目的研究重组腺相关病毒介导的人内皮抑素对裸鼠鼻咽癌肝脏异位移植瘤生长和转移的抑制作用。方法检测携带人内皮抑素的重组腺相关病毒(RECOMBINANT ADENO-ASSOCIATED VIRUSCARRYING HUMAN ENDOSTATIN GENE,RAAV-HENDO)体外感染效率、...目的研究重组腺相关病毒介导的人内皮抑素对裸鼠鼻咽癌肝脏异位移植瘤生长和转移的抑制作用。方法检测携带人内皮抑素的重组腺相关病毒(RECOMBINANT ADENO-ASSOCIATED VIRUSCARRYING HUMAN ENDOSTATIN GENE,RAAV-HENDO)体外感染效率、蛋白表达及生物活性。建立裸鼠鼻咽癌肝脏异位移植瘤动物模型,经尾静脉分别注射携带人内皮抑素基因的RAAV-HENDO(HENDO组)、携带增强型绿色荧光蛋白基因的RAAV-EGFP(RAAV-CARRYING ENHANCED GREEN FLUORESCENT PROTEIN GENE,RAAV-EGFP,EGFP组)和磷酸缓冲液(PBS组),观察动物肝脏成瘤、肺转移及生存期等情况,免疫组化检测肿瘤微血管密度,原位末端标记(TERMINAL DEOXYNUCLEOTIDYL TRANSFERASE-MEDIATED DUTP NICK-END-LABELING,TUNEL)法检测肿瘤细胞凋亡指数。结果体外实验表明:RAAV体外感染效率达98%;免疫荧光染色显示内皮抑素蛋白主要表达于细胞质;RAAV-HENDO感染细胞培养上清对ECV304细胞72H增殖抑制率为67·3%。动物实验表明:相对PBS组,HENDO组抑瘤率为70·7%;HENDO组、EGFP组、PBS组肺转移率分别为0·0%、50·0%、66·7%;微血管平均(X-±S,以下同)密度分别为(3·67±1·63)、(19·67±2·16)、(22·50±3·02),组间比较差异有统计学意义(P<0·01);凋亡平均指数分别为(28·83±5·27)%、(6·17±2·79)%、(4·50±2·17)%,组间比较差异有统计学意义(P<0·01);3组动物平均生存期分别为(36·50±8·46)D、(24·00±5·66)D、(21·17±3·92)D,组间比较差异有统计学意义(P<0·01)。结论重组腺相关病毒介导的内皮抑素能有效抑制裸鼠鼻咽癌肝脏异位移植瘤的生长和转移,其可能机制为抑制肿瘤新生血管形成,诱导肿瘤细胞凋亡。展开更多
BACKGROUND： Inhibitory signals, i.e. neurite growth inhibitors （NGIs）, presenting on central nervous system （CNS） myelin have been shown to play a crucial role in inhibiting lesioned axonal sprouting and leading ...BACKGROUND： Inhibitory signals, i.e. neurite growth inhibitors （NGIs）, presenting on central nervous system （CNS） myelin have been shown to play a crucial role in inhibiting lesioned axonal sprouting and leading to less functional recovery. Vaccines targeting NGIs may provide multifactorial protection against brain insults by overcoming the inhibitory effects of these NGIs and boosting the body＇s immune repair mechanisms. OBJECTIVE： To evaluate the effect of poststroke DNA immunization against NGIs on the rehabilitation for sensorimotor function of rat models of local cerebral ischemia. DESIGN： Completely randomized grouping design, and controlled experiment. SETTING： Brain Injury Research Laboratory, Department of Neurosurgery, National Neuroscience Institute, Singapore. MATERIALS： Sixty adult male Sprague-Dawley rats ranging in age from 45 to 120 days and in weight from 180 to 250 grams were provided by Animal Center of Department of Anatomy, Faculty of Medicine, National University of Singapore. pcDNA3.1（＋）-neurite growth inhibitors （pcDNA-NGIs） a gift was provided by Dr. Xiao from Department of Clinical Research, Singapore General Hospital, Singapore. METHODS： The experiment was carried out at Brain Injury Research Laboratory, Department of Neurosurgery, National Neuroscience Institute, Singapore from August 2003 to April 2005. （1）The involved rats were randomized into 3 groups： pcDNA-NGIs group （group A）, pcDNA3.1 （＋） group （group B） and model group （group C）, with 20 rats in each group. Left focal cerebral ischemia （FCI） was permanently induced through middle cerebral artery occlusion （MCAO） with the assistance of an operating microscope. Successful MCAO was determined by a 20% decrease to baseline in the ipsilateral cerebral blood flow. 100 μg of pcDNA-NGIs eluted in phosphate-buffered saline （PBS） was intramuscularly injected into the tibial muscle once a week after MCAO for 6 weeks in group A. As control, pcDNA3.1 （＋） was also adm展开更多
文摘目的研究重组腺相关病毒介导的人内皮抑素对裸鼠鼻咽癌肝脏异位移植瘤生长和转移的抑制作用。方法检测携带人内皮抑素的重组腺相关病毒(RECOMBINANT ADENO-ASSOCIATED VIRUSCARRYING HUMAN ENDOSTATIN GENE,RAAV-HENDO)体外感染效率、蛋白表达及生物活性。建立裸鼠鼻咽癌肝脏异位移植瘤动物模型,经尾静脉分别注射携带人内皮抑素基因的RAAV-HENDO(HENDO组)、携带增强型绿色荧光蛋白基因的RAAV-EGFP(RAAV-CARRYING ENHANCED GREEN FLUORESCENT PROTEIN GENE,RAAV-EGFP,EGFP组)和磷酸缓冲液(PBS组),观察动物肝脏成瘤、肺转移及生存期等情况,免疫组化检测肿瘤微血管密度,原位末端标记(TERMINAL DEOXYNUCLEOTIDYL TRANSFERASE-MEDIATED DUTP NICK-END-LABELING,TUNEL)法检测肿瘤细胞凋亡指数。结果体外实验表明:RAAV体外感染效率达98%;免疫荧光染色显示内皮抑素蛋白主要表达于细胞质;RAAV-HENDO感染细胞培养上清对ECV304细胞72H增殖抑制率为67·3%。动物实验表明:相对PBS组,HENDO组抑瘤率为70·7%;HENDO组、EGFP组、PBS组肺转移率分别为0·0%、50·0%、66·7%;微血管平均(X-±S,以下同)密度分别为(3·67±1·63)、(19·67±2·16)、(22·50±3·02),组间比较差异有统计学意义(P<0·01);凋亡平均指数分别为(28·83±5·27)%、(6·17±2·79)%、(4·50±2·17)%,组间比较差异有统计学意义(P<0·01);3组动物平均生存期分别为(36·50±8·46)D、(24·00±5·66)D、(21·17±3·92)D,组间比较差异有统计学意义(P<0·01)。结论重组腺相关病毒介导的内皮抑素能有效抑制裸鼠鼻咽癌肝脏异位移植瘤的生长和转移,其可能机制为抑制肿瘤新生血管形成,诱导肿瘤细胞凋亡。
基金the grants from the National Medical Research Council and Health Research of Singapore
文摘BACKGROUND： Inhibitory signals, i.e. neurite growth inhibitors （NGIs）, presenting on central nervous system （CNS） myelin have been shown to play a crucial role in inhibiting lesioned axonal sprouting and leading to less functional recovery. Vaccines targeting NGIs may provide multifactorial protection against brain insults by overcoming the inhibitory effects of these NGIs and boosting the body＇s immune repair mechanisms. OBJECTIVE： To evaluate the effect of poststroke DNA immunization against NGIs on the rehabilitation for sensorimotor function of rat models of local cerebral ischemia. DESIGN： Completely randomized grouping design, and controlled experiment. SETTING： Brain Injury Research Laboratory, Department of Neurosurgery, National Neuroscience Institute, Singapore. MATERIALS： Sixty adult male Sprague-Dawley rats ranging in age from 45 to 120 days and in weight from 180 to 250 grams were provided by Animal Center of Department of Anatomy, Faculty of Medicine, National University of Singapore. pcDNA3.1（＋）-neurite growth inhibitors （pcDNA-NGIs） a gift was provided by Dr. Xiao from Department of Clinical Research, Singapore General Hospital, Singapore. METHODS： The experiment was carried out at Brain Injury Research Laboratory, Department of Neurosurgery, National Neuroscience Institute, Singapore from August 2003 to April 2005. （1）The involved rats were randomized into 3 groups： pcDNA-NGIs group （group A）, pcDNA3.1 （＋） group （group B） and model group （group C）, with 20 rats in each group. Left focal cerebral ischemia （FCI） was permanently induced through middle cerebral artery occlusion （MCAO） with the assistance of an operating microscope. Successful MCAO was determined by a 20% decrease to baseline in the ipsilateral cerebral blood flow. 100 μg of pcDNA-NGIs eluted in phosphate-buffered saline （PBS） was intramuscularly injected into the tibial muscle once a week after MCAO for 6 weeks in group A. As control, pcDNA3.1 （＋） was also adm
