| PTEN基因表达改变对人气道平滑肌迁移的影响 |
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| 引用本文： | 高杨,钟浩海,罗雅玲,赖文岩,徐健,张达成,蓝海兵.PTEN基因表达改变对人气道平滑肌迁移的影响[J].南方医科大学学报,2011,31(3):403-408. |
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| 作者姓名： | 高杨 钟浩海 罗雅玲 赖文岩 徐健 张达成 蓝海兵 |
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| 作者单位： | 高杨,罗雅玲,蓝海兵,GAO Yang,LUO Ya-ling,LAN Hai-bing(南方医科大学南方医院呼吸内科,广东,广州,510515);钟浩海,ZHONG Hao-hai(惠州市中心医院呼吸科,广东,惠州516000);赖文岩,LAI Wen-yan(南方医科大学南方医院心内科,广东,广州,510515);徐健,XU Jian(南方医科大学南方医院肾移植科,广东,广州,510515);张达成,ZHANG Da-cheng(南方医科大学华瑞医院呼吸内科,广东,广州,510630) |
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| 基金项目： | 国家自然科学基金，广东省自然科学基金 |
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| 摘 要： | 目的通过腺病毒载体体外转染人气道平滑肌细胞(HASMC),使肿瘤抑制基因PTEN过表达和RNA干扰表达,观察其对HASMC迁移的影响,并探讨其作用机制。方法利用携带野生型PTEN基因的重组腺病毒(Ad-GFP-PTEN)和针对PTEN基因的腺病毒短发夹状RNA(shRNA)载体(Ad-GFP-shRNA-PTEN)转染HASMC细胞,并以感染空载腺病毒Ad-GFP和空白组(DMEM)作为对照,用流式细胞计数确立最佳转染复数后,通过Transwell趋化小室和细胞划痕实验检测HASMC跨膜迁移能力和横向迁移能力的变化。Western blotting检测PTEN蛋白的表达和AKT、ERK1/2通路的活化情况,并加入两通路的抑制剂作为阳性对照。结果腺病毒搭载的PTEN基因过表达和干扰载体能成功改变PTEN基因的表达。上调PTEN表达能显著抑制HASMC迁移;蛋白水平上抑制PI3K/AKT通路,而MAPK/ERK通路未见变化。下调PTEN表达不能明显促进HASMC迁移;但在蛋白水平能使PI3K/AKT通路激活,而MAPK/ERK通路却受到抑制。结论上调PTEN表达能有效抑制HASMC的迁移,可能主要是通过抑制PI3K/AKT通路起作用。
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| 关 键 词： | PTEN 人气道平滑肌 迁移 PI3K/AKT 哮喘 |
Alteration of PTEN gene expression affects the migration of human airway smooth muscle cells in vitro |
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| GAO Yang,ZHONG Hao-hai,LUO Ya-ling,LAI Wen-yan,XU Jian,ZHANG Da-cheng,LAN Hai-bing.Alteration of PTEN gene expression affects the migration of human airway smooth muscle cells in vitro[J].Journal of Southern Medical University,2011,31(3):403-408. |
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| Authors: | GAO Yang ZHONG Hao-hai LUO Ya-ling LAI Wen-yan XU Jian ZHANG Da-cheng LAN Hai-bing |
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| Institution: | GAO Yang1,ZHONG Hao-hai2,LUO Ya-ling1,LAI Wen-yan3,XU Jian4,ZHANG Da-cheng5,LAN Hai-bing11Department of Respiratory Diseases,3Department of Cardiology,4Department of Kidney Transplantation,Nanfang Hospital,Southern Medical University,Guangzhou 510515,China,2Department of Respiratory Diseases,Huizhou Central Hospital,Huizhou 516000,5Department of Respiratory Diseases,Huarui Hospital,Guangzhou 510630,Chin |
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| Abstract: | Objective To investigate the changes in human airway smooth muscle cell(HASMC) migration and related signaling pathway after interference with PTEN gene expression.Method HASMCs were infected with an adenovirus vector and RNA interference vector of human PTEN gene to establish the cell model with PTEN gene over-expression(Ad-GFP-PTEN-HASMC) and one with PTEN gene silencing(Ad-shPTEN-HASMC),using Ad-GFP-infected and a blank cells as the negative controls and LY294002 as the positive control.Fluorescence micr... |
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| Keywords: | PTEN P13K/AKT |
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