中国医科大学学报

中国医科大学学报
  • 中文核心期刊
  • 中国科技核心期刊
  • 中国高校百佳科技期刊
  • BA、CA收录

中国医科大学学报 ›› 2018, Vol. 47 ›› Issue (4): 321-328.doi: 10.12007/j.issn.0258-4646.2018.04.008

• 论著 • 上一篇    下一篇

SIRT1沉默对人甲状腺乳头状癌细胞株TPC-1生物学功能的影响

彭华童1, 吴文艺1, 张丽婷2, 林建清1, 余艺煌1, 吴春林1, 黄种心1, 邱建龙3   

  1. 1. 福建医科大学附属第二医院肿瘤外科, 福建 泉州 362000;
    2. 解放军第一八零医院内分泌科, 福建 泉州 362000;
    3. 解放军第一八零医院病理科, 福建 泉州 362000
  • 收稿日期:2017-09-28 出版日期:2018-04-30 发布日期:2018-04-10
  • 通讯作者: 吴文艺 E-mail:wwyii8522858@163.com
  • 作者简介:彭华童(1990-),男,硕士研究生.
  • 基金资助:
    福建省卫生系统中青年骨干人才培养项目(2015-ZQNZD-26)

Effect of Silencing SIRT1 on the Biological Behavior of the Human Thyroid Papillary Carcinoma Cell Line TPC-1

PENG Huatong1, WU Wenyi1, ZHANG Liting2, LIN Jianqing1, YU Yihuang1, WU Chunlin1, HUANG Zhongxin1, QIU Jianlong3   

  1. 1. Department of Oncological Surgery, The Second Affiliated Hospital of Fujian Medical University, Quanzhou 362000, China;
    2. Department of Endocrinology, The 180 th Military Hospital of Chinese People's Liberation Army, Quanzhou 362000, China;
    3. Department of Pathology, The 180 th Military Hospital of Chinese People's Liberation Army, Quanzhou 362000, China
  • Received:2017-09-28 Online:2018-04-30 Published:2018-04-10

摘要: 目的 观察沉默信息调节因子2相关酶1(SIRT1)基因沉默对人甲状腺乳头状癌细胞株TPC-1生物学功能的影响。方法 利用SIRT1siRNA技术下调SIRT1在甲状腺乳头状癌细胞株TPC-1中的表达,应用qRT-PCR方法检测siRNA的干扰效率,筛选出干扰效率最强的siRNA,并用Western blotting验证SIRT1蛋白的表达,利用CCK8法、β-半乳糖苷酶染色法、Hoechst染色法及流式细胞技术检测转染细胞的增殖抑制率、衰老、凋亡与周期的影响。结果 转染SIRT1siRNA的甲状腺乳头状癌细胞较未转染细胞SIRT1 mRNA及蛋白表达水平显著降低(P < 0.05);转染细胞增殖率显著低于未转染细胞(P < 0.05),转染细胞增殖明显受到抑制(P < 0.05);转染组衰老细胞比例明显增加(P < 0.05),凋亡细胞比例明显升高(P < 0.05);转染细胞出现G1期停滞(67.99%±0.87%,P < 0.05)。结论 SIRT1在人甲状腺乳头状癌细胞株TPC-1的低表达能抑制细胞的增殖能力,阻滞细胞周期,并诱导细胞的衰老及凋亡。

关键词: 沉默信息调节因子2相关酶1, RNA干扰, 甲状腺乳头状癌

Abstract: Objective To investigate the effect of silencing SIRT1 on the biological behavior of the human thyroid papillary carcinoma cell line TPC-1. Methods SIRT1 siRNA was used to downregulate the expression of SIRT1 in TPC-1. The interference efficiency of siRNA was detected using qRT-PCR. The siRNA with the strongest interference efficiency was selected and the expression of SIRT1 protein was verified by Western blotting. The inhibitory rate of proliferation,senescence,apoptosis,and cycling of transfected cells was detected using the CCK8 method,β-galactosidase staining,Hoechst staining,and flow cytometry,respectively. Results The SIRT1 expression in thyroid papillary carcinoma cells transfected with siRNA was lower (P < 0.05) than that in the untransfected cells. The proliferation of transfected cells was significantly inhibited (P < 0.05) and their proliferation rate was lower than that of untransfected cells (P < 0.05). The proportion of aged cells and the percentage of apoptotic cells in the transfected carcinoma cells was significantly increased (P < 0.05). The transfection of SIRT1 siRNA increased the number of cells in the G1 phase (67.99%±0.87%,P < 0.05). Conclusion Reduced SIRT1 expression in the human thyroid papillary carcinoma cell line TPC-1 can inhibit cell proliferation,block the cell cycle,and induce cell senescence and apoptosis.

Key words: SIRT1, RNA interference, thyroid papillary carcinoma

中图分类号: 

  • R653
[1] LIVOLSI VA. Papillary thyroid carcinoma:an update[J]. Mod Pathol, 2011,24:S1-S9. DOI:10.1038/modpathol.2010.129.
[2] HATTORI Y,IHARA M. SIRT1[J]. Nihon Rinsho,2016,74(4):589-594.
[3] VIKRAM A,LEWARCHIK CM,YOON JY,et al. Sirtuin 1 regulates cardiac electrical activity by deacetylating the cardiac sodium channel[J]. Nat Med,2017,23(3):361-367. DOI:10.1038/nm.4284.
[4] HERRANZ D,MARAVER A,CANAMERO M,et al. SIRT1 promotes thyroid carcinogenesis driven by PTEN deficiency[J]. Oncogene, 2013,32(34):4052-4056. DOI:10.1038/onc.2012.407.
[5] 吴文艺,张丽婷,傅德强,等. 沉默信息调节因子2相关酶1在甲状腺乳头状癌组织中的表达及其意义[J]. 中华实验外科杂志, 2015,32(4):877-880. DOI:10.3760/cma.j.issn.1001-9030.2015.04.072.
[6] WU W,ZHANG L,LIN J,et al. Hypermethylation of the HIC1 promoter and aberrant expression of HIC1/SIRT1 contribute to the development of thyroid papillary carcinoma[J]. Oncotarget,2016,7(51):84416-84427. DOI:10.18632/oncotarget.12936.
[7] HAIGIS MC,GUARENTE LP. Mammalian sirtuins——emerging roles in physiology,aging,and calorie restriction[J]. Genes Dev,2006,20(21):2913-2921. DOI:10.1101/gad.1467506.
[8] MIN J,LANDRY J,STERNGLANZ R,et al. Crystal structure of a sir2 homolog-nad complex[J]. Cell,2001,105(2):269-279. DOI:10.1016/S0092-8674(01) 00317-8.
[9] AQUILANO K,VIGILANZA P,BALDELLI S,et al. Peroxisome proliferator-activated receptor gamma co-activator 1alpha (pgc-1alpha) and sirtuin 1(sirt1) reside in mitochondria:possible direct function in mitochondrial biogenesis[J]. J Biol Chem,2010,285(28):21590-21599. DOI:10.1074/jbc.M109.070169.
[10] BENDALE DS,KARPE PA,CHHABRA R,et al. 17-beta Oestradiol prevents cardiovascular dysfunction in post-menopausal metabolic syndrome by affecting SIRT1/AMPK/H3 acetylation[J]. Br J Phar macol,2013,170(4):779-795. DOI:10.1111/bph.12290.
[11] LI L,WANG L,LI L,et al. Activation of p53 by sirt1 inhibition enhances elimination of cml leukemia stem cells in combination with imatinib[J]. Cancer Cell,2012,21(2):266-281. DOI:10.1016/j.ccr.2011.12.020.
[12] GONFLONI S,IANNIZZOTTO V,MAIANI E,et al. P53 and Sirt1:routes of metabolism and genome stability[J]. Biochem Pharmacol, 2014,92(1):149-156. DOI:10.1016/j.bcp.2014.08.034.
[13] ZHOU N,LIN X,DONG W,et al. SIRT1 alleviates senescence of degenerative human intervertebral disc cartilage endo-plate cells via the p53/p21 pathway[J]. Sci Rep,2016,6:22628. DOI:10.1038/srep22628.
[14] HORI YS,KUNO A,HOSODA R,et al. Regulation of FOXOs and p53 by SIRT1 modulators under oxidative stress[J]. PLoS One, 2013,8(9):e73875. DOI:10.1371/journal.pone.0073875.
[15] LEE D,GOLDBERG AL. SIRT1 protein,by blocking the activities of transcription factors FoxO1 and FoxO3,inhibits muscle atrophy and promotes muscle growth[J]. J Biol Chem,2013,288(42):30515-30526. DOI:10.1074/jbc.M113.489716.
[16] GU X,HAN D,CHEN W,et al. SIRT1-mediated FoxOs pathways protect against apoptosis by promoting autophagy in osteoblast-like MC3T3-E1 cells exposed to sodium fluoride[J]. Oncotarget,2016,7(40):65218-65230. DOI:10.18632/oncotarget.11573.
[17] KIM MJ,HONG KS,KIM HB,et al. Ku70 acetylation and modulation of c-Myc/ATF4/CHOP signaling axis by SIRT1 inhibition lead to sensitization of HepG2 cells to TRAIL through induction of DR5 and down-regulation of c-FLIP[J]. Int J Biochem Cell Biol,2013,45(3):711-723. DOI:10.1016/j.biocel.2012.12.005.
[18] ROTH M,WANG Z,CHEN WY. SIRT1 and LSD1 competitively regulate KU70 functions in DNA repair and mutation acquisition in cancer cells[J]. Oncotarget,2016,7(31):50195-50214. DOI:10.18632/oncotarget.10328.
[19] KUNO A,HORI YS,HOSODA R,et al. Resveratrol improves cardiomyopathy in dystrophin-deficient mice through SIRT1 protein-mediated modulation of p300 protein[J]. J Biol Chem,2013,288(8):5963-5672. DOI:10.1074/jbc.M112.392050.
[20] ZENG Z,CHENG S,CHEN H,et al. Activation and overexpression of Sirt1 attenuates lung fibrosis via P300[J]. Biochem Biophys Res Commun,2017,486(4):1021-1026. DOI:10.1016/j.bbrc.2017.03.155.
[21] CURTIL C,ENACHE LS,RADREAU P,et al. The metabolic sensors FXRalpha,PGC-1alpha,and SIRT1 cooperatively regulate hepatitis B virus transcription[J]. FASEB J,2014,28(3):1454-1463. DOI:10.1096/fj.13-236372.
[22] TRABOULSI H,DAVOLI S,CATEZ P,et al. Dynamic partnership between TFⅡH,PGC-1alpha and SIRT1 is impaired in trichothiodystrophy[J]. PLoS Genet,2014,10(10):e1004732. DOI:10.1371/journal.pgen.1004732.
[23] KAUPPINEN A,SUURONEN T,OJALA J,et al. Antagonistic crosstalk between NF-kappaB and SIRT1 in the regulation of inflammation and metabolic disorders[J]. Cell Signal,2013,25(10):1939-1948. DOI:10.1016/j.cellsig.2013.06.007.
[24] CHEN Y,LIU H,ZHANG H,et al. The sirt1/NF-kB signaling pathway is involved in regulation of endothelin type B receptors mediated by homocysteine in vascular smooth muscle cells[J]. Biomed Pharmacother,2016,84:1979-1985. DOI:10.1016/j.biopha.2016.11.011.
[25] 邓向群. SIRT1表达对大鼠胰岛β细胞凋亡及非酒精性脂肪肝病形成的影响[D].华中科技大学,2007.
[26] HOLLOWAY KR,BARBIERI A,MALYARCHUK S,et al. SIRT1 positively regulates breast cancer associated human aromatase (CYP19A1) expression[J]. Mol Endocrinol,2013,27(3):480-490. DOI:10.1210/me.2012-1347.
[27] NOH SJ,BAEK HA,PARK HS,et al. Expression of SIRT1 and cortactin is associated with progression of non-small cell lung cancer[J]. Pathol Res Pract,2013,209(6):365-370. DOI:10.1016/j.prp.2013.03.011.
[28] KRIEGL L,VIETH M,KIRCHNER T,et al. Up-regulation of c-myc and sirt1 expression correlates with malignant transformation in the serrated route to colorectal cancer[J]. Oncotarget,2012,3(10):1182-1193. DOI:10.18632/oncotarget.628.
[29] SIMIC P,WILLIAMS EO,BELL EL,et al. SIRT1 suppresses the epithelial-to-mesenchymal transition in cancer metastasis and organ fibrosis[J]. Cell Rep,2013,3(4):1175-1186. DOI:10.1016/j.celrep.2013.03.019.
[30] SUN L,LI H,CHEN J,et al. A SUMOylation-dependent pathway regulates SIRT1 transcription and lung cancer metastasis[J]. J Natl Cancer Inst,2013,105(12):887-898. DOI:10.1093/jnci/djt118.
[1] 杜媛鲲, 赵永波, 米源, 陈阁, 廖海江, 王雷. 小干扰RNA下调PLK1表达对食管腺癌细胞肿瘤生物学行为的影响[J]. 中国医科大学学报, 2018, 47(10): 905-908,913.
[2] 齐晓莹,蒋中秀,刘洋,李宁,张晓晔. 沉默信息调节因子2相关酶1在直肠癌中的表达及临床意义[J]. 中国医科大学学报, 2015, 44(7): 606-610.
[3] 林新恭,吴文艺,张丽婷,黄种心,王朝阳. HIC1和SIRT1的异常表达与甲状腺乳头状癌的关系研究[J]. 中国医科大学学报, 2015, 44(11): 995-998.
[4] 贾爱庆,崔俊帅,董文武,王志宏,贺亮,张大林,吕承洲,张浩. HSP90和Sufu在甲状腺乳头状癌中的表达及意义[J]. 中国医科大学学报, 2014, 43(3): 231-234.
[5] 宫琦玉,汲坤,张丽艳,王波,楚琪,张明璇. RNA干扰抑制乳腺癌MCF?7细胞鼠双微体?2基因表达及细胞增殖的实验研究[J]. 中国医科大学学报, 2014, 43(10): 892-896.
[6] 孙倩,孙磊,金镇,范杰慧,息小雪. RNA干扰技术抑制SHBG基因表达对滋养细胞凋亡的影响[J]. 中国医科大学学报, 2013, 42(8): 689-692.
[7] 祁馨卉,崔慧霞,马楠,姜又红. GPC3基因重组慢病毒载体的构建及其在肝癌细胞中的表达[J]. 中国医科大学学报, 2013, 42(8): 726-729.
[8] 吴文艺,张丽婷,王朝阳,邱建龙,朱世泽. 叉头框转录因子M1在甲状腺乳头状癌组织中的表达及意义[J]. 中国医科大学学报, 2013, 42(2): 127-130.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!

中国医科大学学报版权所有©2018

未经允许,严禁擅自转载本站图文资料

地址:中国 沈阳市沈北新区蒲河路77号 110122

辽ICP备05014850

JOURNAL OF CHINA MEDICAL UNIVERSITY

ADDRESS: NO.77 PUHE ROAD

SHENYANG NORTH NEW AREA, SHENYANG

LIAONING PROVINCE, P.R. CHINA