PMID- 28587170 OWN - NLM STAT- MEDLINE DCOM- 20180326 LR - 20181202 IS - 1422-0067 (Electronic) IS - 1422-0067 (Linking) VI - 18 IP - 6 DP - 2017 Jun 1 TI - The Combination of Arginine Deprivation and 5-Fluorouracil Improves Therapeutic Efficacy in Argininosuccinate Synthetase Negative Hepatocellular Carcinoma. LID - 10.3390/ijms18061175 [doi] LID - 1175 AB - Argininosuccinate synthetase (ASS), a key enzyme to synthesize arginine is down regulated in many tumors including hepatocellular carcinoma (HCC). Similar to previous reports, we have found the decrease in ASS expression in poorly differentiated HCC. These ASS(-) tumors are auxotrophic for arginine. Pegylated arginine deiminase (ADI-PEG20), which degrades arginine, has shown activity in these tumors, but the antitumor effect is not robust and hence combination treatment is needed. Herein, we have elucidated the effectiveness of ADI-PEG20 combined with 5-Fluorouracil (5-FU) in ASS(-)HCC by targeting urea cycle and pyrimidine metabolism using four HCC cell lines as model. SNU398 and SNU387 express very low levels of ASS or ASS(-) while Huh-1, and HepG2 express high ASS similar to normal cells. Our results showed that the augmented cytotoxic effect of combination treatment only occurs in SNU398 and SNU387, and not in HepG2 and Huh-1 (ASS(+)) cells, and is partly due to reduced anti-apoptotic proteins X-linked inhibitor of apoptosis protein (XIAP), myeloid leukemia cell differentiation protein (Mcl-1) and B-cell lymphoma-2 (Bcl-2). Importantly, lack of ASS also influences essential enzymes in pyrimidine synthesis (carbamoyl-phosphate synthetase2, aspartate transcarbamylase and dihydrooratase (CAD) and thymidylate synthase (TS)) and malate dehydrogenase-1 (MDH-1) in TCA cycle. ADI-PEG20 treatment decreased these enzymes and made them more vulnerable to 5-FU. Transfection of ASS restored these enzymes and abolished the sensitivity to ADI-PEG20 and combination treatment. Overall, our data suggest that ASS influences multiple enzymes involved in 5-FU sensitivity. Combining ADI-PEG20 and 5-FU may be effective to treat ASS(-)hepatoma and warrants further clinical investigation. FAU - Thongkum, Angkana AU - Thongkum A AD - Laboratory of Environmental Toxicology, Chulabhorn Research Institute, Laksi, Bangkok 10210, Thailand. d10120101@cgi.ac.th. AD - Chulabhorn Graduate Institute, Laksi, Bangkok 10210, Thailand. d10120101@cgi.ac.th. FAU - Wu, Chunjing AU - Wu C AD - Division of Hematology/Oncology, Miami Veterans Affairs Healthcare System, Miami, FL 33125, USA. chunjingwu@hotmail.com. FAU - Li, Ying-Ying AU - Li YY AD - Division of Hematology/Oncology, Miami Veterans Affairs Healthcare System, Miami, FL 33125, USA. YLi4@med.miami.edu. AD - Sylvester Comprehensive Cancer Center, University of Miami Miller School of Medicine, Miami, FL 33136, USA. YLi4@med.miami.edu. FAU - Wangpaichitr, Medhi AU - Wangpaichitr M AD - Division of Hematology/Oncology, Miami Veterans Affairs Healthcare System, Miami, FL 33125, USA. Mwangpaichitre@med.miami.edu. AD - Department of Surgery, University of Miami Miller School of Medicine, Miami, FL 33125, USA. Mwangpaichitre@med.miami.edu. FAU - Navasumrit, Panida AU - Navasumrit P AD - Laboratory of Environmental Toxicology, Chulabhorn Research Institute, Laksi, Bangkok 10210, Thailand. Panida@cri.or.th. AD - Chulabhorn Graduate Institute, Laksi, Bangkok 10210, Thailand. Panida@cri.or.th. AD - Center of Excellence on Environmental Health, Toxicology (EHT), Ministry of Education, Bangkok 10300, Thailand. Panida@cri.or.th. FAU - Parnlob, Varabhorn AU - Parnlob V AD - Laboratory of Environmental Toxicology, Chulabhorn Research Institute, Laksi, Bangkok 10210, Thailand. varabhorn@cri.or.th. FAU - Sricharunrat, Thaniya AU - Sricharunrat T AD - Laboratory Unit of Pathology, Chulabhorn Hospital, Laksi, Bangkok 10210, Thailand. sri.thaniya@gmail.com. FAU - Bhudhisawasdi, Vajarabhongsa AU - Bhudhisawasdi V AD - Department of Surgery, Faculty of Medicine, Khonkaen University, Khonkaen 40000, Thailand. joevajara@gmail.com. AD - Laboratory of Chemical Carcinogenesis, Chulabhorn Research Institute, Laksi, Bangkok 10210, Thailand. joevajara@gmail.com. FAU - Ruchirawat, Mathuros AU - Ruchirawat M AD - Laboratory of Environmental Toxicology, Chulabhorn Research Institute, Laksi, Bangkok 10210, Thailand. mathuros@cri.or.th. AD - Center of Excellence on Environmental Health, Toxicology (EHT), Ministry of Education, Bangkok 10300, Thailand. mathuros@cri.or.th. FAU - Savaraj, Niramol AU - Savaraj N AD - Division of Hematology/Oncology, Miami Veterans Affairs Healthcare System, Miami, FL 33125, USA. nsavaraj@med.miami.edu. AD - Sylvester Comprehensive Cancer Center, University of Miami Miller School of Medicine, Miami, FL 33136, USA. nsavaraj@med.miami.edu. LA - eng GR - I01 BX003328/BX/BLRD VA/United States PT - Journal Article DEP - 20170601 PL - Switzerland TA - Int J Mol Sci JT - International journal of molecular sciences JID - 101092791 RN - 0 (Antineoplastic Agents) RN - 3WJQ0SDW1A (Polyethylene Glycols) RN - 94ZLA3W45F (Arginine) RN - EC 3.- (Hydrolases) RN - EC 3.5.3.6 (ADI PEG20) RN - EC 6.3.4.5 (Argininosuccinate Synthase) RN - U3P01618RT (Fluorouracil) SB - IM MH - Adult MH - Aged MH - Antineoplastic Agents/pharmacology/therapeutic use MH - Antineoplastic Combined Chemotherapy Protocols/therapeutic use MH - Apoptosis/drug effects/genetics MH - Arginine/*metabolism MH - Argininosuccinate Synthase/*deficiency/genetics/metabolism MH - Carcinoma, Hepatocellular/*drug therapy/*metabolism/pathology MH - Cell Line, Tumor MH - Cell Proliferation/drug effects MH - Female MH - Fluorouracil/pharmacology/*therapeutic use MH - Gene Expression MH - Gene Expression Regulation, Enzymologic MH - Gene Expression Regulation, Neoplastic MH - Humans MH - Hydrolases/pharmacology MH - Liver Neoplasms/*drug therapy/*metabolism/pathology MH - Male MH - Middle Aged MH - Models, Biological MH - Polyethylene Glycols/pharmacology MH - Treatment Outcome PMC - PMC5485998 OTO - NOTNLM OT - 5-FU OT - ADI-PEG20 OT - ASS re-expression OT - ASS(-)Hepatocellular carcinoma OT - pyrimidine metabolism OT - thymidylate synthase COIS- The authors declare no conflict of interest. EDAT- 2017/06/08 06:00 MHDA- 2018/03/27 06:00 PMCR- 2017/06/01 CRDT- 2017/06/08 06:00 PHST- 2017/03/30 00:00 [received] PHST- 2017/05/18 00:00 [revised] PHST- 2017/05/26 00:00 [accepted] PHST- 2017/06/08 06:00 [entrez] PHST- 2017/06/08 06:00 [pubmed] PHST- 2018/03/27 06:00 [medline] PHST- 2017/06/01 00:00 [pmc-release] AID - ijms18061175 [pii] AID - ijms-18-01175 [pii] AID - 10.3390/ijms18061175 [doi] PST - epublish SO - Int J Mol Sci. 2017 Jun 1;18(6):1175. doi: 10.3390/ijms18061175.