邢晓静,谷小虎,马天飞.Biglycan及VEGF对结肠癌细胞增殖、凋亡能力的影响及分子机制[J].肿瘤学杂志,2014,20(6):471-476.
Biglycan及VEGF对结肠癌细胞增殖、凋亡能力的影响及分子机制
Effect of Biglycan and VEGF on Proliferation,Apoptosis of Colon Cancer Cells and Its Mechanism
投稿时间:2014-01-24  
DOI:10.11735/j.issn.1671-170X.2014.06.B008
中文关键词:  Biglycan  VEGF  结肠肿瘤  增殖  凋亡
英文关键词:Biglycan  VEGF  colon neoplasms  proliferation  apoptosis
基金项目:辽宁省科技攻关项目(2012225016);辽宁省自然科学基金(201102111);国家自然科学基金(81201968)
作者单位
邢晓静 辽宁省肿瘤医院 
谷小虎 辽宁省肿瘤医院 
马天飞 辽宁省肿瘤医院 
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中文摘要:
      摘 要:[目的] 观察Biglycan及VEGF对结肠癌细胞系HCT116增殖、凋亡的影响并探讨其可能机制。[方法]向转染Biglycan的细胞系HCT116中转染VEGF siRNA,设对照组(Mock)、空载和干扰对照组(Vector+siRNA-NC)、Biglycan cDNA和干扰对照组(Biglycan+siRNA-NC)及Biglycan cDNA和VEGF干扰组(Biglycan+siRNA-VEGF)。Western Blot检测Biglycan、VEGF及Ki67、PCNA、P21的表达;MTT检测细胞增殖情况;流式细胞术检测细胞凋亡及周期。[结果] 过表达Biglycan后,Ki67、PCNA和VEGF显著上调,P21显著下调。干扰VEGF后,上述3种周期蛋白的表达正好相反;Biglycan上调后,细胞增殖能力显著提高,下调VEGF后增殖能力又显著降低(P<0.05);Biglycan过表达后S期细胞总数(44.39%±1.80%)较Mock组(31.41%±1.81%)和Vector+siRNA-NC组(32.56%±1.07%)显著提高(P<0.01),凋亡率(0.12%±0.01%)较Mock组(1.75%±0.17%)和Vector+siRNA-NC组(1.83%±0.16%)显著下降(P<0.01);下调VEGF后S期细胞(20.76%±1.23%)显著降低(P<0.01),凋亡率(8.30%±0.71%)显著上升(P<0.01)。[结论] Biglycan通过促进VEGF的表达来促进结肠癌细胞的增殖并抑制其凋亡。
英文摘要:
      Abstract:[Purpose] To investigate the effect of Biglycan and VEGF on proliferation,apoptosis of HCT116 cells in vitro and its possible mechanisms. [Methods] VEGF siRNA was transfected into HCT116 cells which had stably transfected with Biglycan. Control group (Mock),empty plasmid and non-specific interference plasmid control group(Vector+siRNA-NC),Biglycan cDNA and non-specific interference plasmid group(Biglycan+siRNA-NC),Biglycan cDNA and VEGF siRNA cotransfection group(Biglycan+siRNA-VEGF) were set. The expressions of Biglycan,VEGF and Ki67,PCNA,P21 were detected by Western Blot. The proliferations of cells were detected by MTT. The apoptosis was determined by AV/PI staining and the cell cycle was determined by PI staining.[Results] Western Blot showed that compared with the control group,overexpression Biglycan significantly up-regulated Ki67,PCNA,VEGF and down-regulated p21(P<0.05). However,the expression of the three proliferation marker proteins were significantly opposited after reduction of VEGF(P<0.05). MTT showed that overexpression Biglycan significantly improved the proliferation ability and down-regulation of VEGF reduced this ability(P<0.01). Compared with Mock(31.41%±1.81%) and Vector+siRNA-NC(32.56%±1.07%),the number of cells in S phase(44.39%±1.80%) significantly increased after overexpression Biglycan(P<0.01) and decreased(20.76%±1.23%) after cotransfected with VEGF siRNA(P<0.01). Compared with Mock(1.75%±0.17%) and Vector+siRNA-NC (1.83%±0.16%),the apoptosis rate (0.12%±0.01%) significantly decreased after overexpression biglycan (P<0.01) and increased after cotransfected with VEGF siRNA(8.30%±0.71%)(P<0.01). [Conclusion] Biglycan promotes the proliferation and inhibit apoptosis of colon cancer cells by increasing the expression of VEGF.
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