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Notch1 and PI3K/Akt signaling blockers DAPT and LY294002 coordinately inhibit metastasis of gastric cancer through mutual enhancement

  • Xingchun Peng
  • Jianrui Zhou
  • Bin Li
  • Tao Zhang
  • Yamin Zuo
  • Xinsheng GuEmail author
Original Article
  • 86 Downloads

Abstract

Purpose

Blockade of either Notch1 or PI3K/Akt pathway inhibits metastasis of gastric cancer. However, whether blockade of both pathways coordinately exerts such an effect remains unknown. In this study, we aimed to investigate the effects of combined treatment with Notch1 signaling blocker DAPT and PI3K/Akt signal blocker LY294002 on metastasis of gastric cancer.

Methods

Notch intracellular domain (NICD) and phosphorylated Akt (p-Akt) levels in gastric cancer tissues and their adjacent normal tissue samples and gastric cancer SGC7901 and AGS cells and normal GES-1 cells were determined using immunohistochemistry and Western blotting. The effects of combined DAPT and LY294002 on metastasis of gastric cancer were evaluated by examining migration and invasion potential of SGC7901 cells using wound healing and transwell assays, determining changes in the levels of epithelial–mesenchymal transition biomarkers and MMP-9, Notch1, HES1, and phosphorylation of Akt in gastric cancer SGC7901 cells and/or AGS cells in vitro using Western blotting, and metastasis of gastric cancer to lungs in BALB/c nude mice after treatment.

Results

NICD and p-Akt levels were significantly higher in gastric cancer tissues and SGC7901 and AGS cells than those in the normal control and GES-1 cells. Migration and invasion potential of SGC7901 cells, EMT biomarkers and MMP-9 in SGC7901 cells, and metastasis of gastric cancer to lungs in mice were coordinately inhibited by DAPT and LY294002. In addition, DAPT and LY294002 coordinately inhibited the levels of Notch1, HES1, and p-Akt in gastric cancer cells.

Conclusion

DAPT and LY294002 coordinately inhibited metastasis of gastric cancer through mutual enhancement.

Keywords

Gastric cancer Notch1 PI3K/Akt Metastasis 

Notes

Author contributions

XP, JZ, and BL collected and analyzed data. TZ and YZ collected data. XP and XG conceived the study and wrote the manuscript.

Funding

This work was supported by the Department of Education of Hubei Province (Grant No. B2018109), Natural Science Foundation of Hubei Province of China (2016CFB530), Faculty Development Foundation of Hubei University of Medicine (2014QDJZR01), and Department of Health of Xuhui District of Shanghai (SHXH201402).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. 1.
    Schlesinger-Raab A, Mihaljevic AL, Egert S, Emeny R, Jauch KW, Kleeff J, Novotny A, Nussler NC, Rottmann M, Schepp W, Schmitt W, Schubert-Fritschle G, Weber B, Schuhmacher C, Engel J (2016) Outcome of gastric cancer in the elderly: a population-based evaluation of the Munich Cancer Registry. Gastric Cancer 19(3):713–722.  https://doi.org/10.1007/s10120-015-0527-7 CrossRefPubMedGoogle Scholar
  2. 2.
    Tekesin K, Emin Gunes M, Tural D, Akar E, Zirtiloglu A, Karaca M, Selcukbiricik F, Bayrak S, Ozet A (2019) Clinicopathological characteristics, prognosis and survival outcome of gastric cancer in young patients: a large cohort retrospective study. J BUON 24(2):672–678PubMedGoogle Scholar
  3. 3.
    Yang SW, Ping YF, Jiang YX, Luo X, Zhang X, Bian XW, Yu PW (2016) ATG4A promotes tumor metastasis by inducing the epithelial–mesenchymal transition and stem-like properties in gastric cells. Oncotarget 7(26):39279–39292.  https://doi.org/10.18632/oncotarget.9827 CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Yang K, Choi YY, Zhang WH, Chen XZ, Song MK, Lee J, Zhang B, Chen ZX, Kim HI, Chen JP, Cheong JH, Zhou ZG, Hyung WJ, Hu JK, Noh SH (2016) Strategies to improve treatment outcome in gastric cancer: a retrospective analysis of patients from two high-volume hospitals in Korea and China. Oncotarget 7(28):44660–44675.  https://doi.org/10.18632/oncotarget.9378 CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Gramaticu IM, Croitoru AE (2018) Current management of gastric cancer in Europe. Chirurgia (Bucur) 113(6):758–764.  https://doi.org/10.21614/chirurgia.113.6.758 CrossRefGoogle Scholar
  6. 6.
    Tokunaga M, Makuuchi R, Miki Y, Tanizawa Y, Bando E, Kawamura T, Terashima M (2016) Surgical and survival outcome following truly palliative gastrectomy in patients with incurable gastric cancer. World J Surg 40(5):1172–1177.  https://doi.org/10.1007/s00268-015-3398-5 CrossRefPubMedGoogle Scholar
  7. 7.
    Qi X, Liu Y, Wang W, Cai D, Li W, Hui J, Liu C, Zhao Y, Li G (2016) Management of advanced gastric cancer: an overview of major findings from meta-analysis. Oncotarget 7(47):78180–78205.  https://doi.org/10.18632/oncotarget.12102 CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Russo AE, Strong VE (2019) Gastric cancer etiology and management in Asia and the West. Annu Rev Med 70:353–367.  https://doi.org/10.1146/annurev-med-081117-043436 CrossRefPubMedGoogle Scholar
  9. 9.
    Henrique D, Schweisguth F (2019) Mechanisms of Notch signaling: a simple logic deployed in time and space. Development.  https://doi.org/10.1242/dev.172148 CrossRefPubMedGoogle Scholar
  10. 10.
    Siebel C, Lendahl U (2017) Notch signaling in development, tissue homeostasis, and disease. Physiol Rev 97(4):1235–1294.  https://doi.org/10.1152/physrev.00005.2017 CrossRefPubMedGoogle Scholar
  11. 11.
    Kovall RA, Gebelein B, Sprinzak D, Kopan R (2017) The Canonical notch signaling pathway: structural and biochemical insights into shape, sugar, and force. Dev Cell 41(3):228–241.  https://doi.org/10.1016/j.devcel.2017.04.001 CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Du X, Cheng Z, Wang YH, Guo ZH, Zhang SQ, Hu JK, Zhou ZG (2014) Role of Notch signaling pathway in gastric cancer: a meta-analysis of the literature. World J Gastroenterol 20(27):9191–9199.  https://doi.org/10.3748/wjg.v20.i27.9191 CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Li W, Wang D, Sun X, Zhang Y, Wang L, Suo J (2019) ADAM17 promotes lymph node metastasis in gastric cancer via activation of the Notch and Wnt signaling pathways. Int J Mol Med 43(2):914–926.  https://doi.org/10.3892/ijmm.2018.4028 CrossRefPubMedGoogle Scholar
  14. 14.
    Ren Z, Zhang C, Ma L, Zhang X, Shi S, Tang D, Xu J, Hu Y, Wang B, Zhang F, Zheng H (2019) Lysophosphatidic acid induces the migration and invasion of SGC-7901 gastric cancer cells through the LPA2 and Notch signaling pathways. Int J Mol Med.  https://doi.org/10.3892/ijmm.2019.4186 CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Miao ZF, Xu H, Xu HM, Wang ZN, Zhao TT, Song YX, Xu YY (2017) DLL4 overexpression increases gastric cancer stem/progenitor cell self-renewal ability and correlates with poor clinical outcome via Notch-1 signaling pathway activation. Cancer Med 6(1):245–257.  https://doi.org/10.1002/cam4.962 CrossRefPubMedGoogle Scholar
  16. 16.
    Hennessy BT, Smith DL, Ram PT, Lu Y, Mills GB (2005) Exploiting the PI3K/Akt pathway for cancer drug discovery. Nat Rev Drug Discov 4(12):988–1004.  https://doi.org/10.1038/nrd1902 CrossRefPubMedGoogle Scholar
  17. 17.
    Yudushkin I (2019) Getting the Akt together: guiding intracellular Akt activity by PI3K. Biomolecules.  https://doi.org/10.3390/biom9020067 CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Noorolyai S, Shajari N, Baghbani E, Sadreddini S, Baradaran B (2019) The relation between PI3K/Akt signalling pathway and cancer. Gene 698:120–128.  https://doi.org/10.1016/j.gene.2019.02.076 CrossRefPubMedGoogle Scholar
  19. 19.
    Hao NB, Tang B, Wang GZ, Xie R, Hu CJ, Wang SM, Wu YY, Liu E, Xie X, Yang SM (2015) Hepatocyte growth factor (HGF) upregulates heparanase expression via the PI3K/Akt/NF-kappaB signaling pathway for gastric cancer metastasis. Cancer Lett 361(1):57–66.  https://doi.org/10.1016/j.canlet.2015.02.043 CrossRefPubMedGoogle Scholar
  20. 20.
    Zhang Y, Li Z, Fan X, Xiong J, Zhang G, Luo X, Li K, Jie Z, Cao Y, Huang Z, Wu F, Xiao L, Duan G, Chen H (2018) PRL-3 promotes gastric cancer peritoneal metastasis via the PI3K/Akt signaling pathway in vitro and in vivo. Oncol Lett 15(6):9069–9074.  https://doi.org/10.3892/ol.2018.8467 CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Liu JY, Jiang L, He T, Liu JJ, Fan JY, Xu XH, Tang B, Shi Y, Zhao YL, Qian F, Wang Y, Cui YH, Yu PW (2019) NETO2 promotes invasion and metastasis of gastric cancer cells via activation of PI3K/Akt/NF-kappaB/Snail axis and predicts outcome of the patients. Cell Death Dis 10(3):162.  https://doi.org/10.1038/s41419-019-1388-5 CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Ye B, Jiang LL, Xu HT, Zhou DW, Li ZS (2012) Expression of PI3K/Akt pathway in gastric cancer and its blockade suppresses tumor growth and metastasis. Int J Immunopathol Pharmacol 25(3):627–636.  https://doi.org/10.1177/039463201202500309 CrossRefPubMedGoogle Scholar
  23. 23.
    Xing X, Zhang L, Wen X, Wang X, Cheng X, Du H, Hu Y, Li L, Dong B, Li Z, Ji J (2014) PP242 suppresses cell proliferation, metastasis, and angiogenesis of gastric cancer through inhibition of the PI3K/Akt/mTOR pathway. Anticancer Drugs 25(10):1129–1140.  https://doi.org/10.1097/CAD.0000000000000148 CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Villegas SN, Gombos R, Garcia-Lopez L, Gutierrez-Perez I, Garcia-Castillo J, Vallejo DM, Da Ros VG, Ballesta-Illan E, Mihaly J, Dominguez M (2018) PI3K/Akt cooperates with oncogenic notch by inducing nitric oxide-dependent inflammation. Cell Rep 22(10):2541–2549.  https://doi.org/10.1016/j.celrep.2018.02.049 CrossRefPubMedGoogle Scholar
  25. 25.
    Graziani I, Eliasz S, De Marco MA, Chen Y, Pass HI, De May RM, Strack PR, Miele L, Bocchetta M (2008) Opposite effects of Notch-1 and Notch-2 on mesothelioma cell survival under hypoxia are exerted through the Akt pathway. Cancer Res 68(23):9678–9685.  https://doi.org/10.1158/0008-5472.CAN-08-0969 CrossRefPubMedGoogle Scholar
  26. 26.
    Zhang XS, Hu YH, Gao HY, Lan XW, Xue YW (2017) Downregulation of Notch1 inhibits the invasion and metastasis of human gastric cancer cells SGC7901 and MKN74 in vitro through PTEN activation and dephosphorylation of Akt and FAK. Mol Med Rep 16(2):2318–2324.  https://doi.org/10.3892/mmr.2017.6791 CrossRefPubMedGoogle Scholar
  27. 27.
    Li L, Zhang J, Xiong N, Li S, Chen Y, Yang H, Wu C, Zeng H, Liu Y (2016) Notch-1 signaling activates NF-kappaB in human breast carcinoma MDA-MB-231 cells via PP2A-dependent Akt pathway. Med Oncol 33(4):33.  https://doi.org/10.1007/s12032-016-0747-7 CrossRefPubMedGoogle Scholar
  28. 28.
    Vlahos CJ, Matter WF, Hui KY, Brown RF (1994) A specific inhibitor of phosphatidylinositol 3-kinase, 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002). J Biol Chem 269(7):5241–5248PubMedGoogle Scholar
  29. 29.
    Zhu T, Hu X, Wei P, Shan G (2018) Molecular background of the regional lymph node metastasis of gastric cancer. Oncol Lett 15(3):3409–3414.  https://doi.org/10.3892/ol.2018.7813 CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Shimizu D, Kanda M, Kodera Y (2018) Emerging evidence of the molecular landscape specific for hematogenous metastasis from gastric cancer. World J Gastrointest Oncol 10(6):124–136.  https://doi.org/10.4251/wjgo.v10.i6.124 CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Zhong J, Chen Y, Wang LJ (2016) Emerging molecular basis of hematogenous metastasis in gastric cancer. World J Gastroenterol 22(8):2434–2440.  https://doi.org/10.3748/wjg.v22.i8.2434 CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Myong NH (2012) Loss of E-cadherin and acquisition of vimentin in epithelial–mesenchymal transition are noble indicators of uterine cervix cancer progression. Korean J Pathol 46(4):341–348.  https://doi.org/10.4132/KoreanJPathol.2012 CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Zheng ZH, Sun XJ, Zhou HT, Shang C, Ji H, Sun KL (2005) Analysis of metastasis suppressing function of E-cadherin in gastric cancer cells by RNAi. World J Gastroenterol 11(13):2000–2003.  https://doi.org/10.3748/wjg.v11.i13.2000 CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Tanaka M, Kitajima Y, Edakuni G, Sato S, Miyazaki K (2002) Abnormal expression of E-cadherin and beta-catenin may be a molecular marker of submucosal invasion and lymph node metastasis in early gastric cancer. Br J Surg 89(2):236–244.  https://doi.org/10.1046/j.0007-1323.2001.01985.x CrossRefPubMedGoogle Scholar
  35. 35.
    Liu CY, Lin HH, Tang MJ, Wang YK (2015) Vimentin contributes to epithelial–mesenchymal transition cancer cell mechanics by mediating cytoskeletal organization and focal adhesion maturation. Oncotarget 6(18):15966–15983.  https://doi.org/10.18632/oncotarget.3862 CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Shi Y, Shi H, Zhang B, Yan Y, Han X, Jiang W, Qian H, Xu W (2018) miR-373 suppresses gastric cancer metastasis by downregulating vimentin. Mol Med Rep 17(3):4027–4034.  https://doi.org/10.3892/mmr.2017.8291 CrossRefPubMedGoogle Scholar
  37. 37.
    Zhao K, He J, Wang YF, Jin SD, Fan Y, Fang N, Qian J, Xu TP, Guo RH (2019) EZH2-mediated epigenetic suppression of EphB3 inhibits gastric cancer proliferation and metastasis by affecting E-cadherin and vimentin expression. Gene 686:118–124.  https://doi.org/10.1016/j.gene.2018.11.015 CrossRefPubMedGoogle Scholar
  38. 38.
    Ren G, Tian Q, An Y, Feng B, Lu Y, Liang J, Li K, Shang Y, Nie Y, Wang X, Fan D (2012) Coronin 3 promotes gastric cancer metastasis via the up-regulation of MMP-9 and cathepsin K. Mol Cancer 11:67.  https://doi.org/10.1186/1476-4598-11-67 CrossRefPubMedPubMedCentralGoogle Scholar
  39. 39.
    Yoo YA, Kang MH, Lee HJ, Kim BH, Park JK, Kim HK, Kim JS, Oh SC (2011) Sonic hedgehog pathway promotes metastasis and lymphangiogenesis via activation of Akt, EMT, and MMP-9 pathway in gastric cancer. Cancer Res 71(22):7061–7070.  https://doi.org/10.1158/0008-5472.CAN-11-1338 CrossRefPubMedGoogle Scholar
  40. 40.
    Whelan JT, Kellogg A, Shewchuk BM, Hewan-Lowe K, Bertrand FE (2009) Notch-1 signaling is lost in prostate adenocarcinoma and promotes PTEN gene expression. J Cell Biochem 107(5):992–1001.  https://doi.org/10.1002/jcb.22199 CrossRefPubMedGoogle Scholar
  41. 41.
    Li X, Cao Y, Li M, Jin F (2018) Upregulation of HES1 promotes cell proliferation and invasion in breast cancer as a prognosis marker and therapy target via the Akt pathway and EMT process. J Cancer 9(4):757–766.  https://doi.org/10.7150/jca.22319 CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Wang SC, Lin XL, Wang HY, Qin YJ, Chen L, Li J, Jia JS, Shen HF, Yang S, Xie RY, Wei F, Gao F, Rong XX, Yang J, Zhao WT, Zhang TT, Shi JW, Yao KT, Luo WR, Sun Y, Xiao D (2015) Hes1 triggers epithelial–mesenchymal transition (EMT)-like cellular marker alterations and promotes invasion and metastasis of nasopharyngeal carcinoma by activating the PTEN/Akt pathway. Oncotarget 6(34):36713–36730.  https://doi.org/10.18632/oncotarget.5457 CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Sokolowski KM, Balamurugan M, Kunnimalaiyaan S, Wilson J, Gamblin TC, Kunnimalaiyaan M (2016) Role of Akt inhibition on Notch1 expression in hepatocellular carcinoma: potential role for dual targeted therapy. Am J Surg 211(4):755–760.  https://doi.org/10.1016/j.amjsurg.2015.11.029 CrossRefPubMedGoogle Scholar
  44. 44.
    Bedogni B, Warneke JA, Nickoloff BJ, Giaccia AJ, Powell MB (2008) Notch1 is an effector of Akt and hypoxia in melanoma development. J Clin Invest 118(11):3660–3670.  https://doi.org/10.1172/JCI36157 CrossRefPubMedPubMedCentralGoogle Scholar
  45. 45.
    Platonova N, Manzo T, Mirandola L, Colombo M, Calzavara E, Vigolo E, Cermisoni GC, De Simone D, Garavelli S, Cecchinato V, Lazzari E, Neri A, Chiaramonte R (2015) PI3K/Akt signaling inhibits NOTCH1 lysosome-mediated degradation. Genes Chromosomes Cancer 54(8):516–526.  https://doi.org/10.1002/gcc.22264 CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.College of Basic Medical SciencesHubei University of MedicineShiyanChina
  2. 2.Department of Rehabilitation Medicine, Taihe HospitalHubei University of MedicineShiyanChina
  3. 3.Department of Pathology, Shanghai Xuhui Central Hospital, Zhongshan-Xuhui Hospital, Shanghai Clinical CenterFudan University, CASShanghaiChina
  4. 4.Department of Neurosurgery, Taihe HospitalHubei University of MedicineShiyanChina

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