[1]周炜,姜政,张滨,等.短发夹RNA干扰胃癌低氧诱导因子-1α表达及其临床意义[J].第三军医大学学报,2009,31(02):152-155.
 ZHOU Wei,JIANG Zheng,ZHANG Bin,et al.RNA interference targeting hypoxia inducible factor-1α in BGC-823 cells inhibits cell proliferation and induces cell apoptosis[J].J Third Mil Med Univ,2009,31(02):152-155.
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短发夹RNA干扰胃癌低氧诱导因子-1α表达及其临床意义(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
31卷
期数:
2009年第02期
页码:
152-155
栏目:
论著
出版日期:
2009-01-30

文章信息/Info

Title:
RNA interference targeting hypoxia inducible factor-1α in BGC-823 cells inhibits cell proliferation and induces cell apoptosis
作者:
周炜姜政张滨向廷秀黄爱龙王丕龙
重庆医科大学:附属第一医院消化科,公共卫生学院社会医学教研室,病毒性肝炎研究所
Author(s):
ZHOU Wei JIANG Zheng ZHANG Bin XIANG Ting-xiu HUANG Ai-long WANG Pi-long
Department of Gastroenterology, First Affiliated Hospital, Department of Community Medicine, College of Public Health, Institute of Viral Hepatitis, Chongqing Medical University, Chongqing 400016, China
关键词:
胃癌RNA干扰低氧诱导因子-1α重组质粒细胞凋亡BGC-823细胞
Keywords:
gastric carcinoma RNA interference hypoxia inducible factor-1α recombinant plasmid cell apoptosis BGC-823 cell
分类号:
R394.3;R730.23;R735.2
文献标志码:
A
摘要:
目的    探讨RNA干扰对胃癌细胞低氧诱导因子-1α(HIF-1α)转录和蛋白表达的作用以及对胃癌细胞增殖和凋亡的影响。    方法    构建人HIF-1α基因的短发夹RNA(shRNA)真核表达载体并转染胃癌细胞BGC-823;采用RT-PCR和Western blot法检测HIF-1α mRNA、蛋白表达水平;MTT法检测转染后对胃癌细胞生长的抑制;TUNEL法观察细胞的凋亡情况;流式细胞术分析转染后胃癌细胞的细胞周期和凋亡率。    结果    成功构建了pshRNA-HIF-1α1和pshRNA-HIF-1α2重组质粒;转染BGC-823后,与空质粒组相比,pshRNA-HIF-1α1可有效抑制BGC-823细胞HIF-1α基因转录和蛋白质表达,抑制率分别可达93.4%和55.7%;MTT法显示转染后细胞生长明显受到抑制;流式细胞术结果表明转染后,其凋亡率明显高于空质粒组(P<0.05),BGC-823细胞增殖活性显著降低, G0~G1期细胞比例明显增加,S期与G2~M期细胞比例减少;TUNEL法显示转染后细胞核固缩,核染色质聚集或断裂;体内实验显示,pshRNA-HIF-1α1的抑瘤率为41.75%。    结论    体内、外初步实验证实pshRNA-HIF-1α1能有效抑制胃癌细胞BGC-823 HIF-1α基因表达,抑制胃癌细胞增殖,诱导其凋亡。
Abstract:
Objective    To study the expression of hypoxia inducible factor-1α (HIF-1α) in BGC-823 cells after RNA interference and the proliferation and apoptosis of BGC-823 cells.     Methods    The recombinant plasmid pshRNA-HIF-1α was constructed and transfected into gastric cancer cells BGC-823 by LipofectamineTM2000. Then the expressions of HIF-1α in BGC-823 cells were observed by RT-PCR and Western blotting, the cell proliferation was examined by MTT assay and the cell apoptosis by TUNEL and flow cytometry. The transfected cells were transplanted to nude mice to observe the inhibitory effect of transfection.     Results    The restriction enzyme digestion and sequencing result showed that the recombinant plasmid of pshRNA-HIF-1α had been constructed successfully. After it was transfected into BGC-823 cells, the genetic transcription and protein expression of HIF-1α were inhibited significantily (inhibitory ratio: 93.4% and 55.7%), compared with controls. The inhibitory ratio of cell proliferation in the pshRNA-HIF-1α transfected cells was higher than that of controls, and the cell ratio was increased in G0/G1phase, and decreased in S and G2/M phases. The apoptotic ratio of the cells transfected with pshRNA-HIF-1α was also higher than that of controls. Karyopycnosis, nuclear chromosomal condensation and segmentation were observed by TUNEL. In vivo, pshRNA-HIF-1α1 was proved to be able to inhibit the proliferation and induce the apoptosis of BGC-823 cells by restraining the expressions of HIF-1α, whose tumor inhibition rate was 41.75%.     Conclusion    Our results show that the recombinant plasmid pshRNA-HIF-1α can inhibit the genetic transcription and protein expression of HIF-1α, and suppress the proliferation and induce the apoptosis of gastric cancer cells BGC-823 in vivo and in vitro.

参考文献/References:

周炜, 姜政, 张滨, 等. 短发夹RNA干扰胃癌低氧诱导因子-1α表达及其临床意义[J]. 第三军医大学学报, 2009, 31(2):152-155.

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更新日期/Last Update: 2009-01-08