摘要
背景与目的:已证实在神经系统内外多种细胞中均可合成神经营养因子,如神经生长因子(nervegrowthfactor,NGF)、脑源性神经营养因子(brainderivedneurotrophicfactor,BDNF)和神经营养素3/4(NT-3/4)。这些神经营养因子不仅能促进神经细胞的存活、增殖和凋亡,也与癌细胞侵犯神经组织的活性有关;但BDNF与肺癌细胞的生物学行为是否相关尚未见报道。本文旨在研究BDNF对过氧化氢(H2O2)诱导人肺癌细胞YTMLC-90凋亡的作用。方法:用含人I型胶原a1链(COLIA1)基因启动子和增强子元件及在其3'端接BDNFcDNA的微基因pSVCEPBFCAT,转染人肺癌细胞YTMLC-90并驱动BDNF异位表达,采用四甲基偶氮唑蓝(MTT)显色法检测细胞增殖,吖啶橙荧光染色显微镜和电镜观察细胞形态及其超微结构的改变,琼脂糖凝胶电泳检测染色质DNA断裂。结果:经200μmol/LH2O2作用24h,转染pSVCEPBFCAT的YTMLC-90细胞生长抑制率为30.0%,而未转染YTMLC-90和转染空载体pSVCEPCAT的YTMLC-90细胞生长抑制率分别为84.60%和80.00%,实验组与两个对照组之间的差异均有显著性(P<0.01);对照组YTMLC-90细胞可见凋亡特有的形态学及生物化学改变,如胞浆和核固缩、染色质断裂、DNA电泳呈梯状条带,而转染pSVCEPBFCAT的YTMLC-90细胞未发现上述凋亡特征。结论:BDNF促进YTMLC-90?
BACKGROUND &OBJECTIVE:It has been shown that neurotrophic factors such as nerve g rowth factor (NGF ),brain derived neurotrophic factor (BDNF),and neurotrophin(NT-3/4)are synthesized in a variety of cells inside and outside t he nervous system.These factors are not only able to promote neural survival,pro liferation and apoptosis of neural c ells but also relevant to the activity of neur al invasion of tumors.It has not been reported to date whether BDNF may play roles in the biolog ical behavior of human lung cancer cells.The aim of this experim ent was to investig ate the effect of B DNF on hydrog en peroxide (H2O2)-induced apoptosis in the human lung cancer cell line YTMLC-90.METHODS :The minig ene pSVCEPBFCAT containin g the promoter and enhancer elements of the human a1(I)collag en g ene(COLIA1)at its 3' terminus followed by hBDNF g ene cDNA was transfected and derived BDNF ectopic expression in the human lung cancer cells.The cell proliferatio n was measured by MTT assay.The morpholog ical and ultra-structural chang es o f apoptotic cells were observed by mic roscopy with fluorescent stain of ac ridine orang e and electron microscopy.The DNA frag mentation was examined by ag arose g el electrophoresis.RESULTS :After exposure of g rowing cells to 200μmol /L H 2 O 2 for 24hours ,the inhibition rate of c ell g rowth was 30%in the pSVCEPBFCAT-transfected YTMLC-90,84.60%in controls of non-transfected YTMLC-90,and 80.00%in pSVCEPCAT-transfected YTMLC-90,respectively(P<0.001).The morpholog ical and biochemical chang es of apoptotic cells such as shrinkag e of cytoplasm and nu cleus ,frag mentation of the chromatin,and ladder pattern of DNA were common ly observed in the cell population of controls ,but these apoptotic featu res were not discovered in the pSVCEPBFCAT-transfected YTMLC-90.CONCLUSION:BDNF markedly inhibits H 2 O 2 cytotoxicity on human lung cancer cell YTMLC-90by promoting YTMLC-90proliferation and antag onizing H 2 O 2 -induced apoptosis.
出处
《癌症》
SCIE
CAS
CSCD
北大核心
2003年第9期938-942,共5页
Chinese Journal of Cancer
