Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

Keynote webinar | Spotlight on medication adherence

LIVE: Thursday 27th June 2024, 18:00-19:30 (CEST)
Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.
ADVANCED SEARCH
Log in
Top
Cancer Chemotherapy and Pharmacology
Published in: Cancer Chemotherapy and Pharmacology 3/2015

01-09-2015 | Original Article

Gossypol sensitizes the antitumor activity of 5-FU through down-regulation of thymidylate synthase in human colon carcinoma cells

Authors: Dan Yang, Jinglei Qu, Xiujuan Qu, Yubo Cao, Ling Xu, Kezuo Hou, Wanyu Feng, Yunpeng Liu

Published in: Cancer Chemotherapy and Pharmacology | Issue 3/2015

Login to get access

Abstract

Purpose

5-Fluorouracil (5-FU) is the basic chemotherapeutic agent used to treat colon cancer. However, the sensitivity of colon cancer cells to 5-FU is limited. Gossypol is a polyphenolic extract of cottonseeds. The purpose of this study was to investigate the activities and related mechanism of gossypol alone or in combination with 5-FU against human colon carcinoma cells.

Methods

The IC50 of gossypol or/and 5-FU in vitro was tested by 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, and the drug interaction was analyzed using the CalcuSyn method. Cell apoptosis was determined using presidium iodide staining and flow cytometric analysis. Western blotting was used to determine the expression of proteins. Transient transfection method was used to silence protein.

Results

The IC50 at 48 h of gossypol in colon cancer cells was 26.11 ± 1.04 μmol/L in HT-29 cells, 14.11 ± 1.08 μmol/L in HCT116 cells, and 21.83 ± 1.05 μmol/L in RKO cells. When gossypol was combined with 5-FU, a synergistic cytotoxic effect was observed in HT-29 cells, HCT116 cells, and RKO cells compared with treatment with gossypol or 5-FU alone. The Western blotting results indicated that gossypol down-regulated thymidylate synthase (TS) rather than thymidine phosphorylase protein expression. Furthermore, the mTOR/p70S6K1 signaling pathway was inhibited in gossypol-treated colon cancer cells, and consequently, cyclin D1 expression was decreased, suggesting an additional mechanism of the observed antiproliferative synergistic interactions. All the observation was confirmed by silencing TS and inactivating the mTOR/p70S6K1 signaling pathway by rapamycin, both of which increased the chemo-sensitizing efficacy of 5-FU.

Conclusions

These findings suggest that gossypol-mediated down-regulation of TS, cyclin D1, and the mTOR/p70S6K1 signaling pathways enhances the anti-tumor effect of 5-FU. Ultimately, our data exposed a new action for gossypol as an enhancer of 5-FU-induced cell growth suppression.
Literature
1.
Likui W, Hong W, Shuwen Z, Yuangang Y, Yan W (2011) The potential of osteopontin as a therapeutic target for human colorectal cancer. J Gastrointest Surg 15(4):652–659CrossRefPubMed
2.
Antonic V, Stojadinovic A, Kester KE, Weina PJ, Brucher BL et al (2013) Significance of infectious agents in colorectal cancer development. J Cancer 4(3):227–240PubMedCentralCrossRefPubMed
3.
Chen J, Lu H, Yan D, Cui F, Wang X, Yu F et al (2015) PAK6 increase chemoresistance and is a prognostic marker for stage II and III colon cancer patients undergoing 5-FU based chemotherapy. Oncotarget 6(1):355–367PubMedCentralPubMed
4.
de la Cruz-Morcillo MA, Valero ML, Callejas-Valera JL, Arias-Gonzalez L, Melgar-Rojas P, Galan-Moya EM et al (2012) P38MAPK is a major determinant of the balance between apoptosis and autophagy triggered by 5-fluorouracil: implication in resistance. Oncogene 31(9):1073–1085CrossRefPubMed
5.
Van Loon K, Venook AP (2011) Adjuvant treatment of colon cancer: what is next? Curr Opin Oncol 23(4):403–409CrossRefPubMed
6.
Soreide K, Berg M, Skudal BS, Nedreboe BS (2011) Advances in the understanding and treatment of colorectal cancer. Discov Med 12(66):393–404PubMed
7.
Shakibaei M, Mobasheri A, Lueders C, Busch F, Shayan P, Goel A (2013) Curcumin enhances the effect of chemotherapy against colorectal cancer cells by inhibition of NF-kappaB and Src protein kinase signaling pathways. PLoS ONE 8(2):e57218PubMedCentralCrossRefPubMed
8.
Yang SY, Miah A, Sales KM, Fuller B, Seifalian AM, Winslet M (2011) Inhibition of the p38 MAPK pathway sensitises human colon cancer cells to 5-fluorouracil treatment. Int J Oncol 38(6):1695–1702PubMed
9.
Cassidy J, Saltz LB, Giantonio BJ, Kabbinavar FF, Hurwitz HI, Rohr UP (2010) Effect of bevacizumab in older patients with metastatic colorectal cancer: pooled analysis of four randomized studies. J Cancer Res Clin Oncol 136(5):737–743PubMedCentralCrossRefPubMed
10.
Pang X, Wu Y, Wu Y, Lu B, Chen J, Wang J et al (2011) (-)-Gossypol suppresses the growth of human prostate cancer xenografts via modulating VEGF signaling-mediated angiogenesis. Mol Cancer Ther 10(5):795–805PubMedCentralCrossRefPubMed
11.
Li H, Piao L, Xu P, Ye W, Zhong S, Lin SH et al (2011) Liposomes containing (−)-gossypol-enriched cottonseed oil suppress Bcl-2 and Bcl-xL expression in breast cancer cells. Pharm Res 28(12):3256–3264CrossRefPubMed
12.
Gunassekaran GR, Priya DK, Gayathri R, Sakthisekaran D (2011) In vitro and in vivo studies on antitumor effects of gossypol on human stomach adenocarcinoma (AGS) cell line and MNNG induced experimental gastric cancer. Biochem Biophys Res Commun 411(4):661–666CrossRefPubMed
13.
Chien CC, Ko CH, Shen SC, Yang LY, Chen YC (2012) The role of COX-2/PGE2 in gossypol-induced apoptosis of colorectal carcinoma cells. J Cell Physiol 227(8):3128–3137CrossRefPubMed
14.
Sadahira K, Sagawa M, Nakazato T, Uchida H, Ikeda Y, Okamoto S et al (2014) Gossypol induces apoptosis in multiple myeloma cells by inhibition of interleukin-6 signaling and Bcl-2/Mcl-1 pathway. Int J Oncol 45(6):2278–2286PubMed
15.
Soderquist RS, Danilov AV, Eastman A (2014) Gossypol increases expression of the pro-apoptotic BH3-only protein NOXA through a novel mechanism involving phospholipase A2, cytoplasmic calcium, and endoplasmic reticulum stress. J Biol Chem 289(23):16190–16199PubMedCentralCrossRefPubMed
16.
Hsiao WT, Tsai MD, Jow GM, Tien LT, Lee YJ (2012) Involvement of Smac, p53, and caspase pathways in induction of apoptosis by gossypol in human retinoblastoma cells. Mol Vis 18:2033–2042PubMedCentralPubMed
17.
Jang GH, Lee M (2014) BH3-mimetic gossypol-induced autophagic cell death in mutant BRAF melanoma cells with high expression of p21Cip1. Life Sci 102(1):41–48CrossRefPubMed
18.
Ahn JH, Jang GH, Lee M (2013) Defective autophagy in multidrug resistant cells may lead to growth inhibition by BH3-mimetic gossypol. J Cell Physiol 228(7):1496–1505CrossRefPubMed
19.
Jiang J, Ye W, Lin YC (2009) Gossypol inhibits the growth of MAT-LyLu prostate cancer cells by modulation of TGFbeta/Akt signaling. Int J Mol Med 24(1):69–75CrossRefPubMed
20.
Cengiz E, Karaca B, Kucukzeybek Y, Gorumlu G, Gul MK, Erten C et al (2010) Overcoming drug resistance in hormone- and drug-refractory prostate cancer cell line, PC-3 by docetaxel and gossypol combination. Mol Biol Rep 37(3):1269–1277CrossRefPubMed
21.
Wong FY, Liem N, Xie C, Yan FL, Wong WC, Wang L et al (2012) Combination therapy with gossypol reveals synergism against gemcitabine resistance in cancer cells with high BCL-2 expression. PLoS ONE 7(12):e50786PubMedCentralCrossRefPubMed
22.
Sung B, Ravindran J, Prasad S, Pandey MK, Aggarwal BB (2010) Gossypol induces death receptor-5 through activation of the ROS-ERK-CHOP pathway and sensitizes colon cancer cells to TRAIL. J Biol Chem 285(46):35418–35427PubMedCentralCrossRefPubMed
23.
Baggstrom MQ, Qi Y, Koczywas M, Argiris A, Johnson EA, Millward MJ et al (2011) A phase II study of AT-101 (gossypol) in chemotherapy-sensitive recurrent extensive-stage small cell lung cancer. J Thorac Oncol 6(10):1757–1760PubMedCentralCrossRefPubMed
24.
Heist RS, Fain J, Chinnasami B, Khan W, Molina JR, Sequist LV et al (2010) Phase I/II study of AT-101 with topotecan in relapsed and refractory small cell lung cancer. J Thorac Oncol 5(10):1637–1643CrossRefPubMed
25.
Formentini A, Henne-Bruns D, Kornmann M (2012) Thymidylate synthase and cyclin D1 protein expression in lymph node negative colorectal cancer: role as prognostic factors? Hepatogastroenterology 59(118):1859–1864PubMed
26.
Sprio AE, Di Scipio F, Ceppi P, Salamone P, Di Carlo F, Scagliotti GV et al (2012) Differentiation-inducing factor-1 enhances 5-fluorouracil action on oral cancer cells inhibiting E2F1 and thymidylate synthase mRNAs accumulation. Cancer Chemother Pharmacol 69(4):983–989CrossRefPubMed
27.
Vinod BS, Antony J, Nair HH, Puliyappadamba VT, Saikia M, Narayanan SS et al (2013) Mechanistic evaluation of the signaling events regulating curcumin-mediated chemosensitization of breast cancer cells to 5-fluorouracil. Cell Death Dis 4:e505PubMedCentralCrossRefPubMed
28.
Ko JC, Tsai MS, Chiu YF, Weng SH, Kuo YH, Lin YW (2011) Up-regulation of extracellular signal-regulated kinase 1/2-dependent thymidylate synthase and thymidine phosphorylase contributes to cisplatin resistance in human non-small-cell lung cancer cells. J Pharmacol Exp Ther 338(1):184–194CrossRefPubMed
29.
Yasuno M, Mori T, Koike M, Takahashi K, Toi M, Takizawa T et al (2005) Importance of thymidine phosphorylase expression in tumor stroma as a prognostic factor in patients with advanced colorectal carcinoma. Oncol Rep 13(3):405–412PubMed
30.
Ye DJ, Zhang JM (2013) Research development of the relationship between thymidine phosphorylase expression and colorectal carcinoma. Cancer Biol Med 10(1):10–15PubMedCentralPubMed
31.
Jastrzebski K, Hannan KM, Tchoubrieva EB, Hannan RD, Pearson RB (2007) Coordinate regulation of ribosome biogenesis and function by the ribosomal protein S6 kinase, a key mediator of mTOR function. Growth Factors 25(4):209–226CrossRefPubMed
32.
Duvel K, Yecies JL, Menon S, Raman P, Lipovsky AI, Souza AL et al (2010) Activation of a metabolic gene regulatory network downstream of mTOR complex 1. Mol Cell 39(2):171–183PubMedCentralCrossRefPubMed
33.
Magnuson B, Ekim B, Fingar DC (2012) Regulation and function of ribosomal protein S6 kinase (S6K) within mTOR signalling networks. Biochem J 441(1):1–21CrossRefPubMed
34.
Hong S, Zhao B, Lombard DB, Fingar DC, Inoki K (2014) Cross-talk between sirtuin and mammalian target of rapamycin complex 1 (mTORC1) signaling in the regulation of S6 kinase 1 (S6K1) phosphorylation. J Biol Chem 289(19):13132–13141PubMedCentralCrossRefPubMed
35.
Kim SJ, Kim JH, Jung HS, Lee TJ, Lee KM, Chang IH (2014) Phosphorylated p70S6 K in noninvasive low-grade urothelial carcinoma of the bladder: correlation with tumor recurrence. Asian J Androl 16(4):611–617PubMedCentralCrossRefPubMed
36.
Li J, Xue L, Hao H, Li R, Luo J (2014) Rapamycin combined with celecoxib enhanced antitumor effects of mono treatment on chronic myelogenous leukemia cells through downregulating mTOR pathway. Tumour Biol 35(7):6467–6474CrossRefPubMed
37.
Axelrod MJ, Gordon V, Mendez RE, Leimgruber SS, Conaway MR, Sharlow ER et al (2014) p70S6 kinase is a critical node that integrates HER-family and PI3 kinase signaling networks. Cell Signal 26(8):1627–1635PubMedCentralCrossRefPubMed
38.
Shigematsu H, Yoshida K, Sanada Y, Osada S, Takahashi T, Wada Y et al (2010) Rapamycin enhances chemotherapy-induced cytotoxicity by inhibiting the expressions of TS and ERK in gastric cancer cells. Int J Cancer 126(11):2716–2725PubMed
39.
Deng J, Lei W, Fu JC, Zhang L, Li JH, Xiong JP (2014) Targeting miR-21 enhances the sensitivity of human colon cancer HT-29 cells to chemoradiotherapy in vitro. Biochem Biophys Res Commun 443(3):789–795CrossRefPubMed
40.
Johnson PW (2008) New targets for lymphoma treatment. Ann Oncol 19(Suppl 4):iv56–iv59PubMed
41.
Kim KW, Kwon HC, Kim SH, Oh SY, Lee S, Lee JH et al (2013) Prognostic significance of thymidylate synthase, thymidine phosphorylase and dihydropyrimidine dehydrogenase expression in biliary tract cancer patients receiving adjuvant 5-fluorouracil-based chemotherapy. Mol Clin Oncol 1(6):987–994PubMedCentralPubMed
42.
Rose MG, Farrell MP, Schmitz JC (2002) Thymidylate synthase: a critical target for cancer chemotherapy. Clin Colorectal Cancer 1(4):220–229CrossRefPubMed
43.
Di Cresce C, Figueredo R, Ferguson PJ, Vincent MD, Koropatnick J (2011) Combining small interfering RNAs targeting thymidylate synthase and thymidine kinase 1 or 2 sensitizes human tumor cells to 5-fluorodeoxyuridine and pemetrexed. J Pharmacol Exp Ther 338(3):952–963CrossRefPubMed
44.
Bhattacharya B, Akram M, Balasubramanian I, Tam KK, Koh KX, Yee MQ et al (2012) Pharmacologic synergy between dual phosphoinositide-3-kinase and mammalian target of rapamycin inhibition and 5-fluorouracil in PIK3CA mutant gastric cancer cells. Cancer Biol Ther 13(1):34–42CrossRefPubMed
45.
Liao F, Yang Z, Lu X, Guo X, Dong W (2013) Sinomenine sensitizes gastric cancer cells to 5-fluorouracil in vitro and in vivo. Oncol Lett 6(6):1604–1610PubMedCentralPubMed
46.
Sprio AE, Di Scipio F, Ceppi P, Salamone P, Di Carlo F, Scagliotti GV et al (2012) Differentiation-inducing factor-1 enhances 5-fluorouracil action on oral cancer cells inhibiting E2F1 and thymidylate synthase mRNAs accumulation. Cancer Chemother Pharmacol 69(4):983–989CrossRefPubMed
47.
Di Gennaro E, Bruzzese F, Pepe S, Leone A, Delrio P, Subbarayan PR et al (2009) Modulation of thymidilate synthase and p53 expression by HDAC inhibitor vorinostat resulted in synergistic antitumor effect in combination with 5FU or raltitrexed. Cancer Biol Ther 8(9):782–791CrossRefPubMed
48.
Jin HS, Lee DH, Kim DH, Chung JH, Lee SJ, Lee TH (2009) cIAP1, cIAP2, and XIAP act cooperatively via nonredundant pathways to regulate genotoxic stress-induced nuclear factor-kappaB activation. Cancer Res 69(5):1782–1791CrossRefPubMed
49.
Murray PE, McNally VA, Lockyer SD, Williams KJ, Stratford IJ, Jaffar M et al (2002) Synthesis and enzymatic evaluation of pyridinium-substituted uracil derivatives as novel inhibitors of thymidine phosphorylase. Bioorg Med Chem 10(3):525–530CrossRefPubMed
50.
Evrard A, Cuq P, Robert B, Vian L, Pelegrin A, Cano JP (1999) Enhancement of 5-fluorouracil cytotoxicity by human thymidine-phosphorylase expression in cancer cells: in vitro and in vivo study. Int J Cancer 80(3):465–470CrossRefPubMed
51.
Evrard A, Cuq P, Ciccolini J, Vian L, Cano JP (1999) Increased cytotoxicity and bystander effect of 5-fluorouracil and 5-deoxy-5-fluorouridine in human colorectal cancer cells transfected with thymidine phosphorylase. Br J Cancer 80(11):1726–1733PubMedCentralCrossRefPubMed
52.
Seeliger H, Guba M, Koehl GE, Doenecke A, Steinbauer M, Bruns CJ et al (2004) Blockage of 2-deoxy-d-ribose-induced angiogenesis with rapamycin counteracts a thymidine phosphorylase-based escape mechanism available for colon cancer under 5-fluorouracil therapy. Clin Cancer Res 10(5):1843–1852CrossRefPubMed
53.
Nagaraju GP, Alese OB, Landry J, Diaz R, El-Rayes BF (2014) HSP90 inhibition downregulates thymidylate synthase and sensitizes colorectal cancer cell lines to the effect of 5FU-based chemotherapy. Oncotarget 5(20):9980–9991PubMedCentralPubMed
54.
Ohtani K (1999) Implication of transcription factor E2F in regulation of DNA replication. Front Biosci 4:D793–D804CrossRefPubMed
55.
Advani SH (2010) Targeting mTOR pathway: a new concept in cancer therapy. Indian J Med Paediatr Oncology 31(4):132–136CrossRef
56.
Shuai XM, Han GX, Wang GB, Chen JH (2006) Cyclin D1 antisense oligodexoyneucleotides inhibits growth and enhances chemosensitivity in gastric carcinoma cells. World J Gastroenterol 12(11):1766–1769PubMedCentralPubMed
57.
Kornmann M, Danenberg KD, Arber N, Beger HG, Danenberg PV, Korc M (1999) Inhibition of cyclin D1 expression in human pancreatic cancer cells is associated with increased chemosensitivity and decreased expression of multiple chemoresistance genes. Cancer Res 59(14):3505–3511PubMed
58.
Wullschleger S, Loewith R, Hall MN (2006) TOR signaling in growth and metabolism. Cell 124(3):471–484CrossRefPubMed
59.
Bhattacharya B, Akram M, Balasubramanian I, Tam KK, Koh KX, Yee MQ et al (2012) Pharmacologic synergy between dual phosphoinositide-3-kinase and mammalian target of rapamycin inhibition and 5-fluorouracil in PIK3CA mutant gastric cancer cells. Cancer Biol Ther 13(1):34–42CrossRefPubMed
60.
Tomioka H, Mukohara T, Kataoka Y, Ekyalongo RC, Funakoshi Y, Imai Y et al (2012) Inhibition of the mTOR/S6 K signal is necessary to enhance fluorouracil-induced apoptosis in gastric cancer cells with HER2 amplification. Int J Oncol 41(2):551–558PubMed
61.
Atreya CE, Ducker GS, Feldman ME, Bergsland EK, Warren RS, Shokat KM (2012) Combination of ATP-competitive mammalian target of rapamycin inhibitors with standard chemotherapy for colorectal cancer. Invest New Drugs 30(6):2219–2225CrossRefPubMed
62.
O’Reilly T, McSheehy PM, Wartmann M, Lassota P, Brandt R, Lane HA (2011) Evaluation of the mTOR inhibitor, everolimus, in combination with cytotoxic antitumor agents using human tumor models in vitro and in vivo. Anticancer Drugs 22(1):58–78CrossRefPubMed
63.
Lee KH, Hur HS, Im SA, Lee J, Kim HP, Yoon YK et al (2010) RAD001 shows activity against gastric cancer cells and overcomes 5-FU resistance by downregulating thymidylate synthase. Cancer Lett 299(1):22–28CrossRefPubMed
64.
Hosono Y, Osada S, Nawa M, Takahashi T, Yamaguchi K, Kawaguchi Y et al (2010) Combination therapy of 5-fluorouracil with rapamycin for hormone receptor-negative human breast cancer. Anticancer Res 30(7):2625–2630PubMed
65.
Dal Col J, Dolcetti R (2008) GSK-3beta inhibition: at the crossroad between Akt and mTOR constitutive activation to enhance cyclin D1 protein stability in mantle cell lymphoma. Cell Cycle 7(18):2813–2816CrossRefPubMed
66.
Grzybowska-Izydorczyk O, Smolewski P (2012) mTOR kinase inhibitors as a treatment strategy in hematological malignancies. Future Med Chem 4(4):487–504CrossRefPubMed
67.
Cheng P, Ni Z, Dai X, Wang B, Ding W, Rae Smith A et al (2013) The novel BH-3 mimetic apogossypolone induces Beclin-1- and ROS-mediated autophagy in human hepatocellular carcinoma cells. Cell Death Dis 4:e489PubMedCentralCrossRefPubMed
Metadata
Title
Gossypol sensitizes the antitumor activity of 5-FU through down-regulation of thymidylate synthase in human colon carcinoma cells
Authors
Dan Yang
Jinglei Qu
Xiujuan Qu
Yubo Cao
Ling Xu
Kezuo Hou
Wanyu Feng
Yunpeng Liu
Publication date
01-09-2015
Publisher
Springer Berlin Heidelberg
Published in
Cancer Chemotherapy and Pharmacology / Issue 3/2015
Print ISSN: 0344-5704
Electronic ISSN: 1432-0843
DOI
https://doi.org/10.1007/s00280-015-2749-0

Other articles of this Issue 3/2015

Cancer Chemotherapy and Pharmacology 3/2015 Go to the issue

Original Article

Modeling and simulations relating overall survival to tumor growth inhibition in renal cell carcinoma patients

Original Article

Phase I study of combination of pasireotide LAR + gemcitabine in locally advanced or metastatic pancreatic cancer

Original Article

Integrated nonclinical and clinical risk assessment of the investigational proteasome inhibitor ixazomib on the QTc interval in cancer patients

Original Article

A phase II study of icotinib and whole-brain radiotherapy in Chinese patients with brain metastases from non-small cell lung cancer

Original Article

Characterization of the metabolism of benzaldehyde dimethane sulfonate (NSC 281612, DMS612)

Original Article

AT-101 inhibits hedgehog pathway activity and cancer growth
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
Image Credits