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。
[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 siR- NA 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 MTF. 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 ceils 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.
出处
《肿瘤学杂志》
CAS
2014年第6期471-476,共6页
Journal of Chinese Oncology
基金
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)
