摘要
目的检测microRNA-365(miR-365)对SOSP-9607骨肉瘤细胞增殖和凋亡的影响。方法利用人工合成的miR-365模拟物以及miR-365抑制物,瞬时转染SOSP-9607骨肉瘤细胞。实时定量PCR(qRT-PCR)检测细胞中miR-365的水平;流式细胞术检测SOSP-9607骨肉瘤细胞的细胞周期与凋亡;MTT法检测细胞增殖能力的变化;qRT-PCR检测细胞中KRAS的水平。结果 miR-365模拟物上调骨肉瘤细胞miR-365表达水平,抑制细胞增殖,细胞G1期增加、S/G2期减少,同时凋亡增加,KRAS基因的表达下降;miR-365抑制物下调SOSP-9607骨肉瘤细胞miR-365的表达水平,促进骨肉瘤细胞增殖,同时抑制凋亡,上调KRAS基因的表达。结论 miR-365抑制SOSP9607骨肉瘤细胞增殖并促进其凋亡,可能通过调节KRAS来发挥抑癌作用。
Objective To investigate the effect of miR-365 on the proliferation and apoptosis in SOSP-9607 osteosarcoma cells. Methods SOSP-9607 cells were transiently transfected with miR-365 mimic or miR-365 inhibitor which were artificially synthesized. The expression of miR-365 was detected by real-time PCR（ qRT-PCR）; the cell cycle and apoptosis was analyzed by flow cytometry; the cell proliferation was observed by MTT assay; and the level of KRAS was determined by qRT-PCR and Western blotting. Results miR-365 mimic up-regulated the expression level of miR-365 in SOSP-9607 osteosarcoma cells. After miR-365 mimic transfection,the proliferation of SOSP-9607 cells was inhibited; the cell cycle was arrested in G1 phase; the apoptosis rate increased and the expression of KRAS was reduced in miR-365 mimic transfected cells. On the contrary,when miR-365 inhibitor was transfected into SOSP-9607 cells,the expression level of miR-365 was significantly reduced along with promoted cell proliferation,suppressed cell apoptosis and increased KRAS expression.Conclusion miR-365 could inhibit the proliferation and promote the apoptosis in SOSP-9607 osteosarcoma cells probably by mediating the expression of KRAS.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2016年第1期44-48,共5页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金(30901784
81272072)
作者简介
高金鉴（1986-），男，山西太原人，硕士研究生Tel：18192509639；E-mail：mrgaojinjian@163．com
通讯作者，王涛，E-mail：fmmuwangt@fmmu．edu．cn；
单乐群，E-mail：drshanlq@fmmu．edu．cn
