Skip to main content
SpringerLink
Log in

Sodium butyrate induces cell death by autophagy and reactivates a tumor suppressor gene DIRAS1 in renal cell carcinoma cell line UOK146

  • Published:
In Vitro Cellular & Developmental Biology - Animal Aims and scope Submit manuscript

Abstract

Sodium butyrate (SB), a histone deacetylase inhibitor, is emerging as a potent anti-cancer drug for different types of cancers. In the present study, anti-cancer activity of SB in Xp11.2 (TFE3) translocated renal cell carcinoma cell line UOK146 was studied. Anti-proliferative effect of SB in renal cell carcinoma (RCC) cell line UOK146 was evaluated by MTT assay and morphological characteristics were observed by phase contrast microscopy which displayed the cell death after SB treatment. SB induces DNA fragmentation and change in nuclear morphology observed by increased sub-G1 region cell population and nuclear blebbings. Cell cycle arrest at G2/M phase was found after SB treatment. UOK146 cell line shows autophagy mode of cell death as displayed by acridine orange staining and flow cytometry analysis. LC3-II, a protein marker of autophagy, was also found to be upregulated after SB treatment. A tumor suppressor gene DIRAS1 was upregulated after SB treatment, displaying its anti-cancer potential at molecular level. These findings suggest that SB could serve as a novel regulator of tumor suppressors and lead to the discovery of novel therapeutics with better and enhanced anti-cancer activity.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Abramova MV, Pospelova TV, Nikulenkov FP, Hollander CM, Fornace AJ Jr, Pospelov VA (2006) G1/S arrest induced by histone deacetylase inhibitor sodium butyrate in E1A + Ras-transformed cells is mediated through down-regulation of E2F activity and stabilization of beta-catenin. J Biol Chem 281:21040–21051

    Article  CAS  PubMed  Google Scholar 

  • Archer SY, Hodin RA (1999) Histone acetylation and cancer. Curr Opin Genet Dev 9:171–174

    Article  CAS  PubMed  Google Scholar 

  • Bai Z, Zhang Z, Ye Y, Wang S (2010) Sodium butyrate induces differentiation of gastric cancer cells to intestinal cells via the PTEN/phosphoinositide 3-kinase pathway. Cell Biol Int 34:1141–1145

    Article  CAS  PubMed  Google Scholar 

  • Bergom C, Hauser AD, Rymaszewski A, Gonyo P, Prokop JW, Jennings BC, Lawton AJ, Frei A, Lorimer EL, Aguilera-Barrantes I, Mackinnon AC, Noon K, Fierke CA, Williams CL (2016) The tumor-suppressive small GTPase DiRas1 binds the noncanonical guanine nucleotide exchange factor SmgGDS and antagonizes SmgGDS interactions with oncogenic small GTPases. J Biol Chem 291:6534–6545

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Cianfanelli V, Fuoco C, Lorente M, Salazar M, Quondamatteo F, Gherardini PF et al (2015) AMBRA1 links autophagy to cell proliferation and tumorigenesis by promoting c-Myc dephosphorylation and degradation. Nat Cell Biol 17:20–30. https://doi.org/10.1038/ncb3072

    Article  CAS  PubMed  Google Scholar 

  • D’Argenio G, Mazzacca G (1999) Short-chain fatty acid in the human colon. Relation to inflammatory bowel diseases and colon cancer. Adv Exp Med Biol 472:149–158

    Article  PubMed  Google Scholar 

  • Dadakhujaev S, Noh HS, Jung EJ, Hah YS, Kim CJ, Kim DR (2008) The reduced catalase expression in TrkA-induced cells leads to autophagic cell death via ROS accumulation. Exp Cell Res 314:3094–3106

    Article  CAS  Google Scholar 

  • Damaskos C, Karatzas T, Nikolidakis L, Kostakis ID, Karamaroudis S, Boutsikos G, Damaskou Z, Kostakis A, Kouraklis G (2015) Histone deacetylase (HDAC) inhibitors: current evidence for therapeutic activities in pancreatic cancer. Anticancer Res 35:3129–3135

    CAS  PubMed  Google Scholar 

  • Dos Santos MP, de Farias CB, Roesler R, Brunetto AL, Abujamra AL (2014) In vitro antitumor effect of sodium butyrate and zoledronic acid combined with traditional chemotherapeutic drugs: a paradigm of synergistic molecular targeting in the treatment of Ewing sarcoma. Oncol Rep 31:955–968

    Article  PubMed  Google Scholar 

  • Elizabeth A, Williams Jonathan M, Coxhead and John C Mathers (2003) Anti-cancer effects of butyrate: use of microarray technologies to investigate mechanisms. Proc Nutr Soc 62:107–115

  • Gotfryd K, Skladchikova G, Lepekhin EA, Berezin V, Bock E, Walmod PS (2010) Cell type-specific anti-cancer properties of valproic acid: independent effects on HDAC activity and Erk1/2 phosphorylation. BMC Cancer 10:383

    Article  PubMed  PubMed Central  Google Scholar 

  • Gozuacik D, Kimchi A (2004) Autophagy as a cell death and tumor suppressor mechanism. Oncogene 23(16):2891–2906

    Article  CAS  PubMed  Google Scholar 

  • Kanzawa T, Kondo Y, Ito H, Kondo S, Germano I (2003) Induction of autophagic cell death in malignant glioma cells by arsenic trioxide. Cancer Res 63:2103–2108

    CAS  PubMed  Google Scholar 

  • Kitazono M, Shinchi H, Ishigami S, Ueno S, Natsugoe S (2010) Effects of histone deacetylase inhibitor, sodium butyrate, on 53-kDa protein expression and sensitivity to anti-cancer drugs of pancreatic cancer cells. Curr Ther Res 71:162–172

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kurz EU, Wilson SE, Leader KB, Sampey BP, Allan WP, Yalowich JC, Kroll DJ (2001) The histone deacetylase inhibitor sodium butyrate induces DNA topoisomerase II alpha expression and confers hypersensitivity to etoposide in human leukemic cell lines. Mol Cancer Ther 1:121–131

    CAS  PubMed  Google Scholar 

  • Marchion D, Münster P (2007) Development of histone deacetylase inhibitors for cancer treatment. Expert Rev Anticancer Ther 7:583–598

    Article  CAS  PubMed  Google Scholar 

  • Mathiassen SG, De Zio D, Cecconi F (2017) Autophagy and the cell cycle: a complex landscape. Front Oncol 7:51

    Article  PubMed  PubMed Central  Google Scholar 

  • Mu D, Gao Z, Guo H, Zhou G, Sun B (2013) Sodium butyrate induces growth inhibition and apoptosis in human prostate cancer DU145 cells by up-regulation of the expression of annexin A1. PLoS One 8:e74922

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ogawa H, Rafiee P, Fisher PJ, Johnson NA, Otterson MF, Binion DG, Ogawa H, Rafiee P, Fisher PJ, Johnson NA, Otterson MF, Binion DG (2003) Sodium butyrate inhibits angiogenesis of human intestinal microvascular endothelial cells through COX-2 inhibition. FEBS Lett 554:88–94

    Article  CAS  PubMed  Google Scholar 

  • Papi A, Ferreri AM, Rocchi P, Guerra F, Orlandi M (2010) Epigenetic modifiers as anticancer drugs: effectiveness of valproic acid in neural crest-derived tumor cells. Anticancer Res 30:535–540

    CAS  PubMed  Google Scholar 

  • Parikh J, Coleman T, Messias N, Brown J (2009) Temsirolimus in the treatment of renal cell carcinoma associated with Xp11.2 translocation/TFE gene fusion proteins: a case report and review of literature. Rare Tumors 1:e53

  • Pwint TP, Macaulay V, Roberts IS, Sullivan M, Protheroe A (2011) An adult Xp11.2 translocation renal carcinoma showing response to treatment with sunitinib. Urol Oncol 29:821–824

    Article  CAS  PubMed  Google Scholar 

  • Ropero S, Esteller M (2007) The role of histone deacetylases (HDACs) in human cancer. Mol Oncol 1:19–25

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rosato RR, Almenara JA, Yu C, Grant S (2004) Evidence of a functional role for p21WAF1/CIP1 downregulation in synergistic antileukemic interactions between the histone deacetylase inhibitor sodium butyrate and flavopiridol. Mol Pharmacol 65:571–581

    Article  CAS  PubMed  Google Scholar 

  • Säemann MD, Böhmig GA, Osterreicher CH, Burtscher H, Parolini O, Diakos C, Stöckl J, Hörl WH, Zlabinger GJ (2000) Anti-inflammatory effects of sodium butyrate on human monocytes: potent inhibition of IL-12 and up-regulation of IL-10 production. FASEB J 14:2380–2382

    Article  PubMed  Google Scholar 

  • Sato H, Uzu M, Kashiba T, Suzuki R, Fujiwara T, Okuzawa H, Ueno K (2017) Sodium butyrate enhances the growth inhibitory effect of sunitinib in human renal cell carcinoma cells. Oncol Lett 14:937–943

    Article  PubMed  PubMed Central  Google Scholar 

  • Sidhar SK, Clark J, Gill S, Hamoudi R, Crew AJ, Gwilliam R, Ross M, Linehan WM, Birdsall S, Shipley J, Cooper CS (1996) The t(X;1)(p11.2;q21.2) translocation in papillary renal cell carcinoma fuses a novel gene PRCC to the TFE3 transcription factor gene. Hum Mol Genet 5:1333–1338

    Article  CAS  PubMed  Google Scholar 

  • Sutton MN, Lu Z, Bast RC Jr. (2016) DIRAS1 and DIRAS2 are tumor suppressor candidates that inhibit cell growth, motility, and proliferation while regulating autophagy in ovarian cancer. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research: Exploiting Vulnerabilities; Oct 17–20, 2015; Orlando, FL. Philadelphia (PA): AACR; Clin Cancer Res. 22(2 Suppl):Abstract nr A43

  • Verma SP, Sisoudiya S, Das P (2015) Aqueous extract of anticancer drug CRUEL herbomineral formulation capsules exerts anti-proliferative effects in renal cell carcinoma cell lines. Asian Pac J Cancer Prev 16:8419–8423

    Article  PubMed  Google Scholar 

  • Verma SP, Tripathi VC, Das P (2014) Asparagus racemosus leaf extract inhibits growth of UOK 146 renal cell carcinoma cell line: simultaneous oncogenic PRCCTFE3 fusion transcript inhibition and apoptosis independent cell death. Asian Pac J Cancer Prev 15:1937–1941

    Article  PubMed  Google Scholar 

  • Wang HG, Huang XD, Shen P, Li LR, Xue HT, Ji GZ (2013) Anticancer effects of sodium butyrate on hepatocellular carcinoma cells in vitro. Int J Mol Med 31:967–974

    Article  CAS  PubMed  Google Scholar 

  • Zhang J, Yi M, Zha L, Chen S, Li Z, Li C, Gong M, Deng H, Chu X, Chen J, Zhang Z, Mao L, Sun S (2016) Sodium butyrate induces endoplasmic reticulum stress and autophagy in colorectal cells: implications for apoptosis. PLoS One 11:e0147218

    Article  PubMed  PubMed Central  Google Scholar 

  • Zhu YH, Fu L, Chen L, Qin YR, Liu H, Xie F, Zeng T, Dong SS, Li J, Li Y, Dai Y, Xie D, Guan XY (2013) Downregulation of the novel tumor suppressor DIRAS1 predicts poor prognosis in esophageal squamous cell carcinoma. Cancer Res 73:2298–2309

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We acknowledge W M Linehan, National Cancer Institute, USA, for providing the UOK146 cell line. DBT-BHU Interdisciplinary School of Life Sciences (ISLS) Banaras Hindu University, Varanasi, India, for Flow Cytometry and Real Time PCR facilities. Prof. Prasenjit Guchhait, Regional Centre for Biotechnology, Faridabad, India, for anti-LC3 antibody. The Indian Council of Medical Research (ICMR), Government of India, New Delhi, for fellowship support of JRF and SRF to Shiv Prakash Verma.

Author information

Authors and Affiliations

  1. Centre for Genetic Disorders, Institute of Science, Banaras Hindu University, Varanasi, 221005, India

    Shiv Prakash Verma & Parimal Das

  2. VIT University, Vellore, Tamil Nadu, India

    Ayushi Agarwal

Authors
  1. Shiv Prakash Verma
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ayushi Agarwal
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Parimal Das
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Parimal Das.

Ethics declarations

Competing interests

The authors declare that they have no competing interests.

Additional information

Editor: Tetsuji Okamoto

Appendix

Appendix

GAPDH-Forward: 5′-AGGGCTGCTTTTAACTCTGGT-3′

GAPDH-Reverse: 5′-CCCCACTTGATTTTGGAGGGA-3′

DIRAS1-Forward: 5′-GCAACAAGTGCGATGAGACG-3′

DIRAS1-Reverse: 5′-TCCTCTTCTGCTTCCCGGAG-3’

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Verma, S.P., Agarwal, A. & Das, P. Sodium butyrate induces cell death by autophagy and reactivates a tumor suppressor gene DIRAS1 in renal cell carcinoma cell line UOK146. In Vitro Cell.Dev.Biol.-Animal 54, 295–303 (2018). https://doi.org/10.1007/s11626-018-0239-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11626-018-0239-5

Keywords

Search

Navigation