常氧条件下调控表达缺氧诱导因子1α对肝癌细胞增殖和侵袭能力的影响  

Inducible expression of hypoxia-inducible factor 1α on the proliferation and invasion property of HepG2 cells under normoxia in vitro

作　　者：金炜东[1] 陈孝平[1] 杨盛利[1] 徐宗全[1] 张万广[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院肝脏外科

出　　处：《中华外科杂志》2007年第23期1634-1636,共3页Chinese Journal of Surgery

基　　金：国家自然科学基金(30371395)

摘　　要：目的探讨在体外常氧条件下,诱导表达缺氧诱导因子1α(HIF-1α)对肝癌细胞(HepG2)增殖及侵袭能力的影响。方法利用 Tet-on 基因调控系统构建能诱导表达 HIF-1α的HepG2^(Tet-on-HIF-1α)细胞系;常氧条件下,噻唑兰法检测 HIF-1α对细胞增殖、黏附能力的影响,Transwell 法检测其对 HepG2细胞侵袭能力的影响。结果常氧条件下,强力霉素(1μg/ml)可诱导HepG2^(Tet-on-Hlf-1α)细胞 HIF-1αmRNA 和蛋白表达增加;增殖实验中,Dox(+)组与 Dox(-)组各时段吸光度 A_(490nm)值无差异(P>0.05);黏附实验中,Dox(+)组的 A_(490nm)值明显高于 Dox(-)组(P=0.008);Dox(+)组侵袭细胞数[(37.611±8.424)个]明显高于 Dox(-)组[(25.333±8.117)个](P<0.01)。结论 Tet-on 基因调控系统可上调 HIF-1α mRNA 的转录并增加其蛋白的表达;常氧条件下,HIF-1α不影响 HepG2细胞的增殖,但明显增加其黏附和侵袭能力。Objective To study the inducible expression of hypoxia-inducible factor -1α （HIF-1α） on the proliferation and invasion property of HepG2 cells under normoxia in vitro. Methods Constructed the 2HepG2^Tet-on-HIF-1α cell line which could induce the expression of HIF--1α by doxycycline; Under normoxia in vitro, MTT assay was used to observe the proliferative and adhesive activity of cells, and the invasive activity was determined by transwell cell culture chamber method. Results Under normoxia, the HIF--1α mRNA and protein of HepG2^Tet-on-HIF-1α cells could be induced up to （ 5. 899 ± 2. 176 ） and （ 2. 179 ± 0. 742 ） folds by doxycycline（ 1 μg/ml） ; There were no difference of A490nm between the Dox （ ＋ ） and Dox （ - ） group in experiment detecting the proliferation activity （ P 〉0. 05 ） ; But in adhesive experiment, the A490nm of Dox （ ＋ ）group was 0. 662 ±0. 058, higher than the Dox（ - ） group O. 526 ±0. 808 （P=0. 008） ; The invasived cell number of Dox（ ＋ ）group was 37. 611 ± 8.424,but in the Dox（ - ）group,the number was 25. 333 ± 8. 117 （P 〈0. 01 ）. Conclusions Under normoxia in vitro, the Tet-on gene regulate system could increase the HIF--1α protein by inducing the HIF--1α mRNA; HIF--1α has no influence with the proliferation activity, but it could enhance the adhesive and invasive properties of HepG2 cells.

关 键 词：肝肿瘤 实验性 基因表达调控 肿瘤 缺氧诱导因子1Α 增殖 侵袭 

分 类 号：R735.7[医药卫生—肿瘤]