Clinical and Experimental Medicine

, Volume 15, Issue 2, pp 137–144 | Cite as

microRNA-218 increase the sensitivity of gastrointestinal stromal tumor to imatinib through PI3K/AKT pathway

  • Rong Fan
  • Jie Zhong
  • Sichang Zheng
  • Zhengting Wang
  • Ying Xu
  • Shuyi Li
  • Jie Zhou
  • Fei Yuan
Original Article


To detect the expressions of microRNA-218 (miR-218) in an imatinib mesylate-sensitive human gastrointestinal stromal tumor (GIST) cells (GIST882) and an imatinib mesylate-resistant cell line (GIST430) and explore the roles of miR-218 and GIST cells in the sensitivity of gastrointestinal stromal tumor to imatinib mesylate and its potential signaling pathways, with an attempt to provide new insights for the treatment of GIST. The GIST cell lines (GIST882 and GIST430) were cultured in vitro. Quantitative real-time PCR (qRT-PCR) was utilized to determine the expression profiles of miR-218 in both GIST cell lines. Forty-eight hours after the transfection of the miR-218 mimic or miR-218 inhibitor in the GIST cells, the changes in the expression of miR-218 in the GIST cells were detected with qRT-PCR. The effects of the ectopic expression of miR-218 in GIST882 or GIST430 cells on the imatinib mesylate-induced GIST cell viability were determined by MTT. The effects of miR-218 ectopic expression on the apoptosis of imatinib mesylate-induce GIST cells were determined by Annexin V/PI double staining method and flow cytometry. The effects of miR-218 ectopic expression on the AKT and phospho-AKT (p-AKT) expressions of imatinib mesylate-induce GIST cells were determined by Western blot and flow cytometry with the PI3K pathway inhibitor Wortmannin. As shown by qRT-PCR, compared with that in the imatinib mesylate-sensitive GIST882, the expression of miR-218 in imatinib mesylate-resistant GIST430 was significantly decreased (P < 0.01). Compared with the control group, the expression of miR-218 significantly increased in the GIST882 48 h after the transfection of miR-218 mimic (P < 0.01) and significantly declined after the transfection of miR-218 inhibitor (P < 0.01). As shown by MTT and flow cytometry, after the expression of miR-218 was inhibited in GIST882 under the effect of imatinib mesylate, the cell viability significantly increased (P < 0.01) and the number of apoptotic cells significantly decreased (P < 0.05); on the contrary, the over-expression of miR-218 in GIST430 under the effect of imatinib mesylate resulted in the significantly decreased cell viability (P < 0.01) and the significantly increased number of apoptotic cells (P < 0.05). Western blot and flow cytometry showed that, in comparison to the control group, Wortmannin could significantly inhibit the expression of p-AKT in GIST430 cells (P < 0.01) and stimulated apoptosis (P < 0.01). The expression of miR-218 is down-regulated in an imatinib mesylate-resistant GIST cell line (GIST430), whereas miR-218 over-expression can improve the sensitivity of GIST cells to imatinib mesylate, with PI3K/AKT signaling pathway possibly involved in the mechanism.


GIST miR-218 Imatinib mesylate Apoptosis PI3K/AKT 



This study was supported by Key Project of Science and Technology Commission of Shanghai Municipality (No. 13411950900).

Conflict of interest



  1. 1.
    Datar M, Khanna R (2012) Inpatient burden of gastrointestinal stromal tumors in the United States. J Gastrointest Oncol 3(4):335–341PubMedPubMedCentralGoogle Scholar
  2. 2.
    Hodges K, Kennedy L, Meng F, Alpini G, Francis H (2012) Mast cells, disease and gastrointestinal cancer: a comprehensive review of recent findings. Transl Gastrointest Cancer 1(2):138–150PubMedPubMedCentralGoogle Scholar
  3. 3.
    Halpern J, Kim YJ, Sultana R, Villani G (2012) Effectiveness of radiation therapy in GIST: a case report. J Gastrointest Oncol 3(2):143–146PubMedPubMedCentralGoogle Scholar
  4. 4.
    McDaniel K, Correa R, Zhou T, Johnson C, Francis H, Glaser S, Venter J, Alpini G, Meng F (2013) Functional role of microvesicles in gastrointestinal malignancies. Ann Transl Med 1(1):4PubMedPubMedCentralGoogle Scholar
  5. 5.
    Kanda T (2013) Criminal or bystander: imatinib and second primary malignancy in GIST patients. Chin J Cancer Res 25(5):490–492PubMedPubMedCentralGoogle Scholar
  6. 6.
    Kinross KM, Sheppard KE, Pearson RB, Phillips WA (2012) Targeting cancer with PI3K pathway inhibitors: who to aim at? Transl Cancer Res 1(2):119–121Google Scholar
  7. 7.
    Sun X, Wang J, Yang G (2012) Surgical treatment of esophageal leiomyoma larger than 5 cm in diameter: a case report and review of the literature. J Thorac Dis 4(3):323–326CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Zhao X, Yue C (2012) Gastrointestinal stromal tumor. J Gastrointest Oncol 3(3):189–208PubMedPubMedCentralGoogle Scholar
  9. 9.
    Eisenberg BL, Pipas JM (2012) Gastrointestinal stromal tumor—background, pathology, treatment. Hematol Oncol Clin N Am 26(6):1239–1259CrossRefGoogle Scholar
  10. 10.
    Kee D, Zalcberg JR (2012) Current and emerging strategies for the management of imatinib-refractory advanced gastrointestinal stromal tumors. Ther Adv Med Oncol 4(5):255–270CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Nannini M, Biasco G, Astolfi A et al (2013) An overview on molecular biology of KIT/PDGFRA wild type (WT) gastrointestinal stromal tumours (GIST). J Med Genet 50(10):653–661CrossRefPubMedGoogle Scholar
  12. 12.
    Wang C, Jin MS, Zou YB et al (2013) Diagnostic significance of DOG-1 and PKC-theta expression and c-Kit/PDGFRA mutations in gastrointestinal stromal tumours. Scand J Gastroenterol 48(9):1055–1065CrossRefPubMedGoogle Scholar
  13. 13.
    O’Brien KM, Orlow I, Antonescu CR et al (2013) Gastrointestinal stromal tumors, somatic mutations and candidate genetic risk variants. PLoS One 8(4):e62119CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Tan CJ, Yang JL, Crowe P, Goldstein D (2013) Targeted therapy in soft tissue sarcoma: a novel direction in therapeutics. Chin Clin Oncol 2(3):22PubMedGoogle Scholar
  15. 15.
    Rutkowski P, Bylina E, Klimczak A et al (2012) The outcome and predictive factors of sunitinib therapy in advanced gastrointestinal stromal tumors (GIST) after imatinib failure-one institution study. BMC Cancer 12:107CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Joensuu H (2013) Gastrointestinal stromal tumors: risk assessment and adjuvant therapy. Hematol Oncol Clin N Am 27(5):889–904CrossRefGoogle Scholar
  17. 17.
    Tirumani SH, Jagannathan JP, Krajewski KM et al (2013) Imatinib and beyond in gastrointestinal stromal tumors: a radiologist’s perspective. AJR Am J Roentgenol 201(4):801–810CrossRefPubMedGoogle Scholar
  18. 18.
    An HJ, Ryu MH, Ryoo BY et al (2013) The effects of surgical cytoreduction prior to imatinib therapy on the prognosis of patients with advanced GIST. Ann Surg Oncol 20(13):4212–4218Google Scholar
  19. 19.
    Fujimoto Y, Akiyoshi T, Konishi T et al (2013) Laparoscopic sphincter-preserving surgery (intersphincteric resection) after neoadjuvant imatinib treatment for gastrointestinal stromal tumor (GIST) of the rectum. Int J Colorectal Dis 29(1):111–116Google Scholar
  20. 20.
    Das D, Ganguly S, Deb AR et al (2013) Neoodjuvant imatinib mesylate for advanced primary and metastactic/recurrent gastro-intestinal stromal tumour (GIST). J Indian Med Assoc 111(1):21–23PubMedGoogle Scholar
  21. 21.
    Falor A, Arrington AK, Luu C et al (2013) Massive intra-abdominal imatinib-resistant gastrointestinal stromal tumor in a 21-year-old male. Case Rep Med 2013:373981PubMedPubMedCentralGoogle Scholar
  22. 22.
    Linch M, Claus J, Benson C (2013) Update on imatinib for gastrointestinal stromal tumors: duration of treatment. Onco Targets Ther 6:1011–1023PubMedPubMedCentralGoogle Scholar
  23. 23.
    Blay JY, Rutkowski P (2013) Adherence to imatinib therapy in patients with gastrointestinal stromal tumors. Cancer Treat Rev 40(2):242–247Google Scholar
  24. 24.
    Wang Q, Wei L, Guan X et al (2013) Briefing in family characteristics of microRNAs and their applications in cancer research. Biochim Biophys Acta 1844(1 Pt B):191–197Google Scholar
  25. 25.
    Kaplan BB, Kar AN, Gioio AE et al (2013) MicroRNAs in the axon and presynaptic nerve terminal. Front Cell Neurosci 7:126CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Li M, Fu W, Wo L et al (2013) miR-128 and its target genes in tumorigenesis and metastasis. Exp Cell Res 319(20):3059–3064Google Scholar
  27. 27.
    Leite-Moreira AM, Lourenco AP, Falcao-Pires I et al (2013) Pivotal role of microRNAs in cardiac physiology and heart failure. Drug Discov Today 18(23–24):1243–1249Google Scholar
  28. 28.
    Piva R, Spandidos DA, Gambari R (2013) From microRNA functions to microRNA therapeutics: novel targets and novel drugs in breast cancer research and treatment (Review). Int J Oncol 43(4):985–994PubMedPubMedCentralGoogle Scholar
  29. 29.
    Shi Y, Wang CZ, Hou YY et al (2013) Screening of differentially expressed microRNAs in borderline and malignant gastrointestinal stromal tumors. Zhonghua Bing Li Xue Za Zhi 42(1):20–25PubMedGoogle Scholar
  30. 30.
    Tu Y, Gao X, Li G et al (2013) MicroRNA-218 inhibits glioma invasion, migration, proliferation and cancer stem-like cell self-renewal by targeting the polycomb group gene Bmi1. Cancer Res 73(19):6046–6055Google Scholar
  31. 31.
    Jin J, Cai L, Liu ZM et al (2013) miRNA-218 inhibits osteosarcoma cell migration and invasion by down-regulating of TIAM1, MMP2 and MMP9. Asian Pac J Cancer Prev 14(6):3681–3684CrossRefPubMedGoogle Scholar
  32. 32.
    Li J, Ping Z, Ning H (2012) MiR-218 impairs tumor growth and increases chemo-sensitivity to cisplatin in cervical cancer. Int J Mol Sci 13(12):16053–16064CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Tuveson DA, Willis NA, Jacks T et al (2001) STI571 inactivation of the gastrointestinal stromal tumor c-KIT oncoprotein: biological and clinical implications. Oncogene 20(36):5054–5058CrossRefPubMedGoogle Scholar
  34. 34.
    Reichardt P, Joensuu H, Blay JY (2013) New fronts in the adjuvant treatment of GIST. Cancer Chemother Pharmacol 72(4):715–723CrossRefPubMedGoogle Scholar
  35. 35.
    Kwong LN, Davies MA (2013) Navigating the therapeutic complexity of PI3K pathway inhibition in melanoma. Clin Cancer Res 19(19):5310–5319CrossRefPubMedGoogle Scholar
  36. 36.
    Kitagishi Y, Matsuda S (2013) Diets involved in PPAR and PI3K/AKT/PTEN pathway may contribute to neuroprotection in a traumatic brain injury. Alzheimers Res Ther 5(5):42CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    Siegfried Z, Bonomi S, Ghigna C et al (2013) Regulation of the Ras-MAPK and PI3K-mTOR signalling pathways by alternative splicing in cancer. Int J Cell Biol 2013:568931CrossRefPubMedPubMedCentralGoogle Scholar
  38. 38.
    Blume-Jensen P, Jiang G, Hyman R et al (2000) Kit/stem cell factor receptor-induced activation of phosphatidylinositol 3′-kinase is essential for male fertility. Nat Genet 24(2):157–162CrossRefPubMedGoogle Scholar
  39. 39.
    Gibbons SJ, Rich A, Distad MA et al (2003) Kit/stem cell factor receptor-induced phosphatidylinositol 3′-kinase signalling is not required for normal development and function of interstitial cells of Cajal in mouse gastrointestinal tract. Neurogastroenterol Motil 15(6):643–653CrossRefPubMedGoogle Scholar
  40. 40.
    Rajendra R, Pollack SM, Jones RL (2013) Management of gastrointestinal stromal tumors. Future Oncol 9(2):193–206CrossRefPubMedGoogle Scholar
  41. 41.
    Floris G, Wozniak A, Sciot R et al (2013) A potent combination of the novel PI3K inhibitor, GDC-0941, with imatinib in gastrointestinal stromal tumor xenografts: long-lasting responses after treatment withdrawal. Clin Cancer Res 19(3):620–630CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag Italia 2014

Authors and Affiliations

  • Rong Fan
    • 1
  • Jie Zhong
    • 1
  • Sichang Zheng
    • 1
  • Zhengting Wang
    • 1
  • Ying Xu
    • 1
  • Shuyi Li
    • 1
  • Jie Zhou
    • 1
  • Fei Yuan
    • 2
  1. 1.Department of Gastroenterology, Shanghai Ruijin HospitalShanghai Jiao Tong University School of MedicineShanghaiChina
  2. 2.Department of Pathology, Shanghai Ruijin HospitalShanghai Jiao Tong University School of MedicineShanghaiChina

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