Growth inhibition and apoptosis induction effects of pLXSN-Tum-5 in human umbilical vein endothelial cells
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摘要: 目的 探讨pLXSN-Tum-5病毒颗粒对人脐静脉内皮细胞(HUVEC)的生长抑制和诱导凋亡作用及机制。方法 采用不同浓度的pLXSN-Tum-5病毒颗粒作用HUVEC,利用噻唑蓝法和Hoechst-33342染色法检测细胞生长及其形态变化;RT-PCR和Western blotting方法检测细胞中Bax、Bcl-2、caspase-3 mRNA和蛋白表达变化。结果 与对照组(pLXSN)比较,pLXSN-Tum-5病毒颗粒转染组HUVEC的存活率明显降低,呈剂量效应关系;细胞内可见凋亡小体;对照组HUVEC内Bcl-2和caspase-3 mRNA和蛋白表达分别为(0.721±0.041)、(0.654±0.034)和(0.956±0.032)、(0.356±0.054);与对照组比较,20 μmol/L pLXSN-Tum-5病毒颗粒转染组HUVEC内Bcl-2和caspase-3 mRNA和蛋白水平[分别为(1.134±0.0524)、(1.012±0.0641)和(1.612±0.067)、(0.712±0.0647)]明显上调(P<0.05)。结论 Tum-5可抑制HUVEC增殖,诱导HUVEC凋亡,其机制可能与上调Bax/Bcl-2和caspase-3表达有关。
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关键词:
- 肿瘤抑素(tumstatin) /
- Tum-5 /
- 细胞凋亡 /
- 人脐静脉内皮细胞(HUVEC) /
- 抗血管生成
Abstract: Objective To explore growth inhibition and apoptosis induction effects of pLXSN-Tum-5 in human umbilical vein endothelial cells (HUVEC) and the mechanism of the effects.Methods HUVEC were treated with pLXSN-Tum-5 virus particles at different concentrations.Hoechst-33342 staining and 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide (MTT) assay were utilized to monitor the growth states and morphological changes of HUVEC.Changes in the expressions of intracellular B cell lymphoma 2 (Bcl-2),Bcl-2-associated X (Bax),and caspase-3 at mRNA and protein levels were determined with real-time reverse transcription PCR (RT-PCR) and Western blot.Results The survival rate of HUVEC transfected with pLXSN-Tum-5 virus particles was significantly decreased in a dose-effect manner compared to that of the control (with pLXSN).Apoptotic bodies were observed in the HUVEC transfected with pLXSN-Tum-5.Significantly upregulated mRNA and protein expressions of Bcl-2 (1.134±0.0524 vs.0.721±0.041 and 1.012±0.0641 vs.0.654±0.034) and caspase-3 (1.612±0.067 vs.0.956±0.032 and 0.712±0.0647 vs.0.356±0.054) were observed in the HUVEC transfected with 20 mol/L pLXSN-Tum-5 virus particles compared to those in the control HUVEC (P<0.05 for all).Conclusion Human Tum-5 could inhibit proliferation and induce apoptosis in HUVEC,and the mechanism of the effects may be related to upregulated expressions of Bax/Bcl-2 and caspase-3 in the HUVEC.-
Key words:
- tumstatin /
- Tum-5 /
- cell apoptosis /
- human umbilical vein endothelial cell /
- anti-angiogenesis
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[1] Kizaka-Kondoh S,Kuchimaru T,Kadonosono T.Pathophysiological response to hypoxia-from the molecular mechanisms of malady to drug discovery:hypoxia-inducible factor-1(HIF-1)-active cells as a target for cancer therapy[J].Pharmacol Sci,2011,115(4):440-445. [2] Sudhakar A,Boosani CS.Inhibition of tumor angiogenesis by tumstatin:insights into signaling mechanisms and implications in cancer regression[J].Pharm Res,2008,25(12):2731-2739. [3] Floquet N,Pasco S,Ramont L,et al.The antitumor properties of the alpha3(Ⅳ)-(185-203) peptide from the NC1 domain of type Ⅳ collagen (tumstatin) are conformation-dependent[J].J Biol Chem,2004,279(3):2091-2100. [4] Hamano Y,Zeisberg M,Sugimoto H,et al.Physiological levels of tumstatin,a fragment of collagen IV 3 chain,are generated by MMP-9 proteolysis and suppress angiogenesis via V3 integrin[J].Cancer Cell,2003,3(6):589-601. [5] You Y,Xue X,Li M,et al.Inhibition effect of pcDNA-tum-5 on the growth of S180 tumor[J].Cytotechnology,2008,56(2):97-104. [6] 盖晓东,罗宏,历春,等.tum-5基因逆转录病毒载体及包装细胞株的构建[J].中国老年学杂志,2009,5(29):1194-1196. [7] Folkman J.Tum of angiogenesis therapeutic implications[J].Engl NJ Med,1971,285(21):1182-1186. [8] Carmeliet P,Jain RK.Angiogensis in cancer and other diseases[J].Nature,2000,407(6801):249-257. [9] Folkman J,Kalluri R.Cancer without disease[J].Nature,2004,427(6977):787. [10] Naumov GN,Folkman J,Straume O.Tumor dormancy due to failure of angiogenesis[J].Clin Ex PMetastasis,2009,26(1):51-60. [11] Maeshima Y,Colorado PC,Torre A,et al.Distinct antitumor properties of a type Ⅳ collagen domain derived from basement membrane[J].J Biol Chem,2000,275(28):21340-21348.
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