[1]石岩岩,丁士刚,鲁凤民,等.硫氧还蛋白1对胃癌细胞系BCG823生长的影响[J].中国微创外科杂志,2011,11(11):1030-1033.
　Shi Yanyan*,Ding Shigang*,Lu Fengmin,et al.Effects of Human Thioredoxin1 on the Growth of Gastric Cancer Cell BCG823[J].Chinese Journal of Minimally Invasive Surgery,2011,11(11):1030-1033.
点击复制

硫氧还蛋白1对胃癌细胞系BCG823生长的影响()
分享到：

《中国微创外科杂志》[ISSN:1009-6604/CN:11-4526/R]

卷:
11
期数:
2011年11期
页码:
1030-1033
栏目:
基础研究
出版日期:
2011-11-20

文章信息/Info

Title:
Effects of Human Thioredoxin1 on the Growth of Gastric Cancer Cell BCG823
作者:
石岩岩; 丁士刚; 鲁凤民; 张婷①; 张静.刘琳娜; 王晔
北京大学第三医院消化科，北京100191
Author(s):
Shi Yanyan*;  Ding Shigang*;  Lu Fengmin;  et al.
*Department of Gastroenterology, Peking University Third Hospital, Beijing 100191, China
关键词:
硫氧还蛋白1; 重组载体; 细胞周期; BCG823
Keywords:
Thioredoxin1; Recombinant vector; Cell cycle; BCG823
分类号:
R329.2+6
文献标志码:
A
摘要:
目的探讨硫氧还蛋白1（thioredoxin1, Trx1）对胃癌细胞系细胞周期及增殖速度的影响。方法对BCG823、AGS、MNK45、NUGC3、PHM82五种胃癌细胞系的质粒转染效率进行测定，选择出转染效率高的细胞系；并用实时PCR（RTPCR）测定胃癌细胞系中Trx1的表达水平，选择出表达较低的细胞系。将所构建的Trx1真核重组表达载体pcDNA3.1mycHisTrx1转染入上述实验筛选出的胃癌细胞系，使细胞中Trx1过表达，用流式细胞仪检测过表达Trx1的胃癌细胞进入S期的比例。结果BCG823、AGS两种细胞系的转染效率高，满足实验需要。BCG823的Trx1表达较AGS低 ［BCG823细胞系的ΔCT (TrxGAPDH)=1.43，AGS细胞系的ΔCT(TrxGAPDH)=1.10，ΔCT值与表达量成反比］，选择BCG823作为实验对象。转染pcDNA3.1mycHisTrx1的细胞Trx1表达升高（与内参蛋白的比值为0.45±0.02和0.26±003，n=3，t=9.127，P=0.000），进入S期的细胞比例升高（转染带有Trx1质粒的细胞进入S期的比例为28.54%，而转染了空载体者进入S期的比例为22.24%， χ2=104.759,P=0.000）。结论Trx1表达水平上调可以促进胃癌细胞进入S期。
Abstract:
ObjectiveTo study the effects of human thioredoxin1 (Trx1) on the cell cycle and proliferation of gastric cancer cells.MethodsTransfection efficiency of five gastric cancer cell line, BCG823, AGS, MNK45, NUGC3, and PHM82 was analyzed to choose the one with highest efficiency for our experiment. At the same time, the expression of Trx1 in the gastric cancer cell lines was measured by realtime PCR, so that to select the cell lines with lowest expression rate, which could avoid the interference from the endogenous Trx1 to the transfected Trx1, and to ensure the obvious effects of transfection. Then, the pcDNA3.1mycHisTrx1 recombinant vector was transfected into the cell line selected to make the Trx1 enhanced. The percentage of the cells in S phase was detected by flow cytometry. ResultsThe transfection efficiency of BCG823 and AGS was high enough. The Trx1 expression in BCG823 was lower than AGS ［ΔCT (TrxGAPDH) of BCG823 =1.43, ΔCT (TrxGAPDH) of AGS= 1.10, and ΔCT is reverse to the level of Trx expression］, so BCG823 was selected. The expression of Trx1 was upregulated by transfecting with pcDNA3.1mycHisTrx1 (the ratios to the referent protein are relatively 0.45±0.02 and 0.26±0.03, n=3, t=9.127, P=0.000). The percentage of the cells in S phase was significantly increased (after interfecting by the plasmid with Trx1, the rate of cells coming into S phase was 28.54%, compared with the rate of the cells interfected by the plasmid without Trx1, which was 22.24%, χ2=104.759，P=0.000). ConclusionsThe upregulation of Trx1 can promote the entry of BCG823 cells into S phase.

参考文献/References:

[1]Kaimul AM,Nakamura H,Masutani H,et al.Thioredoxin and thioredoxinbinding protein2 in cancer and metabolic syndrome.Free Radic Biol Med,2007,43:861-868.
[2]Becana M,Matamoros MA,Udvardi M,et al.Recent insights into antioxidant defenses of legume root nodules.New Phytol,2010,188: 960-976.
[3]Lillig CH,Holmgren A.Thioredoxin and related moleculesfrom biology to health and disease.Antioxid Redox Signal,2007,9:25-47.
[4]Koc A,Mathews CK,Wheeler LJ,et al.Thioredoxin is required for deoxyribonucleotide pool maintenance during S phase.J Biol Chem,2006,281:15058-15063.
[5]石岩岩，丁士刚，蒋素贞，等.人硫氧还蛋白1基因克隆及其重组载体的构建与鉴定.山东大学学报（医学版）,2010,48:41-45.
[6]Ceccarelli J,Delfino L,Zappia E,et al.The redox state of the lung cancer microenvironment depends on the levels of thioredoxin expressed by tumor cells and affects tumor progression and response to prooxidants.Int J Cancer,2008,123:1770-1778.
[7]Shan W,Zhong W,Zhao R,et al.Thioredoxin 1 as a subcellular biomarker of redox imbalance in human prostate cancer progression.Free Radic Biol Med,2010,49:2078-2087.
[8]Noda N,Ochiai A,Miyazaki K,et al.Detection of thioredoxin in gastric cancer: association with histological type.Antioxid Redox Signal,2000,2:519-528.
[9] Lincoln DT,AlYatama F,Mohammed FM,et al.Thioredoxin and thioredoxin reductase expression in thyroid cancer depends on tumor aggressiveness.Anticancer Res,2010,30:767-775.
[10]Jones DP.Redox sensing:orthogonal control in cell cycle and apoptosis signalling.J Intern Med,2010,268:432-448.
[11]Young MG,Thoma RZ,Jennifer MJ,et al.Selective protection of nuclear thioredoxin1 and glutathione redox systems against oxidation during glucose and glutamine deficiency in human colonic epithelial cells.Free Radic Biol Med,2007,42:367-370.
[12]Baker LM,Raudonikiene A,Hoffman PS,et al.Essential thioredoxindependent peroxiredoxin system from Helicobacter pylori: genetic and kinetic characterization.J Bacteriol,2001,183:1961-1973.
[13]Yoshioka J,Schreiter ER,Lee RT.Role of thioredoxin in cell growth through interactions with signaling molecules.Antioxid Redox Signal,2006,8:2143-2151.
[14]Mochizuki M,Kwon YW,Yodoi J,et al.Thioredoxin regulates cell cycle via the ERK1/2cyclin D1 pathway.Antioxid Redox Signal,2009,11:2957-2971.

备注/Memo

备注/Memo:
基金项目：国家自然科学基金资助项目（30770980）丁士刚通讯作者，Email：dingshigang222@163.com鲁凤民（北京大学医学部病原生物学系，北京100191)
更新日期/Last Update: 2013-05-06