文章摘要
杨汉正,吴旭东,陈卫昌.PPARγ基因沉默抑制HCCLM3细胞侵袭与转移[J].南京医科大学学报,2008,28(7):889~893
PPARγ基因沉默抑制HCCLM3细胞侵袭与转移
PPARγ knockdown inhibited cell invasion and metastasis of HCCLM3 cells
投稿时间:2008-02-28  
DOI:10.7655
中文关键词: RNA干扰  PPARγ  HCCLM3  侵袭  转移
英文关键词: RNA interference  PPAR gamma  HCCLM3  invasion  metastasis
基金项目:
作者单位
杨汉正 盐城卫生职业技术学院免疫教研室,江苏 盐城 224006 
吴旭东 盐城市第一人民医院消化内科,江苏 盐城 224006 
陈卫昌 苏州大学附属第一医院消化内科,江苏 苏州 215006 
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中文摘要:
      目的:观察过氧化物酶体增殖物激活受体γ(PPARγ)基因沉默对HCCLM3细胞侵袭与转移的影响及可能机制?方法:应用Tanswell小室建立肿瘤细胞侵袭模型,PPARγ shRNA(pshPPARγ组)和pBi-hU6-DNA质粒(pshPPARγ-N组)转染HCCLM3细胞,并以未转染细胞为阴性对照(对照组),观察3组HCCLM3细胞黏附?迁移?侵袭和移动能力;RT-PCR法检测HCCLM3细胞基质金属蛋白酶(MMP)2?金属蛋白酶组织抑制剂(TIMP)2和整合素β1 mRNA表达?结果:pshPPARγ组PPARγ mRNA表达下调80.5%;pshPPARγ组跨过半透膜的细胞数为(68 ± 7)个,透过滤膜的细胞数为(17 ± 3)个,黏附的细胞数为(49 ± 9)个,细胞越过划痕的时间为(6.40 ± 1.14)天,与其他两组比较P均 < 0.01?pshPPARγ组MMP2?整合素β1表达下调,TIMP2表达上调?结论:PPARγ基因沉默能降低HCCLM3细胞的侵袭和转移能力;其可能经由整合素信号通路调节MMP2/TIMP2平衡而起作用?
英文摘要:
      Objective:To investigate the effect on invasion and metastasis of HCCLM3 cells blocked peroxisome proliferators-activated receptor gamma(PPARγ) expression by RNA interference method. Methods:Transwell chambers were used to construct tumor cell invasive models,PPARγ shRNA(pshPPARγ group) and plasmid pBi-hU6-DNA(pshPPARγ-N group) were transfected into HCCLM3 cells. Untreated HCCLM3 cells were regarded as negative control(control group). The changes of PPARγ expression in three groups were detected by semi-quantitive RT-PCR and Western blot analysis. Cell adhesion,migration,invasion and locomotion of cells were observed,Meanwhile,MMP2,TIMP2 and integrin β1 mRNA expression were detected by RT-PCR. Results:PPARγ expression in pshPPARγ group was 80.5% lower than the other two groups at mRNA level. In pshPPARγ groups,the number of cells crawled over semipermeable membrane of Tanswell chambers were 68 ± 7,the number of cells permeated through ECM were 17 ± 3,and the ability of cell adhesion decreased significantly. After 2 h,adhesion cell counts in pshPPARγ groups are 49 ± 9,the time when cells healed the wound were(6.40 ± 1.14)d. There were significant differences between pshPPARγ group and the other two groups(P < 0.01). In pshPPARγ group,MMP2 and integrin β1 mRNA of HCCLM3 cells decreased significantly,but TIMP2 increased. Conclusion:Knockdown PPARγ expression with RNA interference technology can significantly inhibit the invasion and metastasis of HCCLM3 cells, which might be the result of the decrease expression of integrin β1 to balance MMP2/TIMP2.
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