摘要
目的:分析MicroRNA-93(miR-93)在骨肉瘤组织中的表达,探讨其对骨肉瘤细胞的增殖及细胞周期的影响。方法:采用TagManMGB探针法定量分析38例骨肉瘤组织及对应软骨组织中miR-93的表达;构建miR-93过表达载体,并合成miR-93的反义核苷酸序列,分别转染骨肉瘤细胞U2-OS和MG-63。采用MTT比色法检测骨肉瘤细胞增殖的改变情况,利用流式细胞仪检测骨肉瘤细胞周期的变化情况。结果:在38例骨肉瘤标本中,65.8%(25/38)的骨肉瘤组织miR-93表达明显高于对应软骨组织(P<0.05)。miR-93过表达载体可以促进骨肉瘤细胞的生长,使细胞周期G0/G1期明显降低,而S期比例明显增加;反义miR-93转染骨肉瘤细胞U2-OS和MG-63后,miR-93的表达明显降低,U2-OS和MG-63骨肉瘤细胞生长受到明显抑制,其生长主要停滞在G0/G1期,而S期和G2/M期细胞的比例下降。结论:miR-93在骨肉瘤组织中表达明显上调,其可以通过调节细胞周期中G1/S期的转换而影响骨肉瘤细胞生长。
Objective： To analyze the expression of microRNA-93 （miR-93） in osteosarcoma and to investigate the effect of miR-93 on the proliferation and cell cycle in osteosarcoma cell lines. Methods： TagMan MGB real-time PCR was used to detect the expression of miR-93 in 38 pairs of osteosarcoma and cartilaginous tissues. The over-expressed vector of miR-93 was constructed, and the miR-93 antisense nucleotide sequence was synthesized. Then they were transfected into osteosareoma cell lines U2-OS and MG-63. The cell proliferation was detected by MTF assay and cell cycle was detected by flow cytometry. Results： The 65.8% （25/38）of miR-93 was over expressed in osteosarcoma compared to cartilaginous tissues （P 〈 0.05）. The over expression of miR-93 could promote the growth of osteosarcoma cell line and delay the cell cycle in S phase. The reduction of miR-93 could inhibit the growth of osteosarcoma cell line and delay the cell cycle in G0/G1 phase. Conclusion： miR-93 was over expressed in osteosarcoma and could regulate the growth of osteosarcoma cells via mediating the transition of G1/S phase, which may become a target gene of osteosarcoma.
出处
《天津医药》
CAS
北大核心
2012年第12期1218-1221,共4页
Tianjin Medical Journal
作者简介
通讯作者:高志增,E-mail:690737188@qq.com
