Ghrelin抑制结肠上皮细胞凋亡及其信号通路
发布时间：2021-01-17 19:11
目的研究ghrelin是否能够抑制结肠上皮细胞凋亡,观察一些凋亡相关蛋白表达水平的变化,并进一步探讨ghrelin是否通过PI3K/Akt和ERK1/2信号通路抑制结肠上皮细胞凋亡。方法1.通过RT-PCR检测HT-29结肠癌细胞株上是否有GHS-R1a的表达。2.将细胞分为对照组和处理组,处理组细胞由5-Fu（80μM）单独或与ghrelin(10-9M-10-6M)共同处理24h-48h。3.通过MTT法对各组细胞进行生长活性的检测,计算细胞生存率。4.通过DAPI染色后在荧光显微镜下观察各组细胞核的形态。5.通过Annexin V/PI双染色后行流式细胞术,测定并比较各组细胞的凋亡率。6.通过Western-blot观察凋亡相关蛋白caspase3、bcl-2、bax及信号传导蛋白Akt、P-Akt、ERK1/2、P-ERK1/2的表达水平在各组中的变化。结果1.RT-PCR检测到HT-29细胞上有GHS-R1a的表达。2.MTT法显示ghrelin呈剂量依赖性地降低了5-Fu引起的HT-29细胞生长抑制,提高了细胞生存率（P＜0.05...
【文章来源】：复旦大学上海市 211工程院校 985工程院校 教育部直属院校
【文章页数】：52 页
【学位级别】：硕士
【部分图文】：
凋亡调节通路
选用卜aetion，引物序列为(「):5一e丁ee奸eeAee仃eAee6仃ee一3，引物扩增长度为26sbp。扩增以94oC305，50℃305，72oCZmin为一循环，10min。图1为PcR产物的凝胶电泳扫描图像，第道是内参p一action，其位置在250bp偏上;第三oobp之间有一明显条带，与GHs一Rla大小相符有GHS一Rla的表达。
图2ghrelin对H下29细胞生长的影响注:与对照组比较，*p<0.05;与s一「u组比较，△P<0，05。OAPI染色典型的凋亡细胞核呈现染色质的浓缩、边集、核固缩、核碎裂，染色3中A、B、e依次是对照组、5一「uso协M处理组，s一「u+Gio一6M处理组，时间为24h。A图中细胞核基本都是正常形态;B图中可见核固缩、
【参考文献】：
期刊论文
[1]Different effects of ghrelin, des-acyl ghrelin and obestatin on gastroduodenal motility in conscious rats[J]. Mineko Fujimiya,Akihiro Asakawa,Koji Ataka,Ikuo Kato,Akio Inui. World Journal of Gastroenterology. 2008(41)
[2]Ghrelin and gastric acid secretion[J]. Koji Yakabi,Junichi Kawashima,Shingo Kato. World Journal of Gastroenterology. 2008(41)
[3]T-2 toxin-induced apoptosis involving Fas,p53,Bcl-xL,Bcl-2,Bax and caspase-3 signaling pathways in human chondrocytes[J]. Jing-hong CHEN Jun-ling CAO,Yong-lie CHU,Zhi-lun WANG,Zhan-tian YANG,Hong-lin WANG (Ministry of Education Key Laboratory of Environment and Genes related to Diseases,Institute of Endemic Diseases,School of Medicine,Xi’an Jiaotong University,Xi’an 710061,China). Journal of Zhejiang University（Science B:An International Biomedicine & Biotechnology Journal）. 2008(06)
[4]MAPKs在Bcl-2家族调控细胞凋亡中的作用[J]. 古晓娜,刘占旗,战景明,谢满廷. 国外医学(卫生学分册). 2008(02)
[5]PI3K-Akt信号通路与肿瘤细胞凋亡关系的研究进展[J]. 黄秀兰,崔国辉,周克元. 癌症. 2008(03)
[6]Oridonin induces apoptosis via PI3K/Akt pathway in cervical carcinoma HeLa cell line[J]. Hong-zhen HU~2,Yue-bo YANG~3,Xiang-dong XU~4,Hong-wei SHEN~5,Yi-min SHU~6,Zi REN~7,Xiao-mao LI~3,Hui-ming SHEN~3,Hai-tao ZENG~(3,8)~2 Department of Gynecology and Obstetrics,Yijishan Hospital,Wannan Medical College,Wuhu 241001,China;~3 Department of Gynecologyand Obstetrics,the Third Affiliated Hospital,Sun Yat-sen University,Guangzhou 510080,China;~4 Department of Surgery,the First AffiliatedHospital,Sun Yat-sen University,Guangzhou 510080,China;~2 Department of Gynecology and Obstetrics,the First Affiliated Hospital,SunYat-sen University,Guangzhou 510080.China;~6 Department of Obstetrics and Gynecology,IVF Program,Stanford University MedicalCenter,Palo Alto,CA 94304,USA;~7 Department of Gynecology and Obstetrics,Guangdong Provincial People’s Hospital,Guangzhou510080.China. Acta Pharmacologica Sinica. 2007(11)
[7]Obesity and colorectal cancer risk: A meta-analysis of cohort studies[J]. Zhe Dai, Yan-Cheng Xu, Li Niu, Department of Endocrinology, Zhongnan Hospital, Wuhan University, Wuhan 430071, Hubei Province, China.. World Journal of Gastroenterology. 2007(31)
[8]细胞凋亡及其信号转导、凋亡抵抗与恶性肿瘤[J]. 夏金荣,赵东强. 临床荟萃. 2005(24)
[9]细胞凋亡过程中的信号转导[J]. 毛伟平,许锋. 环境与职业医学. 2004(01)
[10]细胞凋亡与大肠癌[J]. 洪道显. 现代诊断与治疗. 2002(01)
本文编号：2983438
【文章来源】：复旦大学上海市 211工程院校 985工程院校 教育部直属院校
【文章页数】：52 页
【学位级别】：硕士
【部分图文】：
凋亡调节通路
选用卜aetion，引物序列为(「):5一e丁ee奸eeAee仃eAee6仃ee一3，引物扩增长度为26sbp。扩增以94oC305，50℃305，72oCZmin为一循环，10min。图1为PcR产物的凝胶电泳扫描图像，第道是内参p一action，其位置在250bp偏上;第三oobp之间有一明显条带，与GHs一Rla大小相符有GHS一Rla的表达。
图2ghrelin对H下29细胞生长的影响注:与对照组比较，*p<0.05;与s一「u组比较，△P<0，05。OAPI染色典型的凋亡细胞核呈现染色质的浓缩、边集、核固缩、核碎裂，染色3中A、B、e依次是对照组、5一「uso协M处理组，s一「u+Gio一6M处理组，时间为24h。A图中细胞核基本都是正常形态;B图中可见核固缩、
【参考文献】：
期刊论文
[1]Different effects of ghrelin, des-acyl ghrelin and obestatin on gastroduodenal motility in conscious rats[J]. Mineko Fujimiya,Akihiro Asakawa,Koji Ataka,Ikuo Kato,Akio Inui. World Journal of Gastroenterology. 2008(41)
[2]Ghrelin and gastric acid secretion[J]. Koji Yakabi,Junichi Kawashima,Shingo Kato. World Journal of Gastroenterology. 2008(41)
[3]T-2 toxin-induced apoptosis involving Fas,p53,Bcl-xL,Bcl-2,Bax and caspase-3 signaling pathways in human chondrocytes[J]. Jing-hong CHEN Jun-ling CAO,Yong-lie CHU,Zhi-lun WANG,Zhan-tian YANG,Hong-lin WANG (Ministry of Education Key Laboratory of Environment and Genes related to Diseases,Institute of Endemic Diseases,School of Medicine,Xi’an Jiaotong University,Xi’an 710061,China). Journal of Zhejiang University（Science B:An International Biomedicine & Biotechnology Journal）. 2008(06)
[4]MAPKs在Bcl-2家族调控细胞凋亡中的作用[J]. 古晓娜,刘占旗,战景明,谢满廷. 国外医学(卫生学分册). 2008(02)
[5]PI3K-Akt信号通路与肿瘤细胞凋亡关系的研究进展[J]. 黄秀兰,崔国辉,周克元. 癌症. 2008(03)
[6]Oridonin induces apoptosis via PI3K/Akt pathway in cervical carcinoma HeLa cell line[J]. Hong-zhen HU~2,Yue-bo YANG~3,Xiang-dong XU~4,Hong-wei SHEN~5,Yi-min SHU~6,Zi REN~7,Xiao-mao LI~3,Hui-ming SHEN~3,Hai-tao ZENG~(3,8)~2 Department of Gynecology and Obstetrics,Yijishan Hospital,Wannan Medical College,Wuhu 241001,China;~3 Department of Gynecologyand Obstetrics,the Third Affiliated Hospital,Sun Yat-sen University,Guangzhou 510080,China;~4 Department of Surgery,the First AffiliatedHospital,Sun Yat-sen University,Guangzhou 510080,China;~2 Department of Gynecology and Obstetrics,the First Affiliated Hospital,SunYat-sen University,Guangzhou 510080.China;~6 Department of Obstetrics and Gynecology,IVF Program,Stanford University MedicalCenter,Palo Alto,CA 94304,USA;~7 Department of Gynecology and Obstetrics,Guangdong Provincial People’s Hospital,Guangzhou510080.China. Acta Pharmacologica Sinica. 2007(11)
[7]Obesity and colorectal cancer risk: A meta-analysis of cohort studies[J]. Zhe Dai, Yan-Cheng Xu, Li Niu, Department of Endocrinology, Zhongnan Hospital, Wuhan University, Wuhan 430071, Hubei Province, China.. World Journal of Gastroenterology. 2007(31)
[8]细胞凋亡及其信号转导、凋亡抵抗与恶性肿瘤[J]. 夏金荣,赵东强. 临床荟萃. 2005(24)
[9]细胞凋亡过程中的信号转导[J]. 毛伟平,许锋. 环境与职业医学. 2004(01)
[10]细胞凋亡与大肠癌[J]. 洪道显. 现代诊断与治疗. 2002(01)
本文编号：2983438
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