摘要
目的研究靶向沉默增殖诱导配体(aproliferation-inducing ligand,APRIL)基因对人胃癌MGC803细胞增殖及细胞周期的影响。方法构建含有APRIL的siRNA序列的慢病毒载体并转染人胃癌MGC803细胞,以非特异性序列nonsilencing siRNA作为对照。通过Real-time PCR(RT-PCR)评价APRIL的沉默效率;MTT法检测沉默A-PRIL基因后,细胞增殖的变化;流式细胞仪检测细胞周期的变化。结果 RT-PCR测序证实,成功构建APRIL siR-NA慢病毒表达载体;与非特异性序列载体转染组相比,RT-PCR证实APRIL在慢病毒转染的细胞中表达显著降低(P<0.05);APRIL siRNA慢病毒载体转染后的MGC803细胞,细胞增殖显著降低,细胞周期被阻滞在G2/M期。结论靶向APRIL基因的慢病毒表达载体在体外能明显抑制人胃癌MGC803细胞的生长,为其在胃癌发病机制研究奠定了实验基础。
Objective To investigate the effects of a proliferation-inducing ligand（APRIL） gene silencing on cell proliferation and cell cycle of gastric cancer cell line（MGC803）.Methods Lentivirus containing siRNA sequence of APRIL was constructed,and infected the human gastric cancer MGC803 cells with nonspecific（NS） siRNA sequence as a control.APRIL mRNA expression was examined by real-time quantitative PCR（RT-PCR）;MTT assay was used to study the effects ofknockdown of APRIL in cell proliferation;cell cycle analysis was detected by flow cytometry.Results The results of sequencing demonstrated that the RNAi sequence targeting APRIL gene was successfully inserted into the lentiviral vector.Compared with non-specific sequence plasmid,the results of RT-PCR confirmed that expression of APRIL was much lower in APRIL-siRNA-transfected cells.（P 0.05）.Infected MGC803 cells with APRIL siRNA lentiviral vector,cell viability was significantly hindered,cell cycle arrested in G2/M phase.Conclusion Lentivirus containing siRNA targeting APRIL gene can inhibit the growth of human gastric cancer MGC803 cells in vitro and this lays a basis on the study of its pathogenesis in the gastric cancer.
出处
《中国实验诊断学》
2012年第12期2166-2169,共4页
Chinese Journal of Laboratory Diagnosis
基金
国家青年自然基金(30901702)
吉林省中青年领导人才创新团队项目(20111807)
吉林大学杰出青年基金(201005001)
关键词
增殖诱导配体
RNA干扰
胃癌
细胞增殖
细胞周期
a proliferation-inducing ligand
RNA interfering
gastric cancer
cell proliferation
cell cycle
