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Apoptosis of human colon carcinoma HT-29 cells induced by ceramide 被引量:4
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作者 Xiao-Feng Zhang Bai-Xiang Li +1 位作者 Chun-Yan Dong Rui Pen 《World Journal of Gastroenterology》 SCIE CAS CSCD 2006年第22期3581-3584,共4页
AIM: To investigate the effect of exogenous ceramideinduced apoptosis on human colon carcinoma HT-29 cells. METHODS: Light microscope, transmission electron microscope and fluorescence microscope were used to observ... AIM: To investigate the effect of exogenous ceramideinduced apoptosis on human colon carcinoma HT-29 cells. METHODS: Light microscope, transmission electron microscope and fluorescence microscope were used to observe the morphology change of apoptosis in HT-29 cells. Agarose gel electrophoresis was performed to detect the DNA fragment. Mitochondrial function was detected by MTT assay, mRNA expression of Bcl-2 family gene members was determined by reverse transcription polymerase chain reaction (RT-PCR) assay. RESULTS: After C2-ceramide treatment, typical characteristics of apoptosis, such as nuclear chromatin breakage, apoptotic body and DNA ladder, could be observed. After exposure to 50μmol/L C2-ceramide for 12 and 24 h, cell apoptosis was 64.1% and 81.3% respectively, which had a time-and dose-effect relationship. Mitochondrial function started to decrease from 6 h after exposure to ceramide. Meanwhile, ceramide up-regulated or down-regulated the mRNA expression of Bcl-2 family gene members. CONCLUSION: Ceramide induces apoptosis of human colon carcinoma HT-29 cells by affecting the expression of Bcl-2 family gene members and impacting the mitochondrial function. 展开更多
关键词 CERAMIDE APOPTOSIS human colon carcinomacells Bcl-2 family gene member Mitochondrial function
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神经酰胺诱导人结肠癌HT-29细胞凋亡的研究 被引量:7
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作者 张晓峰 李百祥 任锐 《卫生研究》 CAS CSCD 北大核心 2006年第5期537-540,共4页
目的探讨外源性神经酰胺诱导人结肠癌HT-29细胞的凋亡作用及对Bcl-2家族蛋白基因表达的影响。方法不同浓度C2-神经酰胺处理HT-29细胞,采用Hoechest荧光染色和琼脂糖凝胶电泳方法检测细胞凋亡,RT-PCR方法检测Bcl-2、Bcl-xl、Bax、Bad和Bi... 目的探讨外源性神经酰胺诱导人结肠癌HT-29细胞的凋亡作用及对Bcl-2家族蛋白基因表达的影响。方法不同浓度C2-神经酰胺处理HT-29细胞,采用Hoechest荧光染色和琼脂糖凝胶电泳方法检测细胞凋亡,RT-PCR方法检测Bcl-2、Bcl-xl、Bax、Bad和Bid基因表达水平。结果C2-神经酰胺诱导HT-29细胞发生核染色质断裂、出现凋亡小体等典型凋亡改变,50μmol/L神经酰胺作用12和24小时凋亡细胞率分别达到64·7%和81·3%。同时神经酰胺使Bax、Bad和Bid基因表达水平增高,使Bcl-2和Bcl-xl基因表达水平减弱,Bcl-2与Bax比值下降,呈现剂量-效应关系。结论外源性神经酰胺能通过上调或下调Bcl-2家族基因表达水平诱导人结肠癌HT-29细胞发生凋亡。 展开更多
关键词 神经酰胺 细胞凋亡 人结肠癌细胞 Bcl-2家族基因
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激活蛋白2家族基因真核表达载体的构建及其功能鉴定
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作者 米月 蒋琦炜 +2 位作者 沈雁婕 张德宇 叶棋浓 《基础医学与临床》 2022年第12期1829-1834,共6页
目的构建激活蛋白2(AP-2)家族真核表达载体,探究AP-2α、AP-2β、AP-2γ对人肝癌细胞系HepG2增殖及迁移的影响。方法模板为人卵巢文库,通过PCR扩增AP-2α、AP-2β以及AP-2γ,并与载体pCDNA3.0-FLAG连接得到真核表达载体pCDNA3.0-FLAG-A... 目的构建激活蛋白2(AP-2)家族真核表达载体,探究AP-2α、AP-2β、AP-2γ对人肝癌细胞系HepG2增殖及迁移的影响。方法模板为人卵巢文库,通过PCR扩增AP-2α、AP-2β以及AP-2γ,并与载体pCDNA3.0-FLAG连接得到真核表达载体pCDNA3.0-FLAG-AP-2α/β/γ;通过CCK8法、克隆形成实验和划痕实验分别验证AP-2α、AP-2β以及AP-2γ对肝癌HepG2细胞增殖和迁移的影响。结果成功构建了AP-2基因家族主要成员AP-2α、AP-2β、AP-2γ的真核表达载体,经酶切及基因测序鉴定重组质粒AP-2家族转录因子构建成功,并且能够成功表达;同时经过功能鉴定,显示AP-2家族蛋白可以显著抑制HepG2细胞的增殖以及迁移(P<0.05)。结论构建的AP-2家族基因真核表达载体转染至HepG2细胞后,明显抑制HepG2细胞增殖和迁移。 展开更多
关键词 ap-2基因家族 肝癌 真核表达 HEPG2细胞系
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Linkage analysis of a region on chromosome 2 with essential hypertension in Chinese families 被引量:2
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作者 朱鼎良 王厚义 +8 位作者 熊墨淼 初少莉 金力 王谷亮 何鑫 袁文涛 钱岳晟 赵光胜 黄薇 《Chinese Medical Journal》 SCIE CAS CSCD 2002年第5期654-657,146,共4页
OBJECTIVE: To verify the linkage of the candidate regions identified in a previous study (markers D2S168, D2S151, D2S142 on chromosome 2) with hypertension in Chinese families. METHODS: A genetic linkage study focused... OBJECTIVE: To verify the linkage of the candidate regions identified in a previous study (markers D2S168, D2S151, D2S142 on chromosome 2) with hypertension in Chinese families. METHODS: A genetic linkage study focused on chromosome 2 was performed on 240 Chinese families containing 856 patients with essential hypertension. A total of 1080 individuals were genotyped using 9 highly polymorphic microsatellite markers around the candidate regions on chromosome 2 with an average spacing of 5 cM. Non-parametric linkage (NPL), parametric linkage analysis and transmission-disequilibrium test (TDT) with the GENEHUNTER software were used to assess evidence for linkage. RESULTS: Linkage of a region on chromosome 2 around D2S151 and D2S142 with hypertension was confirmed by different statistical methods (NPL, LOD score and TDT). However, the linkage of D2S168 could not be replicated in this extension study. CONCLUSIONS: The data suggest that a region on chromosome 2 at or near the loci of D2S142 and D2S151 may harbor genes responsible for the development of essential hypertension in Chinese. 展开更多
关键词 Alleles China Chromosomes human Pair 2 family Health Female gene Frequency humans HYPERTENSION Linkage (genetics) Linkage Disequilibrium Male Microsatellite Repeats Research Support Non-U.S. Gov't
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