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 (2): 151-156.doi: 10.12007/j.issn.0258-4646.2018.02.013

• 论著 • 上一篇    下一篇

Hedgehog/Gli和PI3K/AKT信号通路的串话促进胃癌AZ521细胞上皮-间质转化

王雷1, 杜媛鲲2, 米源1, 廖海江1, 陈阁1, 张耀中1, 刘庆熠1   

  1. 1. 河北医科大学第四医院胸二科, 石家庄 050011;
    2. 河北医科大学期刊社, 石家庄 050017
  • 收稿日期:2017-07-11 出版日期:2018-02-28 发布日期:2018-01-11
  • 通讯作者: 刘庆熠 E-mail:qingyiliu@163.com
  • 作者简介:王雷(1976-),男,主任医师,博士后.
  • 基金资助:
    河北省科技厅科技攻关计划(132077127D);河北省卫计委医学重点科研基金(20160177)

Crosstalk of Hedgehog/Gli and PI3k/Akt Pathway Promotes Epithelial-mesenchymal Transition in Gastric Adenocarcinoma

WANG Lei1, DU Yuankun2, MI Yuan1, LIAO Haijiang1, CHEN Ge1, ZHANG Yaozhong1, LIU Qingyi1   

  1. 1. Department of Thoracic Surgery, The Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, China;
    2. Department of Periodical Magazine, Hebei Medical University, Shijiazhuang 050017, China
  • Received:2017-07-11 Online:2018-02-28 Published:2018-01-11

摘要: 目的 探讨Hedgehog/Gli促进胃癌AZ521细胞上皮-间质转化(EMT)的肿瘤分子学机制。方法 GANT61处理AZ521细胞24 h后,用实时荧光定量PCR法观察GANT61对Gli1Gli2N-cadherinE-cadherin mRNA表达的影响;用Western blotting观察GANT61对Gli1、Gli2、p-AKT、AKT、N-cadherin和E-cadherin蛋白表达的影响;用侵袭实验观察GANT61对胃癌细胞迁移侵袭能力的影响;同时,应用N-Shh刺激Hedgehog通路观察相应指标的变化。结果 与对照组相比,GANT61组胃癌AZ521细胞的Gli1Gli2N-cadherin mRNA表达显著下调,E-cadherin mRNA表达上调;Western blotting结果显示GANT61可以下调AZ521的Gli1、Gli2、p-AKT、N-cadherin蛋白表达,上调E-cadherin蛋白表达。侵袭实验结果显示GANT61可以显著抑制AZ521细胞的侵袭能力。应用N-Shh可以显著上调Gli1Gli2N-cadherin mRNA和蛋白表达以及p-AKT蛋白表达,下调E-cadherin mRNA及蛋白表达,促进肿瘤细胞侵袭转移。结论 Gli1和Gli2表达下调可以抑制胃癌细胞的侵袭转移,可能与Gli通过PI3K/AKT途径促进EMT有关。

关键词: 胃癌, Hedgehog/Gli, PI3K/AKT, GANT61, 上皮-间质转化

Abstract: Objective To investigate the tumor molecular mechanism of Hedgehog/Gli in promoting the epithelial-mesenchymal transition (EMT) in gastric cancer AZ521 cells.Methods After 24 h of treatment with GANT61,the mRNA expression of Gli1,Gli2,N-cadherin,and E-cadherin in the AZ521 cell line were detected by real-time fluorescence quantitative PCR. A Western blotting assay was conducted to determine the expression of the above cytokines,p-AKT and AKT. The effect of GANT61 on invasion was observed by transwell assay. N-Shh stimulation of the Hedgehog pathway was conducted to confirm the changes in these cytokines.Results GANT61 significantly downregulated the mRNA expression of Gli1,Gli2,and N-cadherin,but upregulated E-cadherin mRNA expression. The Western blotting assay revealed that GANT61 downregulated the protein expression of Gli1,Gli2,p-AKT,and N-cadherin,but upregulated E-cadherin expression. Furthermore,GANT61 inhibited the invasion. N-Shh proteins up-regulated Gli1,Gli2,and N-cadherin mRNA,protein expression and p-AKT protein expression,but downregulated E-cadherin mRNA and protein expressions. N-Shh promoted the invasion of tumor cells.Conclusion Downregulation of Gli1 and Gli2 can inhibit the invasion and metastasis in gastric cancer cells,which may be related to the promotion of EMT by Gli through the PI3K/AKT pathway.

Key words: gastric adenocarcinoma, Hedgehog/Gli, PI3K/AKT, GANT61, epithelial-mesenchymal transition

中图分类号: 

  • R73
[1] JEMAL A,BRAY F,CENTER MM,et al. Global cancer statistics[J]. CA Cancer J Clin,2011,61(2):69-90. DOI:10.3322/caac.20107.
[2] LI H,YUE D,JIN JQ,et al. Gli promotes epithelial-mesenchymal transition in human lung adenocarcinomas[J]. Oncotarget,2016,7(49):80415-80425. DOI:10.18632/oncotarget.11246.
[3] SHIMO T,MATSUMOTO K,TAKABATAKE K,et al. The role of sonic hedgehog signaling in osteoclastogenesis and jaw bone destruction[J]. PLoS One,2016,11(3):e0151731. DOI:10.1371/journal.pone.0151731.
[4] SONG L,WANG W,LIU D,et al. Targeting of sonic hedgehog-Gli signaling:a potential therapeutic target for patients with breast cancer[J]. Oncol Lett,2016,12(2):1027-1033. DOI:10.3892/ol.2016.4722.
[5] YAN Z,XU L,ZHANG J,et al. Aldehyde dehydrogenase 1A1 stabilizes transcription factor Gli2 and enhances the activity of Hedgehog signaling in hepatocellular cancer[J]. Biochem Biophys Res Commun,2016,471(4):466-473. DOI:10.1016/j.bbrc.2016.02.052.
[6] TAIPALE J,BEACHY PA. The Hedgehog and Wnt signalling pathways in cancer[J]. Nature,2001,411(6835):349-354.
[7] STECCA B,RUIZ IAA. Context-dependent regulation of the GLI code in cancer by HEDGEHOG and non-HEDGEHOG signals[J]. J Mol Cell Biol,2010,2(2):84-95. DOI:10.1093/jmcb/mjp052.
[8] AASRUM M,TJOMSLAND V,THORESENJ GH,et al. PI3K is required for both basal and LPA-induced DNA synthesis in oral carcinoma cells[J]. J Oral Pathol Med,2016,45(6):425-432. DOI:10.1111/jop.12384.
[9] FRASSON C,RAMPAZZO E,ACCORDI B,et al. Inhibition of PI3K signalling selectively affects medulloblastoma cancer stem cells[J]. Biomed Res Int,2015,2015:973912. DOI:10.1155/2015/973912.
[10] ABERGER F,KERN D,GRIL R,et al. Canonical and noncanonical Hedgehog/GLI signaling in hematological malignancies[J]. Vitam Horm,2012,88:25-54. DOI:10.1016/B978-0-12-394622-5.00002-X.
[11] DORMORY V,DANILIN S,LINDNER V,et al. The sonic hedgehog signaling pathway is reactivated in human renal cell carcinoma and plays orchestral role in tumor growth[J]. Mol Cancer,2009,8:123. DOI:10.1186/1476-4598-8-123.
[12] KERN D,REGL G,HOFBAUER SW,et al. Hedgehog/GLI and PI3K signaling in the initiation and maintenance of chronic lymphocytic leukemia[J]. Oncogene,2015,34(42):5341-5351. DOI:10.1038/onc.2014.450.
[13] GONNISSEN A,ISEBAEERT S,HAUSTERTERMANS K. Targeting the Hedgehog signaling pathway in cancer:beyond smoothened[J]. Oncotarget,2015,6(16):13899-13913. DOI:10.18632/oncotarget.4224.
[14] ZHOU J,ZHU G,HUANG J,et al. Non-canonical GLI1/2 activation by PI3K/AKT signaling in renal cell carcinoma:a novel potential therapeutic target[J]. Cancer Lett,2016,370(2):313-323. DOI:10.1016/j.canlet.2015.11.006.
[15] MAJUMDER PK,SELLERS WR. Akt-regulated pathways in prostate cancer[J]. Oncogene,2005,24(50):7465-7474. DOI:10.1038/sj.onc.1209096.
[16] AMIN H,WANI NA,FAROOQ S,et al. Inhibition of invasion in pancreatic cancer cells by conjugate of EPA with beta (3,3) -Pip-OH via PI3K/Akt/NF-kB pathway[J]. ACS Med Chem Lett,2015,6(10):1071-1074. DOI:10.1021/acsmedchemlett.5b00257.
[17] ABRAHAM J. PI3K/AKT/mTOR pathway inhibitors:the ideal combination partners for breast cancer therapies?[J]. Expert Rev Anticancer Ther,2015,15(1):51-68. DOI:10.1586/14737140.2015. 961429.
[18] SUNG WJ,PARK KS,KWAK SG,et al. Epithelial-mesenchymal transition in patients of pulmonary adenocarcinoma:correlation with cancer stem cell markers and prognosis[J]. Int J Clin Exp Pathol,2015,8(8):8997-9009.
[19] YAN X,YAN L,LIU S,et al. N-cadherin,a novel prognostic biomarker,drives malignant progression of colorectal cancer[J]. Mol Med Rep,2015,12(2):2999-3006. DOI:10.3892/mmr.2015.3687.
[20] ATTRAMADAL CG,KUMAR S,BOYSEN ME,et al. Tumor budding,emt and cancer stem cells in T1-2/N0 oral squamous cell carcinomas[J]. Anticancer Res,2015,35(11):6111-6120.
[21] PATIL PU,D'AMBROSIO J,INGE LJ,et al. Carcinoma cells induce lumen filling and EMT in epithelial cells through soluble E-cadherin-mediated activation of EGFR[J]. J Cell Sci,2015,128(23):4366-4379. DOI:10.1242/jcs.173518.
[22] ISLAM SS,MOKHTARI RB,NOMAN AS,et al. Sonic hedgehog (Shh) signaling promotes tumorigenicity and stemness via activation of epithelial-to-mesenchymal transition (EMT) in bladder cancer[J]. Mol Carcinog,2016,55(5):537-551. DOI:10.1002/mc.22300.
[23] FU J,RODOVA M,ROY SK,et al. GANT-61 inhibits pancreatic cancer stem cell growth in vitro and in NOD/SCID/IL2R gamma null mice xenograft[J]. Cancer Lett,2013,330(1):22-32. DOI:10.1016/j.canlet.2012.11.018.
[24] SRIVASTAVA RK,KAYLANI SZ,EDRESS N,et al. GLI inhibitor GANT-61 diminishes embryonal and alveolar rhabdomyosarcoma growth by inhibiting Shh/AKT-mTOR axis[J]. Oncotarget,2014,5(23):12151-12165. DOI:10.18632/oncotarget.2569.
[25] LIN Z,LI S,SHENG H,et al. Suppression of GLI sensitizes medulloblastoma cells to mitochondria-mediated apoptosis[J]. J Cancer Res Clin Oncol,2016,142(12):2469-2478. DOI:10.1007/s00432-016-2241-1.
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