[1]罗勤利,邓小垭,董飞,等.RNA结合蛋白TTP对肺腺癌细胞增殖影响的实验研究[J].第三军医大学学报,2019,41(09):851-858.
 LUO Qinli,DENG Xiaoya,DONG Fei,et al.RNA binding protein tristetraprolin inhibits proliferation of lung adenocarcinoma cells in vitro[J].J Third Mil Med Univ,2019,41(09):851-858.
点击复制

RNA结合蛋白TTP对肺腺癌细胞增殖影响的实验研究(/HTML )
分享到:

《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
41卷
期数:
2019年第09期
页码:
851-858
栏目:
基础医学
出版日期:
2019-05-15

文章信息/Info

Title:
RNA binding protein tristetraprolin inhibits proliferation of lung adenocarcinoma cells in vitro
作者:
罗勤利邓小垭董飞徐莉江涛
重庆医科大学附属第一医院呼吸与危重症医学科
Author(s):
LUO Qinli DENG Xiaoya DONG Fei XU Li JIANG Tao

Department of Respiratory and Critical Care Medicine, the First Affiliated Hospital of Chongqing Medical University, Chongqing, China

关键词:
RNA结合蛋白肺腺癌凋亡细胞增殖
Keywords:
RNA binding protein lung adenocarcinoma apoptosis cell proliferation
分类号:
R341; R730.23; R734.2
文献标志码:
A
摘要:

目的探讨TTP(tritetraprolin)过表达对肺腺癌细胞增殖影响及其可能的作用机制。方法构建带有四环素可调控(tetracyclineon,Teton)的TTP基因表达系统,利用慢病毒感染的方法转染肺腺癌H1975及1299细胞,并在强力霉素(doxycycline,Dox)的诱导下稳定表达TTP。用实时荧光定量PCR和Western blot鉴定TTP受Dox调控表达的效果。分别采用CCK8法、Transwell、平板克隆形成实验、流式细胞术检测TTP过表达后对细胞增殖、迁移、集落形成能力、凋亡和细胞周期的影响。Western blot法检测TTP过表达对凋亡基因在蛋白水平表达的影响。结果成功构建受Dox诱导调控的TTP过表达肺腺癌细胞模型。与TTP低表达组比较,Dox诱导TTP过表达组明显抑制H1975、H1299细胞增殖、迁移能力(H1975 :P<0.01,H1299: P<0.001)及集落形成能力(P值均<0.0001),且使H1975细胞周期被阻滞在S期(P<0.01),但TTP过表达不影响肺腺癌细胞凋亡。结论TTP过表达显著抑制肺腺癌细胞增殖、迁移、集落形成能力,使细胞周期被阻滞在S期,但TTP过表达不通过细胞凋亡途径影响肺腺癌细胞增殖和生长。

Abstract:

ObjectiveTo investigate the effect of tristetraprolin (TTP) on the proliferation of lung cancer cells and to explore the possible mechanism. MethodsThe human lung adenocarcinoma cells with tetracyclineinducible (Teton) expression regulatory system were established in H1975 and 1299 cells with the aid of lentivirus infection. Western blotting and qPCR were used to confirm the overexpression of TTP in the cells after the doxycycline (Dox) treatment. Effect of TTP on the proliferation, migration and colonyforming capacity were detected by CCK-8 assay, Transwell test and colony forming unit test respectively. The cell apoptosis and cell cycle were measured by flow cytometry. The expression of cellular apoptosis-related proteins were determined by Western blotting. ResultsLung adenocarcinoma cells with Doxinduced TTP overexpression were successfully established. Compared with the cells with low TTP expression, those with Dox-induced TTP overexpression siginicantly inhibited proliferation, migration(H1975: P<0.01, H1299: P<0.001)and colony-formation (all P<0.0001) in lung adenocarcinoma cells H1975 and H1299. What’s more, H1975 cells were arrested at S stage (P<0.01). However, TTP overexpression exerted no effect on cell apoptosis. ConclusionTTP overexpression obviously inhibits proliferation, migration and colonyforming capacity, and arrests cell cycle at S stage in lung adenocarcinoma cells. But the mechanism is not due to the regulation of cell apoptosis.

参考文献/References:

[1]OGILVIE R L, ABELSON M, HAU H H, et al. Tristetraprolin downregulates IL2 gene expression through AUrich elementmediated mRNA decay[J]. J Immunol, 2005, 174(2): 953-961. DOI:10.4049/jimmunol.174.2.953.
[2]WEI Z R, LIANG C, FENG D, et al. Low tristetraprolin expression promotes cell proliferation and predicts poor patients outcome in pancreatic cancer[J]. Oncotarget, 2016, 7(14): 17737-17750. DOI:10.18632/oncotarget.7397.
[3]GODDIO M V, GATTELLI A, SLOMIANSKY V, et al. Mammary differentiation induces expression of tristetraprolin, a tumor suppressor AUrich mRNAbinding protein[J]. Breast Cancer Res Treat, 2012, 135(3): 749-758. DOI:10.1007/s1054901222160.
[4]HALEES A S, ELBADRAWI R, KHABAR K S. ARED Organism: expansion of ARED reveals AUrich element cluster variations between human and mouse[J]. Nucleic Acids Res, 2008, 36(Database issue): D137-D140. DOI:10.1093/nar/gkm959.
[5]BLACKSHEAR P J. Tristetraprolin and other CCCH tandem zincfinger proteins in the regulation of mRNA turnover[J]. Biochem Soc Trans, 2002, 30(Pt 6): 945-952. DOI:10.1042/.
[6]BAOU M, JEWELL A, MUTHURANIA A, et al. Involvement of Tis11b, an AUrich binding protein, in induction of apoptosis by rituximab in B cell chronic lymphocytic leukemia cells[J]. Leukemia, 2009, 23(5): 986-989. DOI:10.1038/leu.2008.340.
[7]GUO J, QU H, CHEN Y, et al. The role of RNAbinding protein tristetraprolin in cancer and immunity[J]. Med Oncol, 2017, 34(12): 196. DOI:10.1007/s1203201710556.
[8]JELTSCH K M, HEISSMEYER V. Regulation of T cell signaling and autoimmunity by RNAbinding proteins[J]. Curr Opin Immunol, 2016, 39: 127-135. DOI:10.1016/j.coi.2016.01.011.
[9]FALLAHI M, AMELIO A L, CLEVELAND J L, et al. CREB targets define the gene expression signature of malignancies having reduced levels of the tumor suppressor tristetraprolin[J]. PLoS ONE, 2014, 9(12): e115517. DOI:10.1371/journal.pone.0115517.
[10]FU M, BLACKSHEAR P J. RNAbinding proteins in immune regulation: a focus on CCCH zinc finger proteins[J]. Nat Rev Immunol, 2017, 17(2): 130-143. DOI:10.1038/nri.2016.129.
[11]BROOKS S A, BLACKSHEAR P J. Tristetraprolin (TTP): interactions with mRNA and proteins, and current thoughts on mechanisms of action[J]. Biochim Biophys Acta, 2013, 1829(6/7): 666-679. DOI:10.1016/j.bbagrm.2013.02.003.
[12] GRUBER A R, FALLMANN J, KRATOCHVILL F, et al. AREsite: a database for the comprehensive investigation of AUrich elements[J]. Nucleic Acids Res, 2011, 39(Database issue): D66-D69. DOI:10.1093/nar/gkq990.
[13]KHABAR K S A. Hallmarks of cancer and AUrich elements[J]. Wiley Interdiscip Rev: RNA, 2016, 8(1): e1368. DOI:10.1002/wrna.1368.
[14]BISOGNO L S, KEENE J D. RNA regulons in cancer and inflammation[J]. Curr Opin Genet Dev, 2018, 48: 97-103. DOI:10.1016/j.gde.2017.11.004.
[15]MUKHERJEE N, JACOBS N C, HAFNER M, et al. Global target mRNA specification and regulation by the RNAbinding protein ZFP36[J]. Genome Biol, 2014, 15(1): R12. DOI:10.1186/gb2014151r12.
[16]MUSGROVE E A, CALDON C E, BARRACLOUGH J, et al. Cyclin D as a therapeutic target in cancer[J]. Nat Rev Cancer, 2011, 11(8): 558-572. DOI:10.1038/nrc3090.
[17]LEE H H, LEE S R, LEEM S H. Tristetraprolin regulates prostate cancer cell growth through suppression of E2F1[J]. J Microbiol Biotechnol, 2014, 24(2): 287-294. DOI:10.4014/jmb.1309.09070.
[18]LEE S R, ROH Y G, KIM S K, et al. Activation of EZH2 and SUZ12 regulated by E2F1 predicts the disease progression and aggressive characteristics of bladder cancer[J]. Clin Cancer Res, 2015, 21(23): 5391-5403. DOI:10.1158/10780432.CCR142680.
[19]ZHANG Y, WANG Z, MAGNUSON N S. Pim1 kinasedependent phosphorylation of p21Cip1/WAF1 regulates its stability and cellular localization in H1299 cells[J]. Mol Cancer Res, 2007, 5(9): 909-922. DOI:10.1158/15417786.MCR060388.
[20]ROUNBEHLER R J, FALLAHI M, YANG C, et al. Tristetraprolin impairs mycinduced lymphoma and abolishes the malignant state[J]. Cell, 2012, 150(3): 563-574. DOI:10.1016/j.cell.2012.06.033.
[21] SANDUJA S, KAZA V, DIXON D A. The mRNA decay factor tristetraprolin (TTP) induces senescence in human papillomavirustransformed cervical cancer cells by targeting E6AP ubiquitin ligase[J]. Aging (Albany NY), 2009, 1(9): 803-817. DOI:10.18632/aging.100086.
[22]GUO J, QU H, SHAN T, et al. Tristetraprolin overexpression in gastric cancer cells suppresses PDL1 expression and inhibits tumor progression by enhancing antitumor immunity[J]. Mol Cells, 2018, 41(7): 653-664. DOI:10.14348/molcells.2018.0040.
[23]TRAN D D H, KOCH A, ALLISTER A, et al. Treatment with MAPKAP2 (MK2) inhibitor and DNA methylation inhibitor, 5aza dC, synergistically triggers apoptosis in hepatocellular carcinoma (HCC) via tristetraprolin (TTP)[J]. Cell Signall, 2016, 28(12): 1872-1880. DOI:10.1016/j.cellsig.2016.09.002.
[24]DONG F, LI C, WANG P, et al. The RNA binding protein tristetraprolin downregulates autophagy in lung adenocarcinoma cells[J]. Exp Cell Res, 2018, 367(1): 89-96. DOI:10.1016/j.yexcr.2018.03.028.
[25]COELHO M A, DE CARN TRCESSON S, RANA S, et al. Oncogenic RAS signaling promotes tumor immunoresistance by stabilizing PDL1 mRNA[J]. Immunity, 2017, 47(6): 1083-1099.e6. DOI:10.1016/j.immuni.2017.11.016.

相似文献/References:

[1]刘翩,王斌,吴国明.吉非替尼维持治疗晚期肺腺癌长期生存1例[J].第三军医大学学报,2012,34(15):1491.
[2]刘瑞青,申淑景,马杰,等.肺腺癌组织中TTF-1和SPA的表达对EGFR突变的临床预测价值[J].第三军医大学学报,2012,34(22):2314.
 Liu Ruiqing,Shen Shujing,Ma Jie,et al.Clinical value of TTF-1 and SPA expression in predicting EGFR mutations in lung adenocarcinoma[J].J Third Mil Med Univ,2012,34(09):2314.
[3]刘小军,张雪琳,李莎莎,等.172例肺腺癌脑转移影响因素的临床分析[J].第三军医大学学报,2013,35(01):73.
 Liu Xiaojun,Zhang Xuelin,Li Shasha,et al.Clinical analysis of influencing factors for 172 lung adenocarcinoma patients with brain metastasis[J].J Third Mil Med Univ,2013,35(09):73.
[4]余祖滨,白莉,姚珂,等.SOCS3对肺腺癌细胞株A549迁移和侵袭能力的影响[J].第三军医大学学报,2008,30(07):588.
 YU Zu-bin,BAI Li,YAO Ke,et al.Effects of SOCS3 gene on migration and invasion of lung adenocarcinoma cell line A549[J].J Third Mil Med Univ,2008,30(09):588.
[5]朱冰,杨建茹,付新平,等.GALV致融性糖蛋白转导肺腺癌细胞的抗肿瘤实验研究[J].第三军医大学学报,2007,29(18):1737.
 ZHU Bing,YANG Jian-ru,FU Xin-ping,et al.Antitumor effect of GALV membrane fusion glycoprotein on lung adenocarcinoma in vivo and in vitro[J].J Third Mil Med Univ,2007,29(09):1737.
[6]庞华,罗弋,李少林.人源肺腺癌噬菌体抗体库的构建及初步筛选[J].第三军医大学学报,2008,30(22):2082.
 PANG Hua,LUO Yi,LI Shao-lin.Construction of and screening for phage antibody library of human lung adenocarcinoma[J].J Third Mil Med Univ,2008,30(09):2082.
[7]白莉,祝蓉,陈智鸿,等.靶向EGFR基因的siRNA表达载体构建及其生物学效应的研究[J].第三军医大学学报,2005,27(23):2307.
[8]熊玮,钱桂生,黄桂君.氯氨顺铂诱导耐药后肺腺癌细胞株生物学特性及染色体核型分析[J].第三军医大学学报,2005,27(22):2226.
[9]郭芮伶,吴国明,戢福云,等.多西紫杉醇对肺腺癌多药耐药细胞A549/CDDP及其亲代细胞作用机制的初步研究[J].第三军医大学学报,2005,27(07):610.
[10]张雪淋,原薇薇,傅华,等.HOXA10在肺腺癌中的过表达及其临床意义[J].第三军医大学学报,2013,35(05):468.
 Zhang Xuelin,Yuan Weiwei,Fu Hua,et al.Over-expression and clinical significance of HOXA10 in human lung adenocarcinoma[J].J Third Mil Med Univ,2013,35(09):468.

更新日期/Last Update: 2019-05-08