma tissues and corresponding paratumorous tissues collected from 40 patients. The expression level of miR-140 in osteosarcoma cell line U2 after transfected with miR-140 mimic was measured by qRT-PCR. The proliferation and apoptosis of U2 cells were assessed by cell countingkit（ CCK-8 method） and flow cytometry,respectively. And the in vitro invasion ability of U2 cells was detected with Transwell chamber assay,The protein change of histone deacetylase 4（ HDAC4） of U2 cells after transfection was detected by Western blotting. Results 72. 5%（ 29 /40） of miR-140 was low expressed in osteosarcoma tissues compared with the adjacent normal tissues（ P〈0. 05）. miR-140 expression of U2 cells after transfection was significantly up-regulated（ P〈0. 05）,The apoptosis rate of U2 cells was increased significantly（ P〈0. 05）,The proliferation and invasion abilities were inhibited significantly（ P〈0. 05）. The relative protein expression of HDAC4 in miR- 140 mimic transfection group was significantly lower than those in negative control group（ P〈0. 05）. Conclusions 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.

关 键 词：骨肉瘤 微小RNA-140 组蛋白去乙酰化酶4 凋亡 

分 类 号：R738.1[医药卫生—肿瘤]