RNA interference targeting hypoxia inducible factor-1α in BGC-823 cells inhibits cell proliferation and induces cell apoptosis

周炜; 姜政; 张滨; 向廷秀; 黄爱龙; 王丕龙

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RNA interference targeting hypoxia inducible factor-1α in BGC-823 cells inhibits cell proliferation and induces cell apoptosis

更新时间：2026-01-09

- RNA interference targeting hypoxia inducible factor-1α in BGC-823 cells inhibits cell proliferation and induces cell apoptosis
- Journal of Army Medical University Vol. 31, Issue 2, Pages: 152-155(2009)
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  CLC： R394.3;R730.23;R735.2
- Published：2009
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周炜,姜政,张滨,向廷秀,黄爱龙,王丕龙.短发夹RNA干扰胃癌低氧诱导因子-1α表达及其临床意义[J].陆军军医大学学报(原第三军医大学学报),2009,31(02):152-155.

　ZHOU Wei,JIANG Zheng,ZHANG Bin,XIANG Ting-xiu,HUANG Ai-long,WANG Pi-long.RNA interference targeting hypoxia inducible factor-1α in BGC-823 cells inhibits cell proliferation and induces cell apoptosis[J].J Amry Med Univ (J Third Mil Med Univ),2009,31(02):152-155.
周炜,姜政,张滨,向廷秀,黄爱龙,王丕龙.短发夹RNA干扰胃癌低氧诱导因子-1α表达及其临床意义[J].陆军军医大学学报(原第三军医大学学报),2009,31(02):152-155. DOI：

　ZHOU Wei,JIANG Zheng,ZHANG Bin,XIANG Ting-xiu,HUANG Ai-long,WANG Pi-long.RNA interference targeting hypoxia inducible factor-1α in BGC-823 cells inhibits cell proliferation and induces cell apoptosis[J].J Amry Med Univ (J Third Mil Med Univ),2009,31(02):152-155. DOI：

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（P0.05），BGC-823细胞增殖活性显著降低

～G

期细胞比例明显增加，S期与G

～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 Lipofectamine

2000. 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 G

phase

and decreased in S and G

/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.

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