Abstract
Purpose
HepaCAM, an N-linked glycoprotein that encodes a member of the immunoglobulin superfamily, has been reported to be a tumor suppressor gene that mediates diverse cellular bio-functions. Recent studies have shown that the FoxO transcription factors play a pivotal role during cancer progression. Here, we explored the correlation between HepaCAM and the FoxO family via regulation of the PI3K/AKT pathway.
Methods
HepaCAM and FoxO3 expression were detected by immunohistochemistry staining. We detected the effect of HepaCAM on the proliferation and viability of bladder cancer through AKT signaling by colony formation, the MTT assay and Western blotting. We observed the nuclear translocation of FoxO3 by immunofluorescence staining after expressing HepaCAM.
Results
HepaCAM depletion was discovered in bladder cancer tissues compared with adjacent normal tissues, and the decreased level was associated with the degradation of FoxO3. Furthermore, re-expression of HepaCAM significantly disrupted T24 and BIU-87 cell colony formation, as well as reduced p-AKT and p-FoxO protein expression. We found that the combined treatment of HepaCAM-overexpressing adenovirus with the PI3K inhibitor LY294002 enhanced the inhibitory effects on cell proliferation, viability and protein expression. Additionally, overexpressed HepaCAM decreased the activated effect on cell proliferation, viability and protein expression of the AKT activator SC79. Moreover, we observed that HepaCAM induced nuclear translocation of FoxO3.
Conclusions
Our research implicated that HepaCAM may function as a novel therapeutic target that inhibits the proliferation of bladder cancer via the AKT/FoxO pathway.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Balcazar Morales N, Aguilar de Plata C (2012) Role of AKT/mTORC1 pathway in pancreatic beta-cell proliferation. Colomb Med 43:235–243
Bartell SM et al (2014) FoxO proteins restrain osteoclastogenesis and bone resorption by attenuating H2O2 accumulation. Nat Commun 5:3773 doi:10.1038/ncomms4773
Carbajo-Pescador S, Mauriz JL, Garcia-Palomo A, Gonzalez-Gallego J (2014) FoxO proteins: regulation and molecular targets in liver cancer. Curr Med Chem 21:1231–1246
Chung Moh M, Hoon Lee L, Shen S (2005) Cloning and characterization of hepaCAM, a novel Ig-like cell adhesion molecule suppressed in human hepatocellular carcinoma. J Hepatol 42:833–841. doi:10.1016/j.jhep.2005.01.025
Cohen S, Lee D, Zhai B, Gygi SP, Goldberg AL (2014) Trim32 reduces PI3K-Akt-FoxO signaling in muscle atrophy by promoting plakoglobin-PI3K dissociation. J Cell Biol 204:747–758. doi:10.1083/jcb.201304167
Fritz RD, Varga Z, Radziwill G (2010) CNK1 is a novel Akt interaction partner that promotes cell proliferation through the Akt-FoxO signalling axis. Oncogene 29:3575–3582. doi:10.1038/onc.2010.104
Fruman DA, Rommel C (2014) PI3K and cancer: lessons, challenges and opportunities. Nat Rev Drug Discov 13:140–156. doi:10.1038/nrd4204
Furuyama T, Nakazawa T, Nakano I, Mori N (2000) Identification of the differential distribution patterns of mRNAs and consensus binding sequences for mouse DAF-16 homologues. Biochem J 349:629–634
Greer EL, Brunet A (2008) FOXO transcription factors in ageing and cancer. Acta physiologica 192:19–28. doi:10.1111/j.1748-1716.2007.01780.x
Hawley SA, Ross FA, Gowans GJ, Tibarewal P, Leslie NR, Hardie DG (2014) Phosphorylation by Akt within the ST loop of AMPK-alpha1 down-regulates its activation in tumour cells. Biochem J 459:275–287. doi:10.1042/BJ20131344
Huang H, Tindall DJ (2011) Regulation of FOXO protein stability via ubiquitination and proteasome degradation. Biochim Biophys Acta 1813:1961–1964. doi:10.1016/j.bbamcr.2011.01.007
I OS et al (2015) FoxO1 integrates direct and indirect effects of insulin on hepatic glucose production and glucose utilization. Nat Commun 6:7079. doi:10.1038/ncomms8079
Lin A et al (2014) The FoxO-BNIP3 axis exerts a unique regulation of mTORC1 and cell survival under energy stress. Oncogene 33:3183–3194. doi:10.1038/onc.2013.273
Lv Y, Song S, Zhang K, Gao H, Ma R (2013) CHIP regulates AKT/FoxO/Bim signaling in MCF7 and MCF10A cells. PloS one 8:e83312. doi:10.1371/journal.pone.0083312
Martini M, De Santis MC, Braccini L, Gulluni F, Hirsch E (2014) PI3K/AKT signaling pathway and cancer: an updated review. Ann Med 46:372–383. doi:10.3109/07853890.2014.912836
Moh MC, Zhang C, Luo C, Lee LH, Shen S (2005) Structural and functional analyses of a novel ig-like cell adhesion molecule, hepaCAM, in the human breast carcinoma MCF7 cells. J Biol Chem 280:27366–27374. doi:10.1074/jbc.M500852200
Moh MC, Zhang T, Lee LH, Shen S (2008) Expression of hepaCAM is downregulated in cancers and induces senescence-like growth arrest via a p53/p21-dependent pathway in human breast cancer cells. Carcinogenesis 29:2298–2305. doi:10.1093/carcin/bgn226
Pellicano F et al (2014) The antiproliferative activity of kinase inhibitors in chronic myeloid leukemia cells is mediated by FOXO transcription factors. Stem Cells 32:2324–2337. doi:10.1002/stem.1748
Pramanik KC, Fofaria NM, Gupta P, Srivastava SK (2014) CBP-mediated FOXO-1 acetylation inhibits pancreatic tumor growth by targeting SirT. Mol Cancer Ther 13:687–698. doi:10.1158/1535-7163.MCT-13-0863
Roy SK, Chen Q, Fu J, Shankar S, Srivastava RK (2011) Resveratrol inhibits growth of orthotopic pancreatic tumors through activation of FOXO transcription factors. PloS One 6:e25166. doi:10.1371/journal.pone.0025166
Shiota M et al (2010) Foxo3a suppression of urothelial cancer invasiveness through Twist1, Y-box-binding protein 1, and E-cadherin regulation Clinical cancer research : an official journal of the American Association for. Cancer Res 16:5654–5663. doi:10.1158/1078-0432.CCR-10-0376
Siegel RL, Miller KD, Jemal A (2015) Cancer statistics, 2015. CA Cancer J Clin 65:5–29. doi:10.3322/caac.21254
Song X et al (2014) Overexpression of HepaCAM inhibits cell viability and motility through suppressing nucleus translocation of androgen receptor and ERK signaling in prostate cancer. Prostate 74:1023–1033. doi:10.1002/pros.22817
Tan B et al (2014) HepaCAM inhibits clear cell renal carcinoma 786-0 cell proliferation via blocking PKCepsilon translocation from cytoplasm to plasma membrane. Mol Cell Biochem 391:95–102. doi:10.1007/s11010-014-1991-9
Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal A (2015) Global cancer statistics, 2012. CA Cancer J Clin 65:87–108. doi:10.3322/caac.21262
Wang Q, Luo C, Wu X, Du H, Song X, Fan Y (2013) hepaCAM and p-mTOR closely correlate in bladder transitional cell carcinoma and hepaCAM expression inhibits proliferation via an AMPK/mTOR dependent pathway in human bladder cancer cells. J Urol 190:1912–1918. doi:10.1016/j.juro.2013.05.013
Weigel D, Jurgens G, Kuttner F, Seifert E, Jackle H (1989) The homeotic gene fork head encodes a nuclear protein and is expressed in the terminal regions of the Drosophila embryo. Cell 57:645–658
Xu B, He Y, Wu X, Luo C, Liu A, Zhang J (2012) Exploration of the correlations between interferon-gamma in patient serum and HEPACAM in bladder transitional cell carcinoma, and the interferon-gamma mechanism inhibiting BIU-87 proliferation. J Urol 188:1346–1353. doi:10.1016/j.juro.2012.06.005
Yaklichkin S, Vekker A, Stayrook S, Lewis M, Kessler DS (2007) Prevalence of the EH1 Groucho interaction motif in the metazoan Fox family of transcriptional regulators. BMC Genom 8:201. doi:10.1186/1471-2164-8-201
Zareen N, Biswas SC, Greene LA (2013) A feed-forward loop involving Trib3, Akt and FoxO mediates death of NGF-deprived neurons. Cell Death Differ 20:1719–1730. doi:10.1038/cdd.2013.128
Zhang QL et al (2011a) HepaCAM induces G1 phase arrest and promotes c-Myc degradation in human renal cell carcinoma. J cell biochem 112:2910–2919. doi:10.1002/jcb.23207
Zhang X, Tang N, Hadden TJ, Rishi AK (2011b) Akt, FoxO and regulation of apoptosis. Biochim Biophys Acta 1813:1978–1986. doi:10.1016/j.bbamcr.2011.03.010
Zhang J et al (2015) Histone deacetylase inhibitors induce autophagy through FOXO1-dependent pathways. Autophagy 11:629–642. doi:10.1080/15548627.2015.1023981
Acknowledgements
We thank the patients and their families who generously donated valuable tissue samples.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no conflict of interest.
Funding
This study was supported by grants from the National Natural Science Foundation of China (NSFC) (Grant No. 81072086).
Ethical approval
All of the procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. This study was approved by the Ethics Committee of Chongqing Medical University.
Informed consent
Informed consent was obtained from all individual participants included in the study.
Rights and permissions
About this article
Cite this article
Tang, M., Zhao, Y., Liu, N. et al. Overexpression of HepaCAM inhibits bladder cancer cell proliferation and viability through the AKT/FoxO pathway. J Cancer Res Clin Oncol 143, 793–805 (2017). https://doi.org/10.1007/s00432-016-2333-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00432-016-2333-y