缺氧及HIF-1a对结直肠癌上皮-间质转化及侵袭增殖的影响

目的研究不同缺氧条件下缺氧诱导因子-1a（HIF-1a）、锌指转录因子Snail及上皮钙黏蛋白（E-cadherin）在结直肠癌SW480细胞中的表达水平，以及它们对细胞的增殖凋亡和侵袭迁移力的影响，探讨三者在结直肠癌的缺氧及上皮间质转化中的作用及可能机制。方法利用氯化钴（CoCl2）化学诱导细胞缺氧模型，实验分组：常氧组、低氧组和无氧组。采用MTT法检测结直肠癌SW480细胞的生长增殖活性，体外侵袭实验（Transwell小室法）检测细胞的侵袭迁移力，流式细胞术（FCM）检测细胞的凋亡及周期变化，RT—PCR及Westernblot检测细胞中HIF-1a、Snail及E-cadherinmR...

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Bibliographic Details
Published in肿瘤防治研究 Vol. 39; no. 10; pp. 1170 - 1175
Main Author 周明利 卢先州 张留柱 向晶 谢荣俊 肖帅 张树友
Format Journal Article
LanguageChinese
Published 2012
Subjects
Snail
上皮-间质转化
上皮钙黏蛋白
缺氧
缺氧诱导因子-1工
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ISSN1000-8578

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Summary:目的研究不同缺氧条件下缺氧诱导因子-1a（HIF-1a）、锌指转录因子Snail及上皮钙黏蛋白（E-cadherin）在结直肠癌SW480细胞中的表达水平，以及它们对细胞的增殖凋亡和侵袭迁移力的影响，探讨三者在结直肠癌的缺氧及上皮间质转化中的作用及可能机制。方法利用氯化钴（CoCl2）化学诱导细胞缺氧模型，实验分组：常氧组、低氧组和无氧组。采用MTT法检测结直肠癌SW480细胞的生长增殖活性，体外侵袭实验（Transwell小室法）检测细胞的侵袭迁移力，流式细胞术（FCM）检测细胞的凋亡及周期变化，RT—PCR及Westernblot检测细胞中HIF-1a、Snail及E-cadherinmRNA与蛋白的表达。结果MTT结果显示随CoCl2浓度的增加结直肠癌SW480细胞的增殖活性逐渐增加（P〈0．05）；Transwell结果显示随缺氧程度的增加穿过基质膜和聚碳酸酯膜的细胞数逐渐增多，侵袭迁移率增加（P〈0．05）；FCM检测结果显示与常氧组比较，无氧组和低氧组G0／G1期SW480细胞明显减少，G2／S期细胞明显增多，细胞凋亡率降低（P〈0．05）；RT—PCR与Westernblot检测结果显示低氧组、无氧组与常氧组比较，SW480细胞内HIF-1a和SnailmRNA与蛋白的表达增加，E—cadherinmR—NA和蛋白的表达降低（P〈0．05），且HIF-1a和Snail的表达呈显著正相关，HIF-1a、Snail与E—cadherin的表达均呈显著负相关（P〈0．05）。结论缺氧及HIF—1a 可促进结直肠癌SW480细胞株的体外生长增殖活性，抑制其凋亡，增强其体外侵袭迁移能力，其机制可能是缺氧诱导HIF—1a表达上调直接作用于SW480细胞产生，亦可能是缺氧或HIF-1a表达上调通过促进Snail表达，抑制E—cadherin表达，间接导致EMT的发生发展而产生。
Bibliography:42-1241/R
Objective To study the expression level of the hypoxia inducible factor-1 a （HIF1a） ,zinc finger transcription factor Snail and E-cadherin in different hypoxia condition in the colorectal cancer SW480 cells,and to explore their effect on the proliferation, apoptosis, invasion and transference, approach the roles and mechanisms of them three in the hypoxia and epithelial-mesenehymal transition of colorectal cancer. Methods Chemical was used to induce the cell hypoxia model by cobalt chloride（CoCl2 ）, and three groups：normoxia group, hypoxia group and non-oxygen group were involved in this study. Methyl thiazolyl tetrazolium（MTT） method was used to detect the proliferative of colorectal cancer SW480 cells, extraorgan invasion experiment（Tran,;well method） was applied to evaluate the invasion and transference capability of cells,the cell cycle and apoptosis was examined by flow eytometry（FCM）,the mRNA and protein expression levels of HIF-1a,Snail and E-cadherin were detected by real time PCR（RT-PCR） and
ISSN:1000-8578