PTEN/NF-κB/Caspase信号通路对K562/ADM细胞阿霉素耐药逆转机制的研究

中国生物工程杂志

2013, Vol. 33

Issue (3): 54-60

研究报告

PTEN/NF-κB/Caspase信号通路对K562/ADM细胞阿霉素耐药逆转机制的研究

成志勇¹, 梁文同¹, 王素云², 颜晓燕¹, 李华¹, 王宝艳¹, 田赫¹, 魏玉涛¹, 芦希¹

1. 保定市第一医院保定 071000;
2. 河北省人民医院石家庄 050051

Mechanism of PTEN/NF-κB/Caspase Pathway on Adriamycin Resistance Reversal in K562/ADM Cells

CHENG Zhi-yong¹, LIANG Wen-tong¹, WANG Su-yun², YAN Xiao-yan¹, LI Hua¹, WANG Bao-yan¹, TIAN He¹, WEI Yu-tao¹, LU Xi ¹

1. The No1 Hospital of Baoding, Baoding 071000, China;
2. Hebei General Hospital, Shijiazhuang 050051, China

全文: PDF(903 KB) HTML

摘要： 目的:探讨PTEN/Akt/NF-κB信号通路对阿霉素耐药慢性粒细胞白血病细胞系K562/ADM细胞耐药逆转的分子作用机制。方法:将携带有野生型PTEN基因及绿色荧光蛋白的腺病毒(Ad-PTEN-GFP)或空载体(Ad-GFP)腺病毒转染K562/ADM细胞,在转染3d内联合应用不同浓度阿霉素(ADM),观察PTEN基因对阿霉素的协同抑制作用,根据IC₅₀计算药物逆转倍数。通过MTT检测细胞增殖,流式细胞术检测转染效率、细胞凋亡率,荧光定量PCR(FQ-PCR)检测 PTEN、NF-κB、I-κB、P53 基因水平变化,Western blot检测PTEN、Akt、p-Akt、P65蛋白质水平变化。结果:Ad-PTEN-GFP转染与阿霉素联合作用后,K562/ADM细胞对阿霉素的敏感性增加,细胞增殖明显受抑,凋亡率明显高于Ad-GFP与阿霉素联合作用组,耐药逆转倍数为3.8倍。与Ad-GFP组相比,Ad-PTEN-GFP 转染K562/ADM细胞3d后PTEN蛋白表达明显增加,p-Akt与P65蛋白表达明显下调,NF-κB mRNA表达水平下调,I-κB mRNA无明显改变,P53 mRNA表达轻度升高、Caspase-3/7活性增强。PTEN mRNA表达水平与NF-κB mRNA表达水平负相关(r=－0.968,P<0.05),NF-κB 蛋白活性与Caspase-3/7活性负相关(r=－0.986,P<0.05)。结论:野生型PTEN可能通过抑制PI3K/NF-κB/Caspase信号传导通路,逆转K562/ADM细胞的多药耐药。

关键词： PTEN; Akt; 耐药; 转录因子; NF-κB

Abstract: Objective:To investigate the mechanism of PTEN/Akt/NF-κB signal pathway in drug resistance reversal on K562/ADM cells in vitro. Methods: The recombinated adenovirus containing green fluorescent protein and PTEN(Ad-PTEN-GFP)or empty vector (Ad-GFP) was transfected into K562/ADM cells together with different concentration of adriamycin(ADM). We detected the different drug IC₅₀ with or without PTEN gene transfection to calculate the durg resistant factor (RF) by MTT; the proliferation of K562/ADM cells was observed by MTT assay; the apoptosis rate was assessed by flow cytometry (FCM). PTEN, NF-κB, I-κB, P53 mRNA were detected by real-time fluorescent relative-quantification reverse transcriptional PCR (FQ-PCR). PTEN, Akt, p-Akt and NF-κB protein levels were detected by western blot. Results: PTEN gene can enhance adriamycin sensitive in K562/ADM cells. The RF of ADM was 3.8 fold after transfection with PTEN gene. p-Akt and NF-Κb (P65) protein levels were down-regulated and NF-κB mRNA down-regulated too, but P53 up-regulated after transfaction with Ad-PTEN-GFP in K562/ADM cells, Caspase-3/7 activities up-regulated. PTEN mRNA expression levels were negative correlation with NF-κB mRNA(r=－0.968,P<0.05), NF-κB activities were negative correlation with Caspase-3/7 activities(r=－0.986,P<0.05). Conclusion: Wild PTEN gene might reverse adriamycin resistance via inhibiting PTEN/NF-κB/Caspase pathway.

Key words: PTEN PI3K/Akt Drug resistance Transcription factor NF-κB

收稿日期: 2012-10-08 出版日期: 2013-03-25

ZTFLH:

R733

通讯作者: 成志勇 E-mail: dzczy@sohu.com

	服务
	把本文推荐给朋友
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章
	王素云
	颜晓燕
	李华
	王宝艳
	田赫
	魏玉涛
	芦希
	成志勇
	梁文同

引用本文:

成志勇, 梁文同, 王素云, 颜晓燕, 李华, 王宝艳, 田赫, 魏玉涛, 芦希. PTEN/NF-κB/Caspase信号通路对K562/ADM细胞阿霉素耐药逆转机制的研究[J]. 中国生物工程杂志, 2013, 33(3): 54-60.

CHENG Zhi-yong, LIANG Wen-tong, WANG Su-yun, YAN Xiao-yan, LI Hua, WANG Bao-yan, TIAN He, WEI Yu-tao, LU Xi. Mechanism of PTEN/NF-κB/Caspase Pathway on Adriamycin Resistance Reversal in K562/ADM Cells. China Biotechnology, 2013, 33(3): 54-60.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2013/V33/I3/54

[1] Lin X, Zhang X, Wang Q,et al.Perifosine downregulates MDR1 gene expression and reverses multidrug-resistant phenotype by inhibiting PI3K/Akt/NF-κB signaling pathway in a human breast cancer cell line. Neoplasma. 2012;59(3):248-256.
[2] Han Z, Hong L, Han Y,et al. Phospho Akt mediates multidrug resistance of gastric cancer cells through regulation of P-gp, Bcl-2 and Bax. J Exp Clin Cancer Res. 2007;26(2):261-268.
[3] Morishita N, Tsukahara H, Chayama K, et al.Activation of Akt is associated with poor prognosis and chemotherapeutic resistance in pediatric B-precursor acute lymphoblastic leukemia. Pediatr Blood Cancer. 2012;59(1):83-89.
[4] 成志勇,张昕,王哲,等. PTEN和COX-2在髓系白血病中的表达及作用机制探讨.中华检验医学杂志,2012,35(2):165-169. Cheng Z Y, Zhang X, Wang Z. et al. The expression and mechanism of PTEN and COX-2 in myeloid leukemia cells. Chinese Journal of Laboratory Medicine, 2012,35(2):165-169.
[5] Vasudevan K M, Gurvmunhy S, Rangnekar V M. Suppression of PTEN expression by NF-kappaB prevents apoptosis. Mol Cell Biol, 2004,24(3):1007-1021.
[6] Jotta P Y, Ganazza M A, Silva A, et al.Negative prognostic impact of PTEN mutation in pediatric T-cell acute lymphoblastic leukemia. Leukemia. 2010,24(1):239-242.
[7] 成志勇,万建设,王亚丽,等. PTEN基因对慢性粒细胞白血病Survivin、Xiap、Smac调控的研究.中华医学杂志,2011,91 (40): 2868-2872. Cheng Z Y, Wan J S, Wang Y L. et al. Regulation of wild type PTEN gene on Survivin, Xiap and Smac in chronic leukemia cells. National Medical Journal of China, 2011,91 (40): 2868-2872.
[8] Shen S H, Gu L J, Liu P Q, et al.Comparative proteomic analysis of differentially expressed proteins between K562 and K562/ADM cells. Chin Med J. 2008;121(5):463-468.
[9] Tabellini G, Cappellini A, Tazzari P L,et al. Phosphoinositide 3-kinase/Akt involvement in arsenic trioxide resistance of human leukemia cells. J Cell Physiol. 2005, 202(2):623-634.
[10] Tabellini G, Tazzari P L, Bortul R,et al. Novel 2’-substituted, 3’-deoxy -phosphatidyl-myo-inositol analogues reduce drug resistance in human leukaemia cell lines with an activated phosphoinositide 3-kinase/Akt pathway.Br J Haematol. 2004,126(4):574-582.
[11] Grandage V L, Gale R E, Linch D C,et al. PI3-kinase/Akt is constitutively active in primary acute myeloid leukaemia cells and regulates survival and chemoresistance via NF-kappaB, Mapkinase and p53 pathways. Leukemia. 2005,19(4):586-594.
[12] Zhou M, Gu L, Findley H W, et al. PTEN reverses MDM2-mediated chemotherapy resistance by interacting with p53 in acute lymphoblastic leukemia cells. Cancer Res. 2003, 63(19):6357-6362.
[13] Liu F, Bardhan K, Yang D, et al. NF-κB directly regulates Fas transcription to modulate Fas-mediated apoptosis and tumor suppression. J Biol Chem. 2012 Jul 20;287(30):25530-25540. Bull Cancer. 2012;99(4):E49-54.
[14] Liu S Y, Liang Q J, Lin T X,et al.Lipopolysaccharide-enhanced early proliferation of insulin secreting NIT-1 cell is associated with nuclear factor-kappaB- mediated inhibition of caspase 3 cleavage. Chin Med J (Engl). 2011;124(22):3652-3656.

[1]	欧阳琴,李艳萌,徐安健,周冬虎,李振坤,黄坚. GTF2H2通过介导AKT信号通路影响肝癌细胞Hep3B的增殖和迁移^*[J]. 中国生物工程杂志, 2021, 41(6): 4-12.
[2]	董雪迎,梁凯,叶克应,周策凡,唐景峰. 受体酪氨酸激酶对自噬的调控及其研究进展^*[J]. 中国生物工程杂志, 2021, 41(5): 72-78.
[3]	陈玉琼,谭文华,刘海峰,陈根. miR-29a通过调控PTEN表达对脂多糖诱导人肺微血管内皮细胞损伤的保护作用研究^*[J]. 中国生物工程杂志, 2021, 41(5): 8-16.
[4]	董曙馨,秦磊,李春,李珺. 利用转录因子工程重塑代谢网络实现细胞工厂高效生产[J]. 中国生物工程杂志, 2021, 41(4): 55-63.
[5]	赵久梅,王哲,李学英. 调控软骨形成的信号通路及相关因子在骨髓间充质干细胞骨向分化中的作用^*[J]. 中国生物工程杂志, 2021, 41(10): 62-72.
[6]	王天柱,吴庆,张宁,王冬杰,许洲,罗伟,杜宗君. 鱼类黑色素合成及信号通路的研究进展^*[J]. 中国生物工程杂志, 2020, 40(5): 84-93.
[7]	戴奇男,张景红. 肿瘤多药耐药与自噬、DNA修复和肿瘤干细胞相关的分子机制研究进展 ^*[J]. 中国生物工程杂志, 2020, 40(4): 69-77.
[8]	户丽君,段亮,黄逸云,林璐,黄茂,陈露,彭棋,胡琴,张彦,周兰. S100A9参与介导具核梭杆菌促结肠癌细胞的增殖与迁移的作用 ^*[J]. 中国生物工程杂志, 2020, 40(1-2): 84-91.
[9]	赵建民,张思源. 耐药菌感染的噬菌体治疗专利技术[J]. 中国生物工程杂志, 2020, 40(10): 104-111.
[10]	张潘红,李莲莲,张秀美,崔家骏,姜银杰. microRNA对肺癌化疗耐药性影响的研究进展 ^*[J]. 中国生物工程杂志, 2019, 39(7): 79-84.
[11]	徐燕,刘正芸,张琬棂,王盛羽,王欢. 靶向干扰TAGLN表达对HBV阳性肝癌细胞生物学行为的影响及机制初探 ^*[J]. 中国生物工程杂志, 2019, 39(11): 13-21.
[12]	李文,陈洁,胡伟男,漆亚云,付毅红,刘佳敏,王贞超,欧阳贵平. EGFR耐药突变及其小分子抑制剂研究进展 ^*[J]. 中国生物工程杂志, 2019, 39(10): 97-104.
[13]	张晓勇,罗前程. 应用LNA-PCR法检测乙型肝炎病毒阿德福韦酯耐药位点基因突变 ^*[J]. 中国生物工程杂志, 2018, 38(9): 48-54.
[14]	赵悦,吴昊,乔建军. 细菌细胞壁生长调控机制研究进展 ^*[J]. 中国生物工程杂志, 2018, 38(8): 92-99.
[15]	叶中杨,邱怀雨,祝丙华,李泽,祝业,王立贵. sRNA调控细菌耐药相关基因表达研究进展 ^*[J]. 中国生物工程杂志, 2018, 38(7): 89-93.

Viewed

Full text

Abstract

Cited

Shared

Discussed