Open Access

Fatty acids inhibit anticancer effects of 5-fluorouracil in mouse cancer cell lines

  • Authors:
    • Eriko Tanabe
    • Misaho Kitayoshi
    • Kiyomu Fujii
    • Hitoshi Ohmori
    • Yi Luo
    • Yui Kadochi
    • Shiori Mori
    • Rina Fujiwara
    • Yukiko Nishiguchi
    • Takamitsu Sasaki
    • Hiroki Kuniyasu
  • View Affiliations

  • Published online on: May 17, 2017     https://doi.org/10.3892/ol.2017.6190
  • Pages: 681-686
  • Copyright: © Tanabe et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

The present study investigated the effects of two major dietary fatty acid components, linoleic acid (LA) and elaidic acid (EA), on the antitumor effects of 5-fluorouracil (5‑FU) in the LL2, CT26 and CMT93 mouse cancer cell lines. Concurrent treatment with LA and 5‑FU elicited a decreased cell viability compared with treatment with 5‑FU alone. In addition, increased inhibition of growth was observed following concurrent treatment with EA and 5‑FU. Sequential treatment of LA followed by 5‑FU abrogated the anticancer effects of 5‑FU, and treatment with EA followed by 5‑FU increased cancer cell growth in addition to abrogating the anticancer effects of 5‑FU. The expression of the stem cell markers CD133 and nucleostemin (NS) increased in all three cell lines treated concurrently with 5‑FU and either LA or EA when compared with cells treated with 5‑FU alone. Aldehyde dehydrogenase activity in the cancer stem cells (CSCs), in response to concurrent treatment with 5‑FU and either LA or EA, was increased compared with 5‑FU treatment alone. 5‑FU inhibited the growth of CT26 tumors, but co‑treatment with either LA or EA abrogated this effect. NS‑positive CSCs were more abundant in CT26 tumors treated with 5‑FU and either LA or EA compared with those treated with 5‑FU alone. The results of the present study suggested that, rather than altering the sensitivity of cancer cells to 5‑FU, LA and EA may promote the survival of CSCs. The results indicated that dietary composition during chemotherapy is an important issue.

Introduction

High intake of fatty food is associated with an increased risk of cancer, particularly for colorectal cancer (CRC) (13). A high-fat diet is associated with aggressive prostate cancer and n-6 fatty acids, including linoleic acid (LA), enhance breast cancer invasion and metastasis (4). LA in combination with azoxymethane results in the upregulation of insulin-like growth factor-I/II receptors (5), Ras (6), cyclooxygenase-2 (7) and high mobility group box-1 (8), which act to accelerate colon carcinogenesis (6,7,9,10). Prolonged treatment with LA induces a quiescent state in CRC cells and the dormancy of subcutaneous tumors in mice, which is thought to be associated with low proliferation of cancer stem cells (CSCs) (11,12).

Trans fatty acids (TFAs) are reported to increase the risk of certain types of cancers, including CRC and prostate cancer (13,14). TFAs, in particular elaidic acid (EA), enhance cancer cell growth, invasion, survival and metastasis through activation of the Wnt signaling pathway and the induction of epithelial-mesenchymal transition (1517). TFAs were previously recognized as safe for human consumption; however, the Food and Drug Administration has proposed that TFAs must be removed from prepared foods by June 2018 (18).

LA and EA are abundant dietary fatty acids, and are part of a normal diet eaten by patients with cancer receiving chemotherapy. The present study focused on the effect of LA and EA on the action of 5-fluorouracil (5-FU), a common anticancer agent, with a particular emphasis on cancer stemness.

Materials and methods

Cell culture

The mouse colon cancer CT26 cell line was provided by Professor Isaiah J. Fidler (MD Anderson Cancer Center, Houston, TX, USA). The mouse rectal carcinoma CMT93 cell line and the mouse lung cancer LL2 cell line were purchased from DS Pharma Biomedical Co., Ltd. (Osaka, Japan). Cells were cultured in Dulbecco's modified Eagle's medium (Wako Pure Chemical Industries, Ltd., Osaka, Japan) supplemented with 10% fetal bovine serum (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) at 37°C in a humidified condition with 5% CO2 atmosphere.

Cells in 6 dishes (1×105 in 3.5 cm-diameter dishes) were treated with LA (50 µg/ml) or EA (20 µg/ml) and concurrently with 5-FU (1 µg/ml) at 37°C for 24 h. For sequential treatment, cells were seeded in 6 dishes (1×105 in 3.5 cm-diameter dishes) and treated with LA (50 µg/ml) or EA (20 µg/ml) at 37°C for 24 h prior to 5-FU treatment (1 µg/ml at 37°C for 24 h).

The cell viabilities were assessed using a 3-(4, 5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfo-phenyl)-2H-tetrazolium (MTS) Assay kit (Promega Corporation, Madison, WI, USA). Following treatment with LA or EA and/or 5-FU, MTS solution was added to each well for 2 h at 37°C in 5% CO2. The absorbance at 490 nm was recorded using a microplate reader.

Animals

Male BALB/c mice (4-weeks-old) were purchased from Japan SLC, Inc. (Shizuoka, Japan). The animals were maintained in the pathogen-free animal facility with a 12/12 h light/dark cycle in a temperature (22°C) and humidity-controlled environment, according to institutional guidelines approved by the Committee for Animal Experimentation of Nara Medical University (Kashihara, Japan; approval no. 9559), in accordance with the current regulations and standards of the Japanese Ministry of Health, Labor and Welfare.

Animal models

To establish a subcutaneous tumor model, CT26 cancer cells (1×107) were inoculated into the scapular subcutaneous tissue of BALB/c mice. Mice were euthanized and observed using immunohistochemistry 4 weeks following inoculation. 5-FU (10 mg/kg; Wako Pure Chemical Industries, Ltd.) was injected intraperitoneally twice a week for 2 weeks following inoculation. LA (Sigma-Aldrich; Merck KGaA) or EA (Wako Pure Chemical Industries, Ltd.) were administrated at 1.0 and 0.1% w/w, respectively, through the daily supplementation of a standard CE-2 diet (CLEA Japan, Inc., Tokyo, Japan). The mean intake of LA and EA was 4.2±0.78 and 0.31±0.07 mg/day, respectively. A total of five mice were used for per treatment group. The mean weight of the mice was 21.5±1.4 g at the start of treatment.

Semi-quantitative reverse transcription-quantification polymerase chain reaction (RT-qPCR)

Total RNA (1 µg) was extracted using an RNeasy extraction kit (Qiagen, Inc., Valencia, CA, USA). RNA was extracted from 1×107 CT26 cells and eluted in 50 µl RNase-free water. cDNA was synthesized using ReverTra Ace RT-qPCR kit (Toyobo, Tokyo, Japan). Quantification of PCR products was performed with QuantiTect Primer Assays using QuantiFast, QuantiTect, Rotor-Gene, and FastLane kit for SYBR-Green-based detection (Qiagen, Inc.), according to the manufacturer's protocol. mRNA quantification was performed according to the 2−ΔΔcq method (19). The number of replicates was 30 cycles. The primer sets used for amplification were as follows: Mouse CD133 (prominin 1; accession no., BC028286.1) forward, 5′-GAAAAGTTGCTCTGCGAACC-3′ and reverse, 5′-TCTCAAGCTGAAAAGCAGCA-3′; mouse nucleostemin (NS; accession no. AY181025.1) forward, 5′-CAGGATGCTGACGATCAAGA-3′ and reverse, 5′-TTGATTGCTCAGGTGACAGC-3′; and mouse β-actin (accession no. NM_007393.4) forward, 5′-AGCCATGTACGTAGCCATCC-3′ and reverse 5′-CTCTCAGCTGTGGTGGTGAA-3′. The primers were synthesized by Sigma Genosys (Sigma-Aldrich; Merck KGaA). The PCR conditions were set according to the manufacturer's protocol. The number of replicates was 30 cycles. PCR products were electrophoresed on a 2% agarose gel and visualized using ethidium bromide.

Immunohistochemistry

Consecutive 4 µm tissue sections were immunohistochemically stained using the immunoperoxidase technique, as described previously (8). Mouse CD133 antibody (AG13328; ProteinTech Group, Inc., Chicago, IL, USA) and mouse nucleostemin antibody (sc-166430; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) were used at a concentration of 0.5 µg/ml at 37°C for 2 h. Color development was performed using 3–3′-diaminobenzidine (Dako; Agilent Technologies, Inc.), and the specimens were counterstained with Mayer's hematoxylin (Sigma-Aldrich; Merck KGaA) to visualize the nuclei. Following immunostaining, all slides were assessed to measure the number of positively stained nuclei using an all-in-one microscope (BZ-X700; Keyence, Osaka, Japan).

Aldehyde dehydrogenase (ALDH) activity

ALDH activity was measured using the ALDEFLUOR kit (Veritas Technologies LLC, Tokyo, Japan) according to the manufacturer's protocol. The activity was normalized to the negative control, diethylaminobenzaldehyde.

Statistical analysis

Statistical significance was calculated using the two-tailed Fisher's exact test, χ2 test and unpaired Student's t-test, with the assumption of Gaussian distributions, by Kolmogorov and Smirnov tests Statistical analysis was performed using InStat software (GraphPad Software, Inc., La Jolla, CA, USA). The data are presented as the mean ± standard deviation. P<0.05 (two-sided) was considered to indicate a statistically significant difference.

Results

Effect of LA and EA on the 5-FU-induced reduction of cancer cell viability

Three cancer cell lines, CT26 (colon cancer), LL2 (lung cancer) and CMT93 (rectal cancer), were concurrently treated with either LA or EA and 5-FU, or with LA, EA or 5-FU alone. Treatment with LA alone decreased cell viability in all 3 cell lines (Fig. 1). LA also augmented the tumor cell inhibition observed following 5-FU treatment in all 3 cell lines. In contrast, EA alone did not significantly suppress tumor viability in any of the 3 cell lines (Fig. 1). In addition, concurrent treatment of EA with 5-FU did not recover or enhance 5-FU-induced growth inhibition in the three cell lines.

Sequential treatment with LA or EA followed by 5-FU was then examined (Fig. 1). Concurrent treatment of 5-FU and LA revealed a more pronounced decrease of cell viability in the 3 cell lines. In contrast, temporal treatment of LA to 5-FU demonstrated no additional decrease of viability compared with that of 5-FU treatment alone. (Fig. 1). In addition, EA pretreatment reversed the effect of 5-FU, increasing the viability of LL2 and CMT93 cells (Fig. 1).

Effect of LA or EA with 5-FU on the stemness of cancer cells

Previous studies have demonstrated that the resistance of cancer cells to anticancer drugs is associated with CSCs (20,21). Therefore, alterations of stemness in LA- or EA-treated cancer cells were examined (Fig. 2). Expression of CD133 and NS was examined in cells treated with 5-FU alone, or concurrently with LA or EA (Fig. 2A-D). The expression of CD133 and NS was increased in cells treated with LA or EA when compared with untreated cells. Although expression of NS appears to be higher in cells treated with 5-FU and LA concurrently compared with cells treated with LA alone in CT26 and CMT93 cells, the expression of CD133 and NS was lower in cells treated concurrently with 5-FU and either LA or EA compared with cells treated with LA or EA alone. However expression was increased in cells treated concurrently with 5-FU and either LA or EA compared with cells treated with 5-FU alone. Activity of ALDH, a stem cell-associated enzyme, was increased in cells treated with LA or EA compared with in untreated cells (Fig. 2E). ALDH activity was higher in cells treated concurrently with 5-FU and either LA or EA compared with cells treated with 5-FU alone (Fig. 2E).

Effect of LA or EA with 5-FU on tumor growth

Finally, the effect of oral intake of LA or EA on the antitumor effects of 5-FU was examined in BALB/c mice inoculated with CT26 cells (Fig. 3). Treatment with 5-FU inhibited the growth of CT26-derived tumors, and this was abrogated by concurrent treatment with either LA or EA (Fig. 3A and B). The expression of NS in CT26-derived tumors was examined by immunohistochemistry (Fig. 3C and D). NS-positive CSCs were more abundant in tumors treated with 5-FU and either LA or EA compared with tumors treated with 5-FU alone.

Discussion

Fatty acids are dietary components, which are known to be involved in colon carcinogenesis (6,7). LA and EA are known to be risk factors for CRCs (14,2224), enhancing inflammation and suppressing mucosal immunity (2528). LA-derived prostaglandin E2, produced by cyclooxygenase-2, induces chronic persistent inflammation, which produces reactive oxygen species (25,26). Likewise, EA induces secretion of TNF-α, activation of nuclear factor-κB, and inhibition of cluster of differentiation 8+ T-lymphocytes (27,28).

LA and EA affect CSCs. LA induces senescence in cultured CRC cells and dormancy in CRC cells inoculated into mice (11). Conversely, EA increases the proliferation of cells in spheres via the activation of epidermal growth factor receptor and the Wnt signaling pathway, and the upregulation of stem cell markers (16,17). EA also induces epithelial-mesenchymal transition and enhances tumor growth and metastasis (16,17).

Previous studies have revealed that cancer stemness is responsible for resistance to anticancer chemotherapy (20,21). CSCs possess refractoriness to chemotherapy through low proliferative activity, advanced DNA repair and the expulsion of chemotherapeutic agents from the cytosol (29). Therefore, factors affecting the stemness of cancer cells may also affect the efficacy of chemotherapy.

Sequential treatment with LA or EA followed by 5-FU abrogated growth inhibition by 5-FU. Pretreatment with LA or EA may select and condense CSCs or increase the stemness of these cells. ALDH activity, a marker for stem cell activity (30), was enhanced by LA and EA. Upregulation of CD133 and NS by LA or EA varied by cell line. CD133 expression is associated with stem cells (31,32), whereas NS expression is associated with stem cells and progenitor cells (33). Therefore, LA and EA may affect populations of stem cells and progenitors in a cell or tissue-dependent manner (34).

The anti-chemotherapeutic effect of ingested LA or EA was confirmed using a mouse subcutaneous tumor model. Oral intake of LA or EA abrogated the antitumor effects of 5-FU. This result may be an example of the anti-chemotherapeutic properties of dietary components. The anti-chemotherapeutic effects of LA or EA were observed not only in CT26 and CMT93 CRC cells, but also in LL2 lung cancer cells. LA and EA are not thought to be risk factors for lung cancer; however, the present data indicated that the anti-chemotherapeutic effects of LA or EA may affect the efficacy of treatment of various malignancies. The effect of dietary components on chemotherapeutic effects, or on cancer stemness, should be the focus of wide ranging studies concerning human cancers.

Acknowledgements

The authors would like to thank Ms. Tomomi Masutani (Nara Medical University) for expert assistance with the preparation of this manuscript. The present study was supported by the Ministry of Education, Culture, Sports, Science and Technology KAKENHI (grant nos. 13200228, 14478268, 13394212, 13209774 and 16675788).

References

1 

Pietrzyk L, Torres A, Maciejewski R and Torres K: Obesity and obese-related chronic low-grade inflammation in promotion of colorectal cancer development. Asian Pac J Cancer Prev. 16:4161–4168. 2015. View Article : Google Scholar : PubMed/NCBI

2 

Uzunlulu M, Caklili O Telci and Oguz A: Association between metabolic syndrome and cancer. Ann Nutr Metab. 68:173–179. 2016. View Article : Google Scholar : PubMed/NCBI

3 

Cappellani A, Zanghi A, Di Vita M, Cavallaro A, Piccolo G, Veroux P, Lo Menzo E, Cavallaro V, De Paoli P, Veroux M and Berretta M: Strong correlation between diet and development of colorectal cancer. Front Biosci (Landmark Ed). 18:190–198. 2013. View Article : Google Scholar : PubMed/NCBI

4 

Rose DP: Dietary fatty acids and cancer. Am J Clin Nutr. 66 4 Suppl:998S–1003S. 1997.PubMed/NCBI

5 

Zhang W, Thornton WH and MacDonald RS: Insulin-like growth factor-I and II receptor expression in rat colon mucosa are affected by dietary lipid intake. J Nutr. 128:158–165. 1998.PubMed/NCBI

6 

Singh J, Hamid R and Reddy BS: Dietary fish oil inhibits the expression of farnesyl protein transferase and colon tumor development in rodents. Carcinogenesis. 19:985–989. 1998. View Article : Google Scholar : PubMed/NCBI

7 

Singh J, Hamid R and Reddy BS: Dietary fat and colon cancer: Modulation of cyclooxygenase-2 by types and amount of dietary fat during the postinitiation stage of colon carcinogenesis. Cancer Res. 57:3465–3470. 1997.PubMed/NCBI

8 

Ohmori H, Luo Y, Fujii K, Sasahira T, Shimomoto T, Denda A and Kuniyasu H: Dietary linoleic acid and glucose enhances azoxymethane-induced colon cancer and metastases via the expression of high-mobility group box 1. Pathobiology. 77:210–217. 2010. View Article : Google Scholar : PubMed/NCBI

9 

Zhou S, Wang G, Chen B and Wang P: Effect of dietary fatty acids on tumorigenesis of colon cancer induced by methylnitrosourea in rats. J Environ Pathol Toxicol Oncol. 19:81–86. 2000.PubMed/NCBI

10 

Rao CV, Hirose Y, Indranie C and Reddy BS: Modulation of experimental colon tumorigenesis by types and amounts of dietary fatty acids. Cancer Res. 61:1927–1933. 2001.PubMed/NCBI

11 

Ohmori H, Sasahira T, Fujii K, Yi L, Shimomoto T and Kuniyasu H: Linoleic acid-induced growth suppression induces quiescent cancer cell nests in nude mice. Pathobiology. 75:226–232. 2008. View Article : Google Scholar : PubMed/NCBI

12 

Luo Y, Chihara Y, Fujimoto K, Sasahira T, Kuwada M, Fujiwara R, Fujii K, Ohmori H and Kuniyasu H: High mobility group box 1 released from necrotic cells enhances regrowth and metastasis of cancer cells that have survived chemotherapy. Eur J Cancer. 49:741–751. 2013. View Article : Google Scholar : PubMed/NCBI

13 

King IB, Kristal AR, Schaffer S, Thornquist M and Goodman GE: Serum trans-fatty acids are associated with risk of prostate cancer in beta-Carotene and Retinol Efficacy Trial. Cancer Epidemiol Biomarkers Prev. 14:988–992. 2005. View Article : Google Scholar : PubMed/NCBI

14 

Vinikoor LC, Schroeder JC, Millikan RC, Satia JA, Martin CF, Ibrahim J, Galanko JA and Sandler RS: Consumption of trans-fatty acid and its association with colorectal adenomas. Am J Epidemiol. 168:289–297. 2008. View Article : Google Scholar : PubMed/NCBI

15 

Kuniyasu H, Ohmori H, Fujii K, et al: Significance of trans fatty acids in colorectal cancerCurrent Research in Cancer. 9. Sunitha M: Research Media; Kerala, India: pp. 21–29. 2015

16 

Ohmori H, Fujii K, Kadochi Y, Mori S, Nishiguchi Y, Fujiwara R, Kishi S, Sasaki T and Kuniyasu H: Elaidic acid, a trans fatty acid, enhances the metastasis of colorectal cancer cells. Pathobiology. 84:144–151. 2017. View Article : Google Scholar : PubMed/NCBI

17 

Fujii K, Luo Y, Fujiwara-Tani R, Kishi S, He S, Yang S, Sasaki T, Ohmori H and Kuniyasu H: Pro-metastatic intracellular signaling of elaidic trans fatty acid. Int J Oncol. 50:85–92. 2017.PubMed/NCBI

18 

U.S. Food and Drug Administration, . Final Determination Regarding Partially Hydrogenated Oils. U.S. Food and Drug Administration; Silver Spring, MD: 2015

19 

Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI

20 

Leirós GJ and Balañá ME: Metastatic cancer stem cells: New molecular targets for cancer therapy. Curr Pharm Biotechnol. 12:1909–1922. 2011. View Article : Google Scholar : PubMed/NCBI

21 

Gaur P, Sceusi EL, Samuel S, Xia L, Fan F, Zhou Y, Lu J, Tozzi F, Lopez-Berestein G, Vivas-Mejia P, et al: Identification of cancer stem cells in human gastrointestinal carcinoid and neuroendocrine tumors. Gastroenterology. 141:1728–1737. 2011. View Article : Google Scholar : PubMed/NCBI

22 

Bultman SJ: Interplay between diet, gut microbiota, epigenetic events and colorectal cancer. Mol Nutr Food Res. 61:2017. View Article : Google Scholar : PubMed/NCBI

23 

Rao CV and Reddy BS: Modulating effect of amount and types of dietary fat on ornithine decarboxylase, tyrosine protein kinase and prostaglandins production during colon carcinogenesis in male F344 rats. Carcinogenesis. 14:1327–1333. 1993. View Article : Google Scholar : PubMed/NCBI

24 

Slattery ML, Benson J, Ma KN, Schaffer D and Potter JD: Trans-fatty acids and colon cancer. Nutr Cancer. 39:170–175. 2001. View Article : Google Scholar : PubMed/NCBI

25 

Gupta RA and Dubois RN: Colorectal cancer prevention and treatment by inhibition of cyclooxygenase-2. Nat Rev Cancer. 1:11–21. 2001. View Article : Google Scholar : PubMed/NCBI

26 

Moonen HJ, Dommels YE, van Zwam M, van Herwijnen MH, Kleinjans JC, Alink GM and de Kok TM: Effects of polyunsaturated fatty acids on prostaglandin synthesis and cyclooxygenase-mediated DNA adduct formation by heterocyclic aromatic amines in human adenocarcinoma colon cells. Mol Carcinog. 40:180–188. 2004. View Article : Google Scholar : PubMed/NCBI

27 

Rao YP, Kumar PP and Lokesh BR: Molecular mechanisms for the modulation of selected inflammatory markers by dietary rice bran oil in rats fed partially hydrogenated vegetable fat. Lipids. 51:451–467. 2016. View Article : Google Scholar : PubMed/NCBI

28 

Dlouhý P, Kucera P, Kraml P, Pompachová A, Potocková J, Smejkalová V, Mokrejs P, Jacek M and Andel M: Short-term dietary intake of C18:1 trans fatty acids decreases the function of cellular immunity in healthy young men. Ann Nutr Metab. 53:129–136. 2008. View Article : Google Scholar : PubMed/NCBI

29 

McCubrey JA, Abrams SL, Fitzgerald TL, Cocco L, Martelli AM, Montalto G, Cervello M, Scalisi A, Candido S, Libra M and Steelman LS: Roles of signaling pathways in drug resistance, cancer initiating cells and cancer progression and metastasis. Adv Biol Regul. 57:75–101. 2015. View Article : Google Scholar : PubMed/NCBI

30 

Meng E, Mitra A, Tripathi K, Finan MA, Scalici J, McClellan S, da Silva L Madeira, Reed E, Shevde LA, Palle K and Rocconi RP: ALDH1A1 maintains ovarian cancer stem cell-like properties by altered regulation of cell cycle checkpoint and DNA repair network signaling. PLoS One. 9:e1071422014. View Article : Google Scholar : PubMed/NCBI

31 

Ren F, Sheng WQ and Du X: CD133: A cancer stem cells marker, is used in colorectal cancers. World J Gastroenterol. 19:2603–2611. 2013. View Article : Google Scholar : PubMed/NCBI

32 

Sun Y, Kong W, Falk A, Hu J, Zhou L, Pollard S and Smith A: CD133 (Prominin) negative human neural stem cells are clonogenic and tripotent. PLoS One. 4:e54982009. View Article : Google Scholar : PubMed/NCBI

33 

Liu SJ, Cai ZW, Liu YJ, Dong MY, Sun LQ, Hu GF, Wei YY and Lao WD: Role of nucleostemin in growth regulation of gastric cancer, liver cancer and other malignancies. World J Gastroenterol. 10:1246–1249. 2004.PubMed/NCBI

34 

Haraguchi N, Inoue H, Tanaka F, Mimori K, Utsunomiya T, Sasaki A and Mori M: Cancer stem cells in human gastrointestinal cancers. Hum Cell. 19:24–29. 2006. View Article : Google Scholar : PubMed/NCBI

Related Articles

Journal Cover

July 2017
Volume 14 Issue 1

Print ISSN: 1792-1074
Online ISSN:1792-1082

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
APA
Tanabe, E., Kitayoshi, M., Fujii, K., Ohmori, H., Luo, Y., Kadochi, Y. ... Kuniyasu, H. (2017). Fatty acids inhibit anticancer effects of 5-fluorouracil in mouse cancer cell lines. Oncology Letters, 14, 681-686. https://doi.org/10.3892/ol.2017.6190
MLA
Tanabe, E., Kitayoshi, M., Fujii, K., Ohmori, H., Luo, Y., Kadochi, Y., Mori, S., Fujiwara, R., Nishiguchi, Y., Sasaki, T., Kuniyasu, H."Fatty acids inhibit anticancer effects of 5-fluorouracil in mouse cancer cell lines". Oncology Letters 14.1 (2017): 681-686.
Chicago
Tanabe, E., Kitayoshi, M., Fujii, K., Ohmori, H., Luo, Y., Kadochi, Y., Mori, S., Fujiwara, R., Nishiguchi, Y., Sasaki, T., Kuniyasu, H."Fatty acids inhibit anticancer effects of 5-fluorouracil in mouse cancer cell lines". Oncology Letters 14, no. 1 (2017): 681-686. https://doi.org/10.3892/ol.2017.6190