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

  • Original Article – Cancer Research
  • Published:
Journal of Cancer Research and Clinical Oncology Aims and scope Submit manuscript

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.

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Fig. 1

taken as median of the total study samples. Protein expressions (immunoblotting) of HAS1, HAS2 and CD44 in tissues (b) and their densitometry analysis (c) done using image-J software

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References

  • 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):347–361

    Article  CAS  PubMed  Google Scholar 

  • Akisik E, Bavbek S, Dalay N (2002) CD44 variant exons in leukemia and lymphoma. Pathol Oncol Res 8(1):36–40

    Article  CAS  PubMed  Google Scholar 

  • 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):73–78

    Article  CAS  PubMed  Google Scholar 

  • 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 11:83

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Basakran NS (2015) CD44 as a potential diagnostic tumor marker. Saudi Med J 36(3):273–279

    Article  PubMed  PubMed Central  Google Scholar 

  • 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):42349–42359

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • 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):64

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Cheng XB, Sato N, Kohi S, Yamaguchi K (2013) Prognostic impact of hyaluronan and its regulators in pancreatic ductal adenocarcinoma. PLoS One 8(11):e80765

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • 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):13–26

    CAS  Google Scholar 

  • Ghatak S, Misra S, Toole BP (2005) Hyaluronan constitutively regulates ErbB2 phosphorylation and signaling complex formation in carcinoma cells. J Biol Chem 280(10):8875–8883

    Article  CAS  PubMed  Google Scholar 

  • 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):16667–16680

    Article  CAS  PubMed  Google Scholar 

  • 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):1711–1719

    Article  CAS  PubMed  Google Scholar 

  • 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):1758–1769

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • 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):3901–3908

    Article  CAS  PubMed  Google Scholar 

  • 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):1232–1237

    PubMed  Google Scholar 

  • 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):10–30

    Article  PubMed  Google Scholar 

  • 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):245–254

    Article  CAS  PubMed  Google Scholar 

  • 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):2852–2860

    Article  CAS  PubMed  Google Scholar 

  • 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):1141–1145

    CAS  PubMed  Google Scholar 

  • 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):38509–38520

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • 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):1197–1209

    Article  CAS  PubMed  Google Scholar 

  • Lamouille S, Xu J, Derynck R (2014) Molecular mechanisms of epithelial-mesenchymal transition. Nat Rev Mol Cell Biol 15(3):178–196

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • 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):12101–12114

    CAS  PubMed  PubMed Central  Google Scholar 

  • Lokeshwar VB, Mirza S, Jordan A (2014) Targeting hyaluronic acid family for cancer chemoprevention and therapy. Adv Cancer Res 123:35–65

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • 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):114095–114108

    Article  PubMed  PubMed Central  Google Scholar 

  • 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):17618–17630

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • 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):34936–34941

    Article  CAS  PubMed  Google Scholar 

  • 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):1472–1484

    Article  CAS  PubMed  Google Scholar 

  • 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):1507–1517

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • 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 11:418

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • 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 6:3

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Parkin DM (2008) The global burden of urinary bladder cancer. Scand J Urol Nephrol Suppl 218:12–20

    Article  Google Scholar 

  • Ploeg M, Aben KKH, Kiemeney LA (2009) The present and future burden of urinary bladder cancer in the world. World J Urol 27(3):289–293

    Article  PubMed  PubMed Central  Google Scholar 

  • 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):335–342

    Article  CAS  PubMed  Google Scholar 

  • 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):6950–6956

    Article  CAS  PubMed  Google Scholar 

  • Stern R, Asari AA, Sugahara KN (2006) Hyaluronan fragments: an information-rich system. Eur J Cell Biol 85(8):699–715

    Article  CAS  PubMed  Google Scholar 

  • 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):341–346

    Article  CAS  PubMed  Google Scholar 

  • Toole BP (2004) Hyaluronan: from extracellular glue to pericellular cue. Nat Rev Cancer 4(7):528–539

    Article  CAS  PubMed  Google Scholar 

  • Toole BP, Hascall VC (2002) Hyaluronan and tumor growth. Am J Pathol 161(3):745–747

    Article  PubMed  PubMed Central  Google Scholar 

  • Turley EA, Noble PW, Bourguignon LYW (2002) Signaling properties of hyaluronan receptors. J Biol Chem 277(7):4589–4592

    Article  CAS  PubMed  Google Scholar 

  • 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):662–672

    Article  CAS  PubMed  Google Scholar 

  • 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):3534–3541

    PubMed  Google Scholar 

  • 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):324–338

    Article  CAS  Google Scholar 

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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.

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Authors and Affiliations

  1. Department of Biochemistry, All India Institute of Medical Sciences (AIIMS), New Delhi, 110029, India

    Vivek Anand, Madhuram Khandelwal, Sandeep Appunni, Nidhi Gupta & Alpana Sharma

  2. Department of Urology, All India Institute of Medical Sciences (AIIMS), New Delhi, India

    Amlesh Seth & Prabhjot Singh

  3. Department of Pathology, All India Institute of Medical Sciences (AIIMS), New Delhi, India

    Sandeep Mathur

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  1. Vivek Anand
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Anand, V., Khandelwal, M., Appunni, S. et al. CD44 splice variant (CD44v3) promotes progression of urothelial carcinoma of bladder through Akt/ERK/STAT3 pathways: novel therapeutic approach. J Cancer Res Clin Oncol 145, 2649–2661 (2019). https://doi.org/10.1007/s00432-019-03024-9

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