miRNA-214对胃癌细胞BGC823|MKN45的细胞周期和凋亡的影响
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郑长黎 E-mail:changlizheng1125@yahoo.com.cn

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The effect of miRNA-214 on cell cycle and apoptosis of gastric cancer cell lines BGC823 and MKN45
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    摘要:

    目的:探讨miRNA-214对胃癌细胞的细胞周期和凋亡的影响,及其对靶基因PTEN蛋白表达的影响。
    方法:荧光定量PCR法检测人低分化胃癌细胞BGC823、MKN45和人正常胃黏膜细胞GES-1中miRNA-214的表达;检测瞬时转染反义核苷酸(miRNA-214 抑制剂)后的miRNA-214的表达;流式细胞术分析转染后的细胞生长周期和凋亡率的变化;免疫荧光检测miRNA-214抑制剂对靶基因PTEN表达的影响。
    结果:miRNA-214 在人胃癌细胞BGC823及MKN45中表达上调(P<0.05)。转染miRNA-214 抑制剂后能使BGC823中miRNA-214的表达下调(P<0.05),且BGC823和MKN45在转染组的G1期细胞较未转染组明显增高[分别为(60.20±3.38)% vs. (49.33±7.99)% (P<0.05);(69.90±0.28)% vs. (54.85±0.64)% (P<0.05)],S期细胞在转染组较未转染组明显降低[(21.87±3.20)%,(18.25±1.34)%,P<0.05)],但对细胞凋亡率无影响(P>0.1)。免疫荧光显示,转染miRNA-214抑制剂后PTEN呈强表达。
    结论:(1)人胃癌细胞系BGC823,MKN45中的miRNA-214呈高表达;(2)miRNA-214可使这两株细胞的G1期细胞减少,S期细胞增多,下调PTEN可能是其作用机制之一;(3)miRNA-214对上述两株细胞的凋亡无影响。

    Abstract:

    Objective:To identify the effects of miRNA-214 on cell cycle and apoptosis of gastric cancer cells, and its influence on PTEN protein level.
    Methods:miRNA-214 in poorly differentiated human gastric cancer cell line BGC823, MKN45 and normal gastric mucosal cell line GES-1 was detected by real-time PCR. Antisense-miRNA-214 oligonucleotides(miRNA-214 inhibitor) were  transfected transiently into gastric cancer cell lines to down-regulate the expression of miRNA-214. The cell cycle and apoptosis rate were studied by flow cytometry. PTEN, the target gene of miRNA-214, was detected by immunofluorescence.
    Results:The miRNA-214 was upregulated in gastric cancer cell BGC823 and MKN45 (P<0.05) compared with normal gastric mucosal cell line GES-1. The transfection of miRNA-214 inhibitor downregulated miRNA-214 expression in BGC823 (P<0.05). G1-phase cells were increased in BGC823 and MKN45, [(60.20±3.38)% vs. (49.33±7.99)% (P<0.05);(69.90±0.28) vs. (54.85±0.64)% (P<0.05) respectively] compared with the control groups. But the alteration of apoptotic rate was not significant(P>0.1). The immunofluorescence result showed that PTEN was upregulated after transfection in both cell lines.
    Conclusions:(1)miRNA-214 is upregulated in human gastric cancer cell BGC823 and MKN45. (2)miRNA-214 could reduce the G1-phase cells and increase S-phase cells in BGC823 and MKN45, and the downregulation of PTEN may be one of the mechanisms. (3)miRNA-214 has no effect on apoptosis of BGC823 and MKN45.

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熊欣|郑长黎. miRNA-214对胃癌细胞BGC823|MKN45的细胞周期和凋亡的影响[J].中国普通外科杂志,2010,19(12):1311-1315.
DOI:10.7659/j. issn.1005-6947.2010.12.016

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  • 收稿日期:2010-03-22
  • 最后修改日期:2010-09-20
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  • 在线发布日期: 2010-12-15