[1]杨庆强,张才全,唐华.低氧对人结肠癌细胞HT-29体外侵袭转移潜能的影响[J].陆军军医大学学报(原第三军医大学学报),2007,29(23):2257-2260.
 YANG Qing-qiang,ZHANG Cai-quan,TANG Hua.Effects of moderate hypoxia on invasion and metastasis of colon carcinoma cell line HT-29 in vitro[J].J Amry Med Univ (J Third Mil Med Univ),2007,29(23):2257-2260.
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低氧对人结肠癌细胞HT-29体外侵袭转移潜能的影响(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
29卷
期数:
2007年第23期
页码:
2257-2260
栏目:
论著
出版日期:
2007-12-15

文章信息/Info

Title:
Effects of moderate hypoxia on invasion and metastasis of colon carcinoma cell line HT-29 in vitro
作者:
杨庆强张才全唐华
重庆医科大学附属第一医院普通外科
Author(s):
YANG Qing-qiang ZHANG Cai-quan TANG Hua
Department of General Surgery, The First Affiliated Hospital, Chongqing Medical University
关键词:
低氧骨桥蛋白核转录因子恶性转化
Keywords:
moderate hypoxia osteopontin nuclear factor kappa B malignant transformation
分类号:
R364.4; R73-37; R735.35
文献标志码:
A
摘要:
目的  探讨低氧与人结肠癌细胞HT-29恶性转化的关系,检测骨桥蛋白(osteopontin,OPN)、细胞核内核转录因子(NFκB/p65)表达和MMP-2、MMP-9活性变化,以探索其恶性转化的潜在机制。  方法  参照体内肿瘤的氧分压,建立低氧模型。MTT比色试验测定HT-29细胞的异质性黏附能力,Transwell侵袭试验判定其侵袭游走能力;将HT-29细胞低氧处理0、6、12、24、48 h,明胶酶谱分析检测分泌的MMP-2、MMP-9活性变化,RT-PCR和Western blot检测OPN mRNA和蛋白表达,提取细胞核蛋白,Western blot检测细胞核内NF-κB/p65蛋白表达。  结果  低氧诱导24 h后,HT-29细胞的异质性黏附能力、侵袭游走能力显著增加。低氧下6 h,MMP-2、MMP-9活性无明显变化,随着低氧时间的延长,其活性逐渐显著上调,24 h达顶峰,48 h与24 h无显著性差异。HT-29细胞在低氧的诱导下,其OPN mRNA和蛋白以及细胞核内NF-κB/p65蛋白的表达趋势与MMP-2、MMP-9活性变化趋势类同。  结论  低氧能诱导HT-29细胞异质性黏附能力、侵袭游走能力显著增加,MMP-2、MMP-9活性上调而促使其向恶性表型转化。OPN-NF-κB可能是低氧下继HIF-1之外的另一重要调节途径,该途径与低氧诱导的恶性表型有密切关系。
Abstract:
Objective To investigate the correlation between moderate hypoxia and maliglant transformation of colon carcinoma cell line HT-29.  Methods  Hypoxic conditions were produced according to p(O2) of tumor in vivo. The heterotypic adhesiveness of HT-29 cells was detected by MTT assay. Their reaction to the hypoxic environment was determined via invasion across a Matrigel-coated Transwell filter. At 0, 6, 12, 24 and 48 h after hypoxia, MMP-2/9 activities were assessed by gelatin zymography, and osteopontin mRNA and protein levels by RT-PCR and Western blot respectively. NF-κB/p65 levels in nucleus was measured by Western blot after extraction of nucleic protein. Results After 24-hour moderate hypoxia, the heterotypic adhesiveness and invasive capacity of HT-29 cells were significantly enchanced. MMP-2, MMP-9 activities under moderate hypoxia for 6 h showed no significant change as compared with that under normoxia, then gradually up-regulated and reached the peak at 24 h. They showed no significant difference between 24 h and 48 h after hypoxia. The expression trend of osteopontin mRNA, protein and NF-κB/p65 protein in nucleus was similar to that of MMP-2, MMP-9 under moderate hypoxia. Conclusion Moderate hypoxia could induce maliglant transformation in HT-29 cells through enhancing heterotypic adhesiveness and invasive capacity and upregulating MMP-2, MMP-9 activities. OPN-NF-κB may be an important regulatory pathway besides HIF-1 under hypoxic condition, which involves maliglant phenotype induced by moderate hypoxia.

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更新日期/Last Update: 2008-05-23