低氧诱导因子1α对子宫颈癌细胞生物学行为的影响  

Influence of hypoxia inducible factor-1α on cervical cancer cell line HeLa in vitro

作　　者：程艳香[1] 濮德敏[1] 刘嵘[1] 李天[1] 尹伶[1] 马丁[1] 

机构地区：[1]武汉华中科技大学同济医学院附属同济医院妇产科,430032

出　　处：《中华妇产科杂志》2007年第8期551-554,共4页Chinese Journal of Obstetrics and Gynecology

摘　　要：目的探讨低氧诱导因子1α(HIF-1α)对宫颈癌细胞的生物学行为的影响以及其中可能存在的分子机制。方法通过 CoCl_2化学诱导宫颈癌 HeLa 细胞缺氧;构建靶向 HIF-1α的反义真核表达载体、经脂质体介导转染 HeLa 细胞的方法沉默 HIF-1α的表达。将实验细胞分为常氧未转染对照(NN)组、常氧空质粒转染对照(NI)组、常氧转染 pcDNA3.0/HIF-1α质粒(NT)组、缺氧未转染对照(HN)组、缺氧空质粒转染对照(HT)组、缺氧转染 pcDNA3.0/HIF-1α质粒(HT)组。用四甲基偶氮唑蓝法、Transwell 侵袭小室方法观察各组细胞的增殖、侵袭能力的改变,用流式细胞仪检测各组细胞的凋亡率,用 RT-PCR 技术检测各组细胞目的基因 HIF-1α及其靶基因血管内皮生长因子(VEGF)、葡萄糖转运体1(GLUT1)、多药耐药基因1(MDR1)的表达变化。结果 NT 组细胞在培养12、24、48、72 h 时的活细胞数分别为0.053±0.003、0.074±0.004、0.148±0.015、0.192±0.038,而 HT 组分别为0.069±0.003、0.155±0.022、0.224±0.022、0.308±0.069;NT 和 HT 组的细胞增殖受到抑制,NT组与 NN 及 NI 组、HT 组与 HN 及 HI 组间的活细胞数分别比较,差异均有统计学意义(P<0.01)。各组细胞的凋亡率分别是,NN 组(29.27±0.18)%、NI 组(31.13±0.08)%、NT 组(51.11±0.14)%、HN 组(11.46±0.28)%、HI 组(15.77±0.49)%、HT 组(40.05±0.97)%;HT 组与 HN 及 HI 组、NT组与 NN 及 NI 组间分别比较,差异均有统计学意义(P<0.01)。各组细胞的侵袭能力由高到低依次为 HI、HN、NI、NN、HT、NT 组,分别为(40±9)%、(37±12)%、(28±5)%、(26±7)%、(19±7)%、(10±5)%;NT 组与 NN 及 NI 组、HT 组与 HN 及 HI 组间分别比较,差异均有统计学意义(P<0.05)。NT 组细胞 HIF-1α、VEGF、GLUT1、MDR1 mRNA 的相对表达量分别为0.05±0.12、0.09±0.11、0.08±0.15、0.05±0.15,而 HT 组分别为0.04±0.16、0.16±0.16、0.12±0.20、0.20±0.21;NT 组与 NN及 NI 组、HT 组与 HN 及 HI 组间分别比较,HIObjective To explore the direct influence of hypoxia inducible factor-1α （HIF-1α） on the development of invasive cervical cancer and the possible molecular mechanism. Methods Recombinant antisense targeting HIF-1α eukaryotic expression vector was constructed and transfected into cultured human cervical cancer cell line HeLa to reduce the expression of HIF-1α and its effect on cell proliferation, apoptosis, invasion and the cascade downstream gene expression of HIF-1α, including vascular endothelial growth factor （VEGF）, glucose transport 1 （GLUT1） and multidrug resistance 1 （MDR1） genes was observed. The chemical method using CoCl2 to induce hypoxia environment of growing ceil was performed. Ceils were divided into six groups, NN （normal non-transfected）, NI （normal invalid transfected）, NT （ normal transfected）, HN （ hypoxia non-transfected）, HI （ hypoxia invalid transfected）, and HT （ hypoxia transfected）. Methyl thiazolyl tetrazolium （MTT）, flow cytometry, and Transwell methods were performed to evaluate the proliferation, invasion and apoptosis, and RT-PCR method was used to detect the gene expression of HIF-1α, VEGF, GLUT1 and MDR1. Results After induction of hypoxia by COCl2, the change of gene expression of HIF-1α in HN （or HI） group compared to that in NN （ or NI） group was not obvious （P 〉 0. 05 ）, but expression of VEGF, GLUT1 and MDR1 were all enhanced and overall proliferation was promoted, apoptosis inhibited [ （ 11.46 ± 0. 28）% vs （29. 27 ± 0. 18）% , （ 15.77 ± 0.49）% vs （31.13±0.08）%], and transmembrane behavior enhanced [（37 ±12）% vs （26±7）%, （40 ±9）% vs （28 ±5）% ], and the variations were significant （P 〈0. 05）. On the contrary, transfection with pcDNA3.0/HIF-1α was companied by declined gene expression of HIF-1α （ NT： 0. 05 ± 0. 12, HT： 0.04 ± 0. 16）, and all the variations were significant （ P 〈 0. 05）. Conclusions HIF-1α may participate in malignant biological behavi

关 键 词：宫颈肿瘤 DNA结合蛋白质类 核蛋白质类 血管内皮生长因子A 单糖转运蛋白质类 基因 MDR 

分 类 号：R737.33[医药卫生—肿瘤]