Top

Published in:

01-07-2016 | Original Article

Upregulation of CD44v6 contributes to acquired chemoresistance via the modulation of autophagy in colon cancer SW480 cells

Authors: Lin Lv, Hai-Guang Liu, Si-Yang Dong, Fan Yang, Qing-Xuan Wang, Gui-Long Guo, Yi-Fei Pan, Xiao-Hua Zhang

Published in: Tumor Biology | Issue 7/2016

Abstract

The CD44 isoform containing variant exon v6 (CD44v6) plays an important role in the progression, metastasis, and prognosis of colorectal cancer (CRC). Recently, it was found that CD44v6 is involved in acquired drug resistance. This study aimed to investigate the molecular mechanism of CD44v6 in the resistance of CRC cells to chemotherapy. A stable CD44v6 overexpression model in SW480 cells was established via lentiviral transduction. The chemosensitivity of cells to 5-fluorouracil (5-FU) and oxaliplatin (L-OHP) was determined by cell counting kit (CCK)-8, lactate dehydrogenase (LDH) release, and colony formation assays. Immunohistochemical staining of CD44v6 was performed in human CRC tissues. The key components in cell apoptosis, drug efflux and metabolism, mismatch repair, autophagy, epithelial–mesenchymal transition (EMT), and the PI3K–Akt and MAPK–Ras–Erk1/2 pathways were assessed using flow cytometry, quantitative real-time polymerase chain reaction (PCR), and western blot assays. The CD44v6 overexpression cells showed a higher viability, a lower LDH release rate, and an increased clonogenicity than the control cells under drug treatment. Moreover, overexpression of CD44v6 resulted in enhanced autophagy flux, EMT, and phosphorylation of Akt and Erk in the presence of drugs. Furthermore, high CD44v6 expression in the primary tumor was closely associated with an early recurrence in CRC patients who underwent curative surgery and adjuvant chemotherapy. In conclusion, overexpression of CD44v6 contributes to chemoresistance in SW480 cells under cytotoxic stress via the modulation of autophagy, EMT, and activation of the PI3K–Akt and MAPK–Ras–Erk pathways.

Cunningham D, Atkin W, Lenz HJ, Lynch HT, Minsky B, Nordlinger B, et al. Colorectal cancer. Lancet. 2010;375(9719):1030–47.CrossRefPubMed

Holohan C, Van Schaeybroeck S, Longley DB, Johnston PG. Cancer drug resistance: an evolving paradigm. Nat Rev Cancer. 2013;13(10):714–26. doi:10.1038/nrc3599.CrossRefPubMed

Longley DB, Johnston PG. Molecular mechanisms of drug resistance. J Pathol. 2005;205(2):275–92. doi:10.1002/path.1706.CrossRefPubMed

De Mattia E, Cecchin E, Toffoli G. Pharmacogenomics of intrinsic and acquired pharmacoresistance in colorectal cancer: toward targeted personalized therapy. Drug Resist Updat. 2015;20:39–70. doi:10.1016/j.drup.2015.05.003.CrossRefPubMed

Perez-Tomas R. Multidrug resistance: retrospect and prospects in anti-cancer drug treatment. Curr Med Chem. 2006;13(16):1859–76. doi:10.2174/092986706777585077.CrossRefPubMed

Glavinas H, Krajcsi P, Cserepes J, Sarkadi B. The role of ABC transporters in drug resistance, metabolism and toxicity. Curr Drug Deliv. 2004;1(1):27–42. doi:10.2174/1567201043480036.CrossRefPubMed

Townsend DM, Tew KD. The role of glutathione-S-transferase in anti-cancer drug resistance. Oncogene. 2003;22(47):7369–75. doi:10.1038/sj.onc.1206940.CrossRefPubMed

Giles GI, Sharma RP. Topoisomerase enzymes as therapeutic targets for cancer chemotherapy. Med Chem. 2005;1(4):383–94.CrossRefPubMed

Kirschner K, Melton DW. Multiple roles of the ERCC1-XPF endonuclease in DNA repair and resistance to anticancer drugs. Anticancer Res. 2010;30(9):3223–32.PubMed

10.

Lage H, Dietel M. Involvement of the DNA mismatch repair system in antineoplastic drug resistance. J Cancer Res Clin Oncol. 1999;125(3–4):156–65.CrossRefPubMed

11.

Rodriguez-Nieto S, Zhivotovsky B. Role of alterations in the apoptotic machinery in sensitivity of cancer cells to treatment. Curr Pharm Des. 2006;12(34):4411–25.CrossRefPubMed

12.

Lai K, Killingsworth MC, Lee CS. The significance of autophagy in colorectal cancer pathogenesis and implications for therapy. J Clin Pathol. 2014;67(10):854–8. doi:10.1136/jclinpath-2014-202529.CrossRefPubMed

13.

McCubrey JA, Steelman LS, Kempf CR, Chappell WH, Abrams SL, Stivala F, et al. Therapeutic resistance resulting from mutations in Raf/MEK/ERK and PI3K/PTEN/Akt/mTOR signaling pathways. J Cell Physiol. 2011;226(11):2762–81. doi:10.1002/jcp.22647.CrossRefPubMed

14.

Sui H, Zhu L, Deng WL, Li Q. Epithelial-mesenchymal transition and drug resistance: role, molecular mechanisms, and therapeutic strategies. Oncol Res Treat. 2014;37(10):584–9. doi:10.1159/000367802.CrossRefPubMed

15.

Ponta H, Sherman L, Herrlich PA. CD44: from adhesion molecules to signalling regulators. Nat Rev Mol Cell Biol. 2003;4(1):33–45. doi:10.1038/nrm1004.CrossRefPubMed

16.

Coppola D, Hyacinthe M, Fu L, Cantor AB, Karl R, Marcet J, et al. CD44V6 expression in human colorectal carcinoma. Hum Pathol. 1998;29(6):627–35.CrossRefPubMed

17.

Zlobec I, Gunthert U, Tornillo L, Iezzi G, Baumhoer D, Terracciano L, et al. Systematic assessment of the prognostic impact of membranous CD44v6 protein expression in colorectal cancer. Histopathology. 2009;55(5):564–75. doi:10.1111/j.1365-2559.2009.03421.x.CrossRefPubMed

18.

Todaro M, Gaggianesi M, Catalano V, Benfante A, Iovino F, Biffoni M, et al. CD44v6 is a marker of constitutive and reprogrammed cancer stem cells driving colon cancer metastasis. Cell Stem Cell. 2014;14(3):342–56. doi:10.1016/j.stem.2014.01.009.CrossRefPubMed

19.

Reinhold WC, Sunshine M, Liu H, Varma S, Kohn KW, Morris J, et al. Cell miner: a web-based suite of genomic and pharmacologic tools to explore transcript and drug patterns in the NCI-60 cell line set. Cancer Res. 2012;72(14):3499–511. doi:10.1158/0008-5472.CAN-12-1370.CrossRefPubMedPubMedCentral

20.

Larionov A, Krause A, Miller W. A standard curve based method for relative real time PCR data processing. BMC Bioinformatics. 2005;6:62. doi:10.1186/1471-2105-6-62.CrossRefPubMedPubMedCentral

21.

Klionsky DJ, Abdalla FC, Abeliovich H, Abraham RT, Acevedo-Arozena A, Adeli K, et al. Guidelines for the use and interpretation of assays for monitoring autophagy. Autophagy. 2012;8(4):445–544.CrossRefPubMedPubMedCentral

22.

Yanamoto S, Yamada S, Takahashi H, Naruse T, Matsushita Y, Ikeda H, et al. Expression of the cancer stem cell markers CD44v6 and ABCG2 in tongue cancer: effect of neoadjuvant chemotherapy on local recurrence. Int J Oncol. 2014;44(4):1153–62. doi:10.3892/ijo.2014.2289.PubMed

23.

Costa S, Terzano P, Bovicelli A, Martoni A, Angelelli B, Santini D, et al. CD44 isoform 6 (CD44v6) is a prognostic indicator of the response to neoadjuvant chemotherapy in cervical carcinoma. Gynecol Oncol. 2001;80(1):67–73. doi:10.1006/gyno.2000.6016.CrossRefPubMed

24.

Bendardaf R, Lamlum H, Ristamaki R, Pyrhonen S. CD44 variant 6 expression predicts response to treatment in advanced colorectal cancer. Oncol Rep. 2004;11(1):41–5.PubMed

25.

Niu RF, Zhang J, Huang JY. Expression of CD44v6 before and after chemotherapy in patients with breast cancer and its significance. Ai Zheng. 2002;21(1):71–4.PubMed

26.

Recio JA, Merlino G. Hepatocyte growth factor/scatter factor induces feedback up-regulation of CD44v6 in melanoma cells through Egr-1. Cancer Res. 2003;63(7):1576–82.PubMed

27.

Gao C, Guo H, Downey L, Marroquin C, Wei J, Kuo PC. Osteopontin-dependent CD44v6 expression and cell adhesion in HepG2 cells. Carcinogenesis. 2003;24(12):1871–8. doi:10.1093/carcin/bgg139.CrossRefPubMed

28.

Li J, Zha XM, Wang R, Li XD, Xu B, Xu YJ, et al. Regulation of CD44 expression by tumor necrosis factor-alpha and its potential role in breast cancer cell migration. Biomed Pharmacother. 2012;66(2):144–50. doi:10.1016/j.biopha.2011.11.021.CrossRefPubMed

29.

Quinones A, Dobberstein KU, Rainov NG. The egr-1 gene is induced by DNA-damaging agents and non-genotoxic drugs in both normal and neoplastic human cells. Life Sci. 2003;72(26):2975–92.CrossRefPubMed

30.

Damm S, Koefinger P, Stefan M, Wels C, Mehes G, Richtig E, et al. HGF-promoted motility in primary human melanocytes depends on CD44v6 regulated via NF-kappa B, Egr-1, and C/EBP-beta. J Invest Dermatol. 2010;130(7):1893–903. doi:10.1038/jid.2010.45.CrossRefPubMedPubMedCentral

31.

Hebbard L, Steffen A, Zawadzki V, Fieber C, Howells N, Moll J, et al. CD44 expression and regulation during mammary gland development and function. J Cell Sci. 2000;113(Pt 14):2619–30.PubMed

32.

Ni J, Cozzi PJ, Hao JL, Beretov J, Chang L, Duan W, et al. CD44 variant 6 is associated with prostate cancer metastasis and chemo-/radioresistance. Prostate. 2014;74(6):602–17. doi:10.1002/pros.22775.CrossRefPubMed

33.

Miletti-Gonzalez KE, Chen S, Muthukumaran N, Saglimbeni GN, Wu X, Yang J, et al. The CD44 receptor interacts with P-glycoprotein to promote cell migration and invasion in cancer. Cancer Res. 2005;65(15):6660–7. doi:10.1158/0008-5472.CAN-04-3478.CrossRefPubMed

34.

Liu CM, Chang CH, Yu CH, Hsu CC, Huang LL. Hyaluronan substratum induces multidrug resistance in human mesenchymal stem cells via CD44 signaling. Cell Tissue Res. 2009;336(3):465–75. doi:10.1007/s00441-009-0780-3.CrossRefPubMed

35.

Misra S, Ghatak S, Toole BP. Regulation of MDR1 expression and drug resistance by a positive feedback loop involving hyaluronan, phosphoinositide 3-kinase, and ErbB2. J Biol Chem. 2005;280(21):20310–5. doi:10.1074/jbc.M500737200.CrossRefPubMed

36.

Xu ZY, Tang JN, Xie HX, Du YA, Huang L, Yu PF, et al. 5-Fluorouracil chemotherapy of gastric cancer generates residual cells with properties of cancer stem cells. Int J Biol Sci. 2015;11(3):284–94. doi:10.7150/ijbs.10248.CrossRefPubMedPubMedCentral

37.

Mathew R, Karantza-Wadsworth V, White E. Role of autophagy in cancer. Nat Rev Cancer. 2007;7(12):961–7. doi:10.1038/nrc2254.CrossRefPubMedPubMedCentral

38.

Li J, Hou N, Faried A, Tsutsumi S, Takeuchi T, Kuwano H. Inhibition of autophagy by 3-MA enhances the effect of 5-FU-induced apoptosis in colon cancer cells. Ann Surg Oncol. 2009;16(3):761–71. doi:10.1245/s10434-008-0260-0.CrossRefPubMed

39.

Sasaki K, Tsuno NH, Sunami E, Kawai K, Hongo K, Hiyoshi M, et al. Resistance of colon cancer to 5-fluorouracil may be overcome by combination with chloroquine, an in vivo study. Anticancer Drugs. 2012;23(7):675–82. doi:10.1097/CAD.0b013e328353f8c7.CrossRefPubMed

40.

Sasaki K, Tsuno NH, Sunami E, Tsurita G, Kawai K, Okaji Y, et al. Chloroquine potentiates the anti-cancer effect of 5-fluorouracil on colon cancer cells. BMC Cancer. 2010;10:370. doi:10.1186/1471-2407-10-370.CrossRefPubMedPubMedCentral

41.

Park JM, Huang S, Wu TT, Foster NR, Sinicrope FA. Prognostic impact of Beclin 1, p62/sequestosome 1 and LC3 protein expression in colon carcinomas from patients receiving 5-fluorouracil as adjuvant chemotherapy. Cancer Biol Ther. 2013;14(2):100–7. doi:10.4161/cbt.22954.CrossRefPubMedPubMedCentral

42.

Zaanan A, Park JM, Tougeron D, Huang S, Wu TT, Foster NR, et al. Association of beclin 1 expression with response to neoadjuvant chemoradiation therapy in patients with locally advanced rectal carcinoma. Int J Cancer. 2015;137(6):1498–502. doi:10.1002/ijc.29496.CrossRefPubMedPubMedCentral

43.

Ogier-Denis E, Pattingre S, El Benna J, Codogno P. Erk1/2-dependent phosphorylation of Galpha-interacting protein stimulates its GTPase accelerating activity and autophagy in human colon cancer cells. J Biol Chem. 2000;275(50):39090–5. doi:10.1074/jbc.M006198200.CrossRefPubMed

44.

Levine B, Sinha S, Kroemer G. Bcl-2 family members—dual regulators of apoptosis and autophagy. Autophagy. 2008;4(5):600–6.CrossRefPubMedCentral

45.

Graziani A, Gramaglia D, Cantley LC, Comoglio PM. The tyrosine-phosphorylated hepatocyte growth factor/scatter factor receptor associates with phosphatidylinositol 3-kinase. J Biol Chem. 1991;266(33):22087–90.PubMed

46.

Orian-Rousseau V, Chen L, Sleeman JP, Herrlich P, Ponta H. CD44 is required for two consecutive steps in HGF/c-Met signaling. Genes Dev. 2002;16(23):3074–86. doi:10.1101/gad.242602.CrossRefPubMedPubMedCentral

47.

Klingbeil P, Marhaba R, Jung T, Kirmse R, Ludwig T, Zoller M. CD44 variant isoforms promote metastasis formation by a tumor cell-matrix cross-talk that supports adhesion and apoptosis resistance. Mol Cancer Res. 2009;7(2):168–79. doi:10.1158/1541-7786.MCR-08-0207.CrossRefPubMed

48.

Jung T, Gross W, Zoller M. CD44v6 coordinates tumor matrix-triggered motility and apoptosis resistance. J Biol Chem. 2011;286(18):15862–74. doi:10.1074/jbc.M110.208421.CrossRefPubMedPubMedCentral

Title: Upregulation of CD44v6 contributes to acquired chemoresistance via the modulation of autophagy in colon cancer SW480 cells
Authors: Lin Lv
Hai-Guang Liu
Si-Yang Dong
Fan Yang
Qing-Xuan Wang
Gui-Long Guo
Yi-Fei Pan
Xiao-Hua Zhang
Publication date: 01-07-2016
Publisher: Springer Netherlands
Published in: Tumor Biology / Issue 7/2016
Print ISSN: 1010-4283
Electronic ISSN: 1423-0380
DOI: https://doi.org/10.1007/s13277-015-4755-6

Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras

Prof. Fabrice Barlesi

Developed by: Springer Medicine

At a glance: The STEP trials

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 7/2016

B7-H3 increases thymidylate synthase expression via the PI3k-Akt pathway

Monitoring the treatment outcome in endometrial cancer patients by CEA and TATI

Elevated growth differentiation factor 15 expression predicts poor prognosis in epithelial ovarian cancer patients

Evaluation of RIP1K and RIP3K expressions in the malignant and benign breast tumors

Total tumor volume predicts survival following liver resection in patients with hepatocellular carcinoma

Comparison of efficacy and safety of three different chemotherapy regimens delivered with concomitant radiotherapy in inoperable stage III non-small cell lung cancer patients

Keynote webinar | Spotlight on antibody–drug conjugates in cancer