Loss of Runt-related transcription factor 3 induces resistance to 5-fluorouracil and cisplatin in hepatocellular carcinoma

Kataoka,Junro; Shiraha,Hidenori; Horiguchi,Shigeru; Sawahara,Hiroaki; Uchida,Daisuke; Nagahara,Teruya; Iwamuro,Masaya; Morimoto,Hiroki; Takeuchi,Yasuto; Kuwaki,Kenji; Onishi,Hideki; Nakamura,Shinichiro; Takaki,Akinobu; Nouso,Kazuhiro; Yagi,Takahito; Yamamoto,Kazuhide; Okada,Hiroyuki

doi:10.3892/or.2016.4681

Loss of Runt-related transcription factor 3 induces resistance to 5-fluorouracil and cisplatin in hepatocellular carcinoma

Authors:
- Junro Kataoka
- Hidenori Shiraha
- Shigeru Horiguchi
- Hiroaki Sawahara
- Daisuke Uchida
- Teruya Nagahara
- Masaya Iwamuro
- Hiroki Morimoto
- Yasuto Takeuchi
- Kenji Kuwaki
- Hideki Onishi
- Shinichiro Nakamura
- Akinobu Takaki
- Kazuhiro Nouso
- Takahito Yagi
- Kazuhide Yamamoto
- Hiroyuki Okada
View Affiliations

Affiliations: Department of Gastroenterology and Hepatology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700-8558, Japan, Department of Gastroenterological Surgery, Transplant and Surgical Oncology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700-8558, Japan, Department of Gastroenterology, Okayama Saiseikai General Hospital, Okayama 700-8511, Japan
Published online on: March 16, 2016 https://doi.org/10.3892/or.2016.4681
Pages: 2576-2582
Copyright: © Kataoka 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: )

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

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Runt-related transcription factor 3 (RUNX3) is known to function as a tumor suppressor in gastric cancer and other types of cancers, including hepatocellular carcinoma (HCC). However, its role has not been fully elucidated. In the present study, we aimed to evaluate the role of RUNX3 in HCC. We used the human HCC cell lines Hep3B, Huh7 and HLF; RUNX3 cDNA was introduced into Hep3B and Huh7 cells, which were negative for endogenous RUNX3 expression, and RUNX3 siRNA was transfected into HLF cells, which were positive for endogenous RUNX3. We analyzed the expression of RUNX3 and multidrug resistance-associated protein (MRP) by immunoblotting. MTT assays were used to determine the effects of RUNX3 expression on 5-fluorouracil (5-FU) and cisplatin (CDDP) sensitivity. Finally, 23 HCC specimens resected from patients with HCC at Okayama University Hospital were analyzed, and correlations among immunohistochemical expression of RUNX3 protein and MRP protein were evaluated in these specimens. Exogenous RUNX3 expression reduced the expression of MRP1, MRP2, MRP3 and MRP5 in the RUNX3-negative cells, whereas knockdown of RUNX3 in the HLF cells stimulated the expression of these MRPs. An inverse correlation between RUNX3 and MRP expression was observed in the HCC tissues. Importantly, loss of RUNX3 expression contributed to 5-FU and CDDP resistance by inducing MRP expression. These data have important implications in the study of chemotherapy resistance in HCC.

View Figures

View References

1	El-Serag HB and Rudolph KL: Hepatocellular carcinoma: Epidemiology and molecular carcinogenesis. Gastroenterology. 132:2557–2576. 2007. View Article : Google Scholar : PubMed/NCBI
2	El-Serag HB: Hepatocellular carcinoma. N Engl J Med. 365:1118–1127. 2011. View Article : Google Scholar : PubMed/NCBI
3	Arumugam T, Ramachandran V, Fournier KF, Wang H, Marquis L, Abbruzzese JL, Gallick GE, Logsdon CD, McConkey DJ and Choi W: Epithelial to mesenchymal transition contributes to drug resistance in pancreatic cancer. Cancer Res. 69:5820–5828. 2009. View Article : Google Scholar : PubMed/NCBI
4	Cheng L, Luo S, Jin C, Ma H, Zhou H and Jia L: FUT family mediates the multidrug resistance of human hepatocellular carcinoma via the PI3K/Akt signaling pathway. Cell Death Dis. 4:e9232013. View Article : Google Scholar : PubMed/NCBI
5	Marin JJ, Monte MJ, Blazquez AG, Macias RI, Serrano MA and Briz O: The role of reduced intracellular concentrations of active drugs in the lack of response to anticancer chemotherapy. Acta Pharmacol Sin. 35:1–10. 2014. View Article : Google Scholar :
6	Dean M, Hamon Y and Chimini G: The human ATP-binding cassette (ABC) transporter superfamily. J Lipid Res. 42:1007–1017. 2001.PubMed/NCBI
7	Hagmann W, Jesnowski R, Faissner R, Guo C and Löhr JM: ATP-binding cassette C transporters in human pancreatic carcinoma cell lines. Upregulation in 5-fluorouracil-resistant cells. Pancreatology. 9:136–144. 2009. View Article : Google Scholar
8	Nambaru PK, Hübner T, Köck K, Mews S, Grube M, Payen L, Guitton J, Sendler M, Jedlitschky G, Rimmbach C, et al: Drug efflux transporter multidrug resistance-associated protein 5 affects sensitivity of pancreatic cancer cell lines to the nucleoside anticancer drug 5-fluorouracil. Drug Metab Dispos. 39:132–139. 2011. View Article : Google Scholar
9	Hagmann W, Faissner R, Schnölzer M, Löhr M and Jesnowski R: Membrane drug transporters and chemoresistance in human pancreatic carcinoma. Cancers. 3:106–125. 2010. View Article : Google Scholar : PubMed/NCBI
10	Hlavata I, Mohelnikova-Duchonova B, Vaclavikova R, Liska V, Pitule P, Novak P, Bruha J, Vycital O, Holubec L, Treska V, et al: The role of ABC transporters in progression and clinical outcome of colorectal cancer. Mutagenesis. 27:187–196. 2012. View Article : Google Scholar : PubMed/NCBI
11	Long J, Zhang Y, Yu X, Yang J, LeBrun DG, Chen C, Yao Q and Li M: Overcoming drug resistance in pancreatic cancer. Expert Opin Ther Targets. 15:817–828. 2011. View Article : Google Scholar : PubMed/NCBI
12	Wang WB, Yang Y, Zhao YP, Zhang TP, Liao Q and Shu H: Recent studies of 5-fluorouracil resistance in pancreatic cancer. World J Gastroenterol. 20:15682–15690. 2014. View Article : Google Scholar : PubMed/NCBI
13	Park JY, Ahn SH, Yoon YJ, Kim JK, Lee HW, Lee Y, Chon CY, Moon YM and Han KH: Repetitive short-course hepatic arterial infusion chemotherapy with high-dose 5-fluorouracil and cisplatin in patients with advanced hepatocellular carcinoma. Cancer. 110:129–137. 2007. View Article : Google Scholar : PubMed/NCBI
14	Ma MC, Chen YY, Li SH, Cheng YF, Wang CC, Chiu TJ, Pei SN, Liu CT, Huang TL, Huang CH, et al: Intra-arterial chemotherapy with doxorubicin and cisplatin is effective for advanced hepatocellular cell carcinoma. Scientific World Journal. 2014:1601382014. View Article : Google Scholar : PubMed/NCBI
15	Johnson PJ: Are there indications for chemotherapy in hepatocellular carcinoma? Surg Oncol Clin N Am. 12:127–134. 2003. View Article : Google Scholar : PubMed/NCBI
16	Nishimura M: A successful treatment by hepatic arterial infusion therapy for advanced, unresectable biliary tract cancer. World J Hepatol. 2:192–197. 2010.PubMed/NCBI
17	Shen DW, Pouliot LM, Hall MD and Gottesman MM: Cisplatin resistance: A cellular self-defense mechanism resulting from multiple epigenetic and genetic changes. Pharmacol Rev. 64:706–721. 2012. View Article : Google Scholar : PubMed/NCBI
18	Kowalski P, Surowiak P and Lage H: Reversal of different drug-resistant phenotypes by an autocatalytic multitarget multiribozyme directed against the transcripts of the ABC transporters MDR1/P-gp, MRP2, and BCRP. Mol Ther. 11:508–522. 2005. View Article : Google Scholar : PubMed/NCBI
19	Kuwano M, Toh S, Uchiumi T, Takano H, Kohno K and Wada M: Multidrug resistance-associated protein subfamily transporters and drug resistance. Anticancer Drug Des. 14:123–131. 1999.PubMed/NCBI
20	Materna V, Holm PS, Dietel M and Lage H: Kinetic characterization of ribozymes directed against the cisplatin resistance-associated ABC transporter cMOAT/MRP2/ABCC2. Cancer Gene Ther. 8:176–184. 2001. View Article : Google Scholar : PubMed/NCBI
21	Li QL, Ito K, Sakakura C, Fukamachi H, Inoue K, Chi XZ, Lee KY, Nomura S, Lee CW, Han SB, et al: Causal relationship between the loss of RUNX3 expression and gastric cancer. Cell. 109:113–124. 2002. View Article : Google Scholar : PubMed/NCBI
22	Subramaniam MM, Chan JY, Yeoh KG, Quek T, Ito K and Salto-Tellez M: Molecular pathology of RUNX3 in human carcinogenesis. Biochim Biophys Acta. 1796:315–331. 2009.PubMed/NCBI
23	Nakanishi Y, Shiraha H, Nishina S, Tanaka S, Matsubara M, Horiguchi S, Iwamuro M, Takaoka N, Uemura M, Kuwaki K, et al: Loss of runt-related transcription factor 3 expression leads hepatocellular carcinoma cells to escape apoptosis. BMC Cancer. 11:32011. View Article : Google Scholar : PubMed/NCBI
24	Nishina S, Shiraha H, Nakanishi Y, Tanaka S, Matsubara M, Takaoka N, Uemura M, Horiguchi S, Kataoka J, Iwamuro M, et al: Restored expression of the tumor suppressor gene RUNX3 reduces cancer stem cells in hepatocellular carcinoma by suppressing Jagged1-Notch signaling. Oncol Rep. 26:523–531. 2011.PubMed/NCBI
25	Tanaka S, Shiraha H, Nakanishi Y, Nishina S, Matsubara M, Horiguchi S, Takaoka N, Iwamuro M, Kataoka J, Kuwaki K, et al: Runt-related transcription factor 3 reverses epithelial-mesenchymal transition in hepatocellular carcinoma. Int J Cancer. 131:2537–2546. 2012. View Article : Google Scholar : PubMed/NCBI
26	Horiguchi S, Shiraha H, Nagahara T, Kataoka J, Iwamuro M, Matsubara M, Nishina S, Kato H, Takaki A, Nouso K, et al: Loss of runt-related transcription factor 3 induces gemcitabine resistance in pancreatic cancer. Mol Oncol. 7:840–849. 2013. View Article : Google Scholar : PubMed/NCBI
27	Mori T, Nomoto S, Koshikawa K, Fujii T, Sakai M, Nishikawa Y, Inoue S, Takeda S, Kaneko T and Nakao A: Decreased expression and frequent allelic inactivation of the RUNX3 gene at 1p36 in human hepatocellular carcinoma. Liver Int. 25:380–388. 2005. View Article : Google Scholar : PubMed/NCBI
28	Xiao WH and Liu WW: Hemizygous deletion and hypermethylation of RUNX3 gene in hepatocellular carcinoma. World J Gastroenterol. 10:376–380. 2004.PubMed/NCBI
29	Li X, Zhang Y, Zhang Y, Qiao T, Wu K, Ding J, Liu J and Fan D: RUNX3 inhibits growth of HCC cells and HCC xenografts in mice in combination with adriamycin. Cancer Biol Ther. 7:669–676. 2008. View Article : Google Scholar : PubMed/NCBI
30	Guo C, Ding J, Yao L, Sun L, Lin T, Song Y, Sun L and Fan D: Tumor suppressor gene Runx3 sensitizes gastric cancer cells to chemotherapeutic drugs by downregulating Bcl-2, MDR-1 and MRP-1. Int J Cancer. 116:155–160. 2005. View Article : Google Scholar : PubMed/NCBI
31	Lu F, Hou YQ, Song Y and Yuan ZJ: TFPI-2 downregulates multidrug resistance protein in 5-FU-resistant human hepatocellular carcinoma BEL-7402/5-FU cells. Anat Rec. 296:56–63. 2013. View Article : Google Scholar
32	Gu W, Fang FF, Li B, Cheng BB and Ling CQ: Characterization and resistance mechanisms of a 5-fluorouracil-resistant hepatocellular carcinoma cell line. Asian Pac J Cancer Prev. 13:4807–4814. 2012. View Article : Google Scholar
33	Zisowsky J, Koegel S, Leyers S, Devarakonda K, Kassack MU, Osmak M and Jaehde U: Relevance of drug uptake and efflux for cisplatin sensitivity of tumor cells. Biochem Pharmacol. 73:298–307. 2007. View Article : Google Scholar
34	Siddik ZH: Cisplatin: Mode of cytotoxic action and molecular basis of resistance. Oncogene. 22:7265–7279. 2003. View Article : Google Scholar : PubMed/NCBI
35	Samimi G, Katano K, Holzer AK, Safaei R and Howell SB: Modulation of the cellular pharmacology of cisplatin and its analogs by the copper exporters ATP7A and ATP7B. Mol Pharmacol. 66:25–32. 2004. View Article : Google Scholar : PubMed/NCBI