Advertisement

Journal of Cancer Research and Clinical Oncology

, Volume 145, Issue 11, pp 2649–2661 | Cite as

CD44 splice variant (CD44v3) promotes progression of urothelial carcinoma of bladder through Akt/ERK/STAT3 pathways: novel therapeutic approach

  • Vivek Anand
  • Madhuram Khandelwal
  • Sandeep Appunni
  • Nidhi Gupta
  • Amlesh Seth
  • Prabhjot Singh
  • Sandeep Mathur
  • Alpana SharmaEmail author
Original Article – Cancer Research

Abstract

Purpose

The incidence of Urothelial carcinoma of bladder (UBC) is gradually increasing by changing lifestyle and environment. The development of a tumor has been noted to be accompanied by modifications in the extracellular matrix (ECM) consisting of CD44, hyaluronic acid (HA) and its family members. The importance of CD44 splice variants and HA family members has been studied in UBC.

Methods

The cohort of study included 50 UBC patients undergoing radical cystectomy and 50 healthy subjects. The molecular expression of CD44 and HA family members was determined. Effect of CD44 variant-specific silencing on downstream signaling in HT1376 cells was investigated. Combinatorial treatment of 4-MU (4-methylumbelliferone) with cisplatin or doxorubicin on chemosensitivity was also explored.

Results

Higher expression of HA, HAS2, and CD44 was observed in Indian UBC patients which also showed the trend with severity of disease. Splice variant assessment of CD44 demonstrated the distinct role of CD44v3 and CD44v6 in bladder cancer progression. shRNA-mediated downregulation of CD44v3 showed an increase effect on cell cycle, apoptosis and multiple downstream signaling cascade including pAkt, pERK and pSTAT3. Furthermore, 4-MU, an HA synthesis inhibitor, observed to complement the effect of Cisplatin or Doxorubicin by enhancing the chemosensitivity of bladder cancer cells.

Conclusions

Our findings exhibit involvement of CD44 splice variants and HA family members in UBC and significance of 4-MU in enhancing chemosensitivity suggesting their novel therapeutic importance in disease therapeutics.

Keywords

Bladder cancer Hyaluronic acid Hyaluronic acid synthase CD44 CD44 variant 3 4-Methylumbelliferone 

Notes

Acknowledgements

We acknowledge Indian Council for Medical Research (ICMR) for the grant to carry out research work. ICMR Grant No.: 5/13/50/10/NCD-III. All the authors have read the journal’s authorship agreement and policy.

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.

Supplementary material

432_2019_3024_MOESM1_ESM.doc (556 kb)
Supplementary material 1 (DOC 556 kb)

References

  1. Adamia S, Pilarski PM, Belch AR, Pilarski LM (2013) Aberrant splicing, hyaluronan synthases and intracellular hyaluronan as drivers of oncogenesis and potential drug targets. Curr Cancer Drug Targets 13(4):347–361PubMedCrossRefGoogle Scholar
  2. Akisik E, Bavbek S, Dalay N (2002) CD44 variant exons in leukemia and lymphoma. Pathol Oncol Res 8(1):36–40PubMedCrossRefGoogle Scholar
  3. Amirghofran Z, Jalali SA, Hosseini SV, Vasei M, Sabayan B, Ghaderi A (2008) Evaluation of CD44 and CD44v6 in colorectal carcinoma patients: soluble forms in relation to tumor tissue expression and metastasis. J Gastrointest Cancer 39(1–4):73–78PubMedCrossRefGoogle Scholar
  4. Bánky B, Rásó-Barnett L, Barbai T, Tímár J, Becságh P, Rásó E (2012) Characteristics of CD44 alternative splice pattern in the course of human colorectal adenocarcinoma progression. Mol. Cancer 11:83PubMedPubMedCentralCrossRefGoogle Scholar
  5. Basakran NS (2015) CD44 as a potential diagnostic tumor marker. Saudi Med J 36(3):273–279PubMedPubMedCentralCrossRefGoogle Scholar
  6. Bernert B, Porsch H, Heldin P (2011) Hyaluronan synthase 2 (HAS2) promotes breast cancer cell invasion by suppression of tissue metalloproteinase inhibitor 1 (TIMP-1). J Biol Chem 286(49):42349–42359PubMedPubMedCentralCrossRefGoogle Scholar
  7. Chen C, Zhao S, Karnad A, Freeman JW (2018) The biology and role of CD44 in cancer progression: therapeutic implications. J Hematol Oncol 11(1):64PubMedPubMedCentralCrossRefGoogle Scholar
  8. Cheng XB, Sato N, Kohi S, Yamaguchi K (2013) Prognostic impact of hyaluronan and its regulators in pancreatic ductal adenocarcinoma. PLoS One 8(11):e80765PubMedPubMedCentralCrossRefGoogle Scholar
  9. Gee K, Kryworuchko M, Kumar A (2004) Recent advances in the regulation of CD44 expression and its role in inflammation and autoimmune diseases. Arch Immunol Ther Exp (Warsz) 52(1):13–26Google Scholar
  10. Ghatak S, Misra S, Toole BP (2005) Hyaluronan constitutively regulates ErbB2 phosphorylation and signaling complex formation in carcinoma cells. J Biol Chem 280(10):8875–8883PubMedCrossRefGoogle Scholar
  11. Hamilton SR, Fard SF, Paiwand FF, Tolg C, Veiseh M, Wang C, McCarthy JB, Bissell MJ, Koropatnick J, Turley EA (2007) The hyaluronan receptors CD44 and Rhamm (CD168) form complexes with ERK1,2 that sustain high basal motility in breast cancer cells. J Biol Chem 282(22):16667–16680PubMedPubMedCentralCrossRefGoogle Scholar
  12. He Q, Lesley J, Hyman R, Ishihara K, Kincade PW (1992) Molecular isoforms of murine CD44 and evidence that the membrane proximal domain is not critical for hyaluronate recognition. J Cell Biol 119(6):1711–1719PubMedCrossRefGoogle Scholar
  13. Herishanu Y, Gibellini F, Njuguna N, Hazan-Halevy I, Farooqui M, Bern S, Keyvanfar K, Lee E, Wilson W, Wiestner A (2011) Activation of CD44, a receptor for extracellular matrix components, protects chronic lymphocytic leukemia cells from spontaneous and drug induced apoptosis through MCL-1. Leuk Lymphoma 52(9):1758–1769PubMedPubMedCentralCrossRefGoogle Scholar
  14. Ilangumaran S, Briol A, Hoessli DC (1998) CD44 selectively associates with active Src family protein tyrosine kinases Lck and Fyn in glycosphingolipid-rich plasma membrane domains of human peripheral blood lymphocytes. Blood 91(10):3901–3908PubMedCrossRefGoogle Scholar
  15. Jamshidian H, Hashemi M, Nowroozi MR, Ayati M, Bonyadi M, Najjaran Tousi V (2014) Sensitivity and specificity of urinary hyaluronic acid and hyaluronidase in detection of bladder transitional cell carcinoma. Urol J 11(1):1232–1237PubMedGoogle Scholar
  16. Jemal A, Murray T, Ward E, Samuels A, Tiwari RC, Ghafoor A, Feuer EJ, Thun MJ (2005) Cancer statistics, 2005. CA Cancer J Clin 55(1):10–30PubMedCrossRefGoogle Scholar
  17. Knudsen LM, Rasmussen T, Jensen L, Johnsen HE (1999) Reduced bone marrow stem cell pool and progenitor mobilisation in multiple myeloma after melphalan treatment. Med Oncol 16(4):245–254PubMedCrossRefGoogle Scholar
  18. Kobayashi K, Matsumoto H, Matsuyama H, Fujii N, Inoue R, Yamamoto Y, Nagao K (2016) Clinical significance of CD44 variant 9 expression as a prognostic indicator in bladder cancer. Oncol Rep 36(5):2852–2860PubMedCrossRefGoogle Scholar
  19. Kosaki R, Watanabe K, Yamaguchi Y (1999) Overproduction of hyaluronan by expression of the hyaluronan synthase Has2 enhances anchorage-independent growth and tumorigenicity. Cancer Res 59(5):1141–1145PubMedGoogle Scholar
  20. Kouvidi K, Berdiaki A, Nikitovic D, Katonis P, Afratis N, Hascall VC, Karamanos NK, Tzanakakis GN (2011) Role of receptor for hyaluronic acid-mediated motility (RHAMM) in low molecular weight hyaluronan (LMWHA)-mediated fibrosarcoma cell adhesion. J Biol Chem 286(44):38509–38520PubMedPubMedCentralCrossRefGoogle Scholar
  21. Kramer MW, Escudero DO, Lokeshwar SD, Golshani R, Ekwenna OO, Acosta K, Merseburger AS, Soloway M, Lokeshwar VB (2011) Association of hyaluronic acid family members (HAS1, HAS2, and HYAL-1) with bladder cancer diagnosis and prognosis. Cancer 117(6):1197–1209PubMedCrossRefGoogle Scholar
  22. Lamouille S, Xu J, Derynck R (2014) Molecular mechanisms of epithelial-mesenchymal transition. Nat Rev Mol Cell Biol 15(3):178–196PubMedPubMedCentralCrossRefGoogle Scholar
  23. Li P, Xiang T, Li H, Li Q, Yang B, Huang J, Zhang X, Shi Y, Tan J, Ren G (2015) Hyaluronan synthase 2 overexpression is correlated with the tumorigenesis and metastasis of human breast cancer. Int J Clin Exp Pathol 8(10):12101–12114PubMedPubMedCentralGoogle Scholar
  24. Lokeshwar VB, Mirza S, Jordan A (2014) Targeting hyaluronic acid family for cancer chemoprevention and therapy. Adv Cancer Res 123:35–65PubMedPubMedCentralCrossRefGoogle Scholar
  25. Mayr L, Pirker C, Lötsch D, Van Schoonhoven S, Windhager R, Englinger B, Berger W, Kubista B (2017) CD44 drives aggressiveness and chemoresistance of a metastatic human osteosarcoma xenograft model. Oncotarget 8(69):114095–114108PubMedPubMedCentralCrossRefGoogle Scholar
  26. Meran S, Luo DD, Simpson R, Martin J, Wells A, Steadman R, Phillips AO (2011) Hyaluronan facilitates transforming growth factor-β1-dependent proliferation via CD44 and epidermal growth factor receptor interaction. J Biol Chem 286(20):17618–17630PubMedPubMedCentralCrossRefGoogle Scholar
  27. Misra S, Toole BP, Ghatak S (2006) Hyaluronan constitutively regulates activation of multiple receptor tyrosine kinases in epithelial and carcinoma cells. J Biol Chem 281(46):34936–34941PubMedCrossRefGoogle Scholar
  28. Morath I, Jung C, Lévêque R, Linfeng C, Toillon RA, Warth A, Orian-Rousseau V (2018) Differential recruitment of CD44 isoforms by ErbB ligands reveals an involvement of CD44 in breast cancer. Oncogene 37(11):1472–1484PubMedCrossRefGoogle Scholar
  29. Morera DS, Hennig MS, Talukder A, Lokeshwar SD, Wang J, Garcia-Roig M, Ortiz N, Yates TJ, Lopez LE, Kallifatidis G et al (2017) Hyaluronic acid family in bladder cancer: potential prognostic biomarkers and therapeutic targets. Br J Cancer 117(10):1507–1517PubMedPubMedCentralCrossRefGoogle Scholar
  30. Olsson E, Honeth G, Bendahl PO, Saal LH, Gruvberger-Saal S, Ringnér M, Vallon-Christersson J, Jönsson G, Holm K, Lövgren K et al (2011) CD44 isoforms are heterogeneously expressed in breast cancer and correlate with tumor subtypes and cancer stem cell markers. BMC Cancer 11:418PubMedPubMedCentralCrossRefGoogle Scholar
  31. Park SH, Lee Y, Han SH, Kwon SY, Kwon OS, Kim SS, Kim JH, Park YH, Lee JN, Bang SM et al (2006) Systemic chemotherapy with doxorubicin, cisplatin and capecitabine for metastatic hepatocellular carcinoma. BMC Cancer 6:3PubMedPubMedCentralCrossRefGoogle Scholar
  32. Parkin DM (2008) The global burden of urinary bladder cancer. Scand J Urol Nephrol Suppl 218:12–20CrossRefGoogle Scholar
  33. Ploeg M, Aben KKH, Kiemeney LA (2009) The present and future burden of urinary bladder cancer in the world. World J Urol 27(3):289–293PubMedPubMedCentralCrossRefGoogle Scholar
  34. Saito T, Tamura D, Nakamura T, Makita Y, Ariyama H, Komiyama K, Yoshihara T, Asano R (2013) 4-methylumbelliferone leads to growth arrest and apoptosis in canine mammary tumor cells. Oncol Rep 29(1):335–342PubMedCrossRefGoogle Scholar
  35. Sato N, Maehara N, Goggins M (2004) Gene expression profiling of tumor-stromal interactions between pancreatic cancer cells and stromal fibroblasts. Cancer Res 64(19):6950–6956PubMedCrossRefGoogle Scholar
  36. Stern R, Asari AA, Sugahara KN (2006) Hyaluronan fragments: an information-rich system. Eur J Cell Biol 85(8):699–715PubMedCrossRefGoogle Scholar
  37. Subramaniam V, Gardner H, Jothy S (2007) Soluble CD44 secretion contributes to the acquisition of aggressive tumor phenotype in human colon cancer cells. Exp Mol Pathol 83(3):341–346PubMedCrossRefGoogle Scholar
  38. Toole BP (2004) Hyaluronan: from extracellular glue to pericellular cue. Nat Rev Cancer 4(7):528–539PubMedCrossRefGoogle Scholar
  39. Toole BP, Hascall VC (2002) Hyaluronan and tumor growth. Am J Pathol 161(3):745–747PubMedPubMedCentralCrossRefGoogle Scholar
  40. Turley EA, Noble PW, Bourguignon LYW (2002) Signaling properties of hyaluronan receptors. J Biol Chem 277(7):4589–4592PubMedCrossRefGoogle Scholar
  41. Urakawa H, Nishida Y, Knudson W, Knudson CB, Arai E, Kozawa E, Futamura N, Wasa J, Ishiguro N (2012) Therapeutic potential of hyaluronan oligosaccharides for bone metastasis of breast cancer. J Orthop Res 30(4):662–672PubMedCrossRefGoogle Scholar
  42. Van Hal NL, Van Dongen GA, Ten Brink CB, Heider KH, Rech-Weichselbraun I, Snow GB, Brakenhoff RH (1999) Evaluation of soluble CD44v6 as a potential serum marker for head and neck squamous cell carcinoma. Clin Cancer Res 5(11):3534–3541PubMedGoogle Scholar
  43. Williams K, Motiani K, Giridhar PV, Kasper S (2013) CD44 integrates signaling in normal stem cell, cancer stem cell and (pre)metastatic niches. Exp Biol Med (Maywood) 238(3):324–338CrossRefGoogle Scholar

Copyright information

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

Authors and Affiliations

  1. 1.Department of BiochemistryAll India Institute of Medical Sciences (AIIMS)New DelhiIndia
  2. 2.Department of UrologyAll India Institute of Medical Sciences (AIIMS)New DelhiIndia
  3. 3.Department of PathologyAll India Institute of Medical Sciences (AIIMS)New DelhiIndia

Personalised recommendations