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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 & AIM: Neurotrophic factors such as nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), and neurotrophic factor 3/4 (NT) have been shown to be synthesized in various cells both inside and outside the nervous system -3/4). These neurotrophic factors can not only promote the survival, proliferation and apoptosis of nerve cells, but also the activity of cancer cells invade nerve tissue. However, whether BDNF correlates with the biological behavior of lung cancer cells has not been reported yet. This article aims to investigate the effect of BDNF on hydrogen peroxide (H2O2) -induced apoptosis in human lung cancer cell line YTMLC-90. Methods: Human lung cancer cell line YTMLC-90 was transfected with pSVCEPBFCAT, a promoter and enhancer element of human COLI1 gene and BDNF cDNA at its 3 ’terminus. The eukaryotic expression of BDNF was induced by tetrakis Cell proliferation was detected by methylthiazolyl tetrazolium (MTT) colorimetric assay. Fluorescence microscopy and electron microscopy were used to observe the changes of cell morphology and ultrastructure. Chromatin DNA fragmentation was detected by agarose gel electrophoresis. Results: The growth inhibition rate of YTMLC-90 cells transfected with pSVCEPBFCAT was 30.0% after treated with 200μmol / LH2O2 for 24 h and 84.60% for YTMLC-90 cells transfected with empty vector pSVCEPCAT and without YTMLC-90 80.00%, there was significant difference between the experimental group and the two control groups (P <0.01). The control group showed morphological and biochemical changes of apoptosis, such as cytoplasm and nuclear pyknosis, Chromatin breakage and DNA electrophoresis showed a ladder-like pattern. However, no apoptosis was observed in YTMLC-90 cells transfected with pSVCEPBFCAT. Conclusion: BDNF promotes YTMLC-90?