Dichloroacetate enhances the antitumor effect of pirarubicin via regulating the ROS-JNK signaling pathway in liver cancer cells

Abstract

Aim: Liver cancer is one of the most common malignancies and has a high recurrence rate. However, current treatment strategies do not achieve satisfactory outcomes in the clinic. To explore a new strategy to enhance the effectiveness of chemotherapy in liver cancer, we investigated whether dichloroacetate (DCA) could enhance the sensitivity of liver cancer cells to pirarubicin (THP).

Methods: Liver cancer cells were treated with DCA alone, THP alone, or DCA and THP combined. Cell viability was determined by the CCK-8 assay. Cell apoptosis was analyzed by flow cytometer. Reactive oxygen species (ROS) were detected using a CM-H2DCFDA fluorescence probe. Protein levels were identified by immunoblotting.

Results: The results revealed that DCA significantly enhanced the antitumor effect of THP in liver cancer cells. Changes in morphology and adherence ability were observed, as well as decreased cell viability. The results of flow cytometry showed that the combination of THP and DCA significantly increased apoptosis of liver cancer cells. Moreover, compared with THP alone, combination treatment with DCA significantly increased THP-triggered ROS generation in liver cancer cells. The antioxidant N-acetyl-L-cysteine reversed the synergistic effect of DCA and THP on ROS generation, cell viability and apoptosis. Furthermore, phosphorylation of c-Jun N-terminal kinase (JNK) was significantly increased in the DCA and THP combination group. The effects of DCA and THP on cell viability and apoptosis were inhibited by the JNK inhibitor SP600125.

Conclusion: The results obtained in the present study indicated that DCA enhanced the antitumor effect of THP in liver cancer cells via regulating the ROS-JNK signaling pathway.

Keywords

Dichloroacetate / c-Jun N-terminal kinase / liver cancer cells / pirarubicin / reactive oxygen species

References

[1] Khemlina G,Kurzrock R.The biology of Hepatocellular carcinoma: implications for genomic and immune therapies..Mol Cancer2017;16:149 PMCID:PMC5577674

[2] Lee DH.Primary malignant tumours in the non-cirrhotic liver..Eur J Radiol2017;95:349-61

[3] Tsurumi H,Goto N,Kasahara S.A phase II study of a THP-COP regimen for the treatment of elderly patients aged 70 years or older with diffuse large B-cell lymphoma..Hematol Oncol2007;25:107-14

[4] Daruwalla J,Nikfarjam M,Malcontenti-Wilson C.Evaluation of the effect of SMA-pirarubicin micelles on colorectal cancer liver metastases and of hyperbaric oxygen in CBA mice..J Drug Target2007;15:487-95

[5] Zou HY,Zhang Y,Nie JF.Studying the interaction of pirarubicin with DNA and determining pirarubicin in human urine samples: combining excitation-emission fluorescence matrices with second-order calibration methods..J Fluoresc2009;19:955-66

[6] Sieghart W,Pinter M,Vogel W.The ART of decision making: retreatment with transarterial chemoembolization in patients with hepatocellular carcinoma..Hepatology2013;57:2261-73

[7] Georgiades C,Harrison N,Liapi E.Lack of response after initial chemoembolization for hepatocellular carcinoma: does it predict failure of subsequent treatment?.Radiology2012;265:115-23 PMCID:PMC4137783

[8] Li B,Sun Q,Chen L.Reversal of the Warburg effect with DCA in PDGFtreated human PASMC is potentiated by pyruvate dehydrogenase kinase1 inhibition mediated through blocking Akt/GSK3beta signalling..Int J Mol Med2018;42:1391-400 PMCID:PMC6089770

[9] Peng F,Fan WJ,Li MM.Glycolysis gatekeeper PDK1 reprograms breast cancer stem cells under hypoxia..Oncogene2018;37:1062-74 PMCID:PMC5851116

[10] Kan PC,Chien CS,Fang HW.Coupling dichloroacetate treatment with curcumin significantly enhances anticancer potential..Anticancer Res2018;38:6253-61

[11] Shen YC,Hsu C,Chang CY.Activating oxidative phosphorylation by a pyruvate dehydrogenase kinase inhibitor overcomes sorafenib resistance of hepatocellular carcinoma..Br J Cancer2013;108:72-81 PMCID:PMC3553537

[12] Li P,Lin C,Zhu X.The synergistic effect of propofol and ulinastatin suppressed the viability of the human lung adenocarcinoma epithelial A549 cell line..Oncol Lett2018;16:5191-9 PMCID:PMC6144888

[13] Chou TC.Drug combination studies and their synergy quantification using the Chou-Talalay method..Cancer Res2010;70:440-6

[14] Ni Z,Dai X,Yang T.HCC cells with high levels of Bcl-2 are resistant to ABT-737 via activation of the ROS-JNK-autophagy pathway..Free Radic Biol Med2014;70:194-203

[15] Nowak R.Anthraquinone antitumour agents, doxorubicin, pirarubicin and benzoperimidine BP1, trigger caspase-3/caspase-8-dependent apoptosis of leukaemia sensitive HL60 and resistant HL60/VINC and HL60/DOX cells..Anticancer Drugs2012;23:380-92

[16] Daruwalla J,Greish K,Muralidharan V.In vitro and in vivo evaluation of tumor targeting styrene-maleic acid copolymer-pirarubicin micelles: Survival improvement and inhibition of liver metastases..Cancer Sci2010;101:1866-74

[17] Liang YH,Chen JY,Cheng AL.Modern prospection for hepatic arterial infusion chemotherapy in malignancies with liver metastases..Int J Hepatol2013;2013:141590 PMCID:PMC3652147

[18] Hirakawa M,Asayama Y,Ishigami K.Efficacy of preoperative transcatheter arterial chemoembolization combined with systemic chemotherapy for treatment of unresectable hepatoblastoma in children..Jpn J Radiol2014;32:529-36

[19] James MO JS,Smeltz MG,Stacpoole PW.Therapeutic applications of dichloroacetate and the role of glutathione transferase zeta-1..Pharmacol Ther2017;170:15 PMCID:PMC5274567

[20] Tataranni T.Dichloroacetate (DCA) and cancer: an overview towards clinical applications..Oxid Med Cell Longev2019;2019:8201079 PMCID:PMC6885244

[21] Florean C,Dicato M.Redox biology of regulated cell death in cancer: a focus on necroptosis and ferroptosis..Free Radic Biol Med2019;134:177-89

[22] Moreno-Caceres J.Apoptosis in liver carcinogenesis and chemotherapy..Hepat Oncol2015;2:381-97 PMCID:PMC6095415

[23] Qu X,Shen L,Xu Y.Autophagy inhibitor chloroquine increases sensitivity to cisplatin in QBC939 cholangiocarcinoma cells by mitochondrial ROS..PLoS One2017;12:e0173712 PMCID:PMC5354635

[24] Guo XL,Sun K,Liu Y.Inhibition of autophagy enhances anticancer effects of bevacizumab in hepatocarcinoma..J Mol Med (Berl)2013;91:473-83 PMCID:PMC3611041

[25] Mizutani H,Nishimoto A,Miyazawa D.Pirarubicin, an anthracycline anticancer agent, induces apoptosis through generation of hydrogen peroxide..Anticancer Res2017;37:6063-9

[26] Dai Y,Huang G,Sheng S.Dichloroacetate enhances adriamycin-induced hepatoma cell toxicity in vitro and in vivo by increasing reactive oxygen species levels..PLoS One2014;9:e92962 PMCID:PMC3984074

[27] Pang Y,Wu L,Chen T.Artesunate induces ROS-dependent apoptosis via a Bax-mediated intrinsic pathway in Huh-7 and Hep3B cells..Exp Cell Res2016;347:251-60

[28] Tang Y,Wang C,Li L.14-3-3zeta binds to hepatitis B virus protein X and maintains its protein stability in hepatocellular carcinoma cells..Cancer Med2018;7:5543-53 PMCID:PMC6247021

[29] Fukuhara T,Ono C,Motooka D.Quasispecies of hepatitis C virus participate in cell-specific infectivity..Sci Rep2017;7:45228 PMCID:PMC5361118

[30] Kambara H,Shiokawa M,Ohara Y.Establishment of a novel permissive cell line for the propagation of hepatitis C virus by expression of microRNA miR122..J Virol2012;86:1382-93 PMCID:PMC3264374

[31] Nayak MK,Doddapattar P,Sonkar VK.Dichloroacetate, an inhibitor of pyruvate dehydrogenase kinases, inhibits platelet aggregation and arterial thrombosis..Blood Adv2018;2:2029-38 PMCID:PMC6093723

[32] Michelakis ED,Mackey JR.Dichloroacetate (DCA) as a potential metabolic-targeting therapy for cancer..Br J Cancer2008;99:6 PMCID:PMC2567082

[33] Khan A,Blackburn AC.Long-term stabilization of stage 4 colon cancer using sodium dichloroacetate therapy..World J Clin Cases2016;4:336-43 PMCID:PMC5067498

[34] Khan A,Blackburn AC.Long-term stabilization of stage 4 colon cancer using sodium dichloroacetate therapy..World J Clin Cases2016;4:8 PMCID:PMC5067498

[35] Stacpoole PW,Barnes C,Carney PR.Controlled clinical trial of dichloroacetate for treatment of congenital lactic acidosis in children..Pediatrics2006;117:12

[36] Abdelmalak M,Ramezani R,Coats BS.Long-term safety of dichloroacetate in congenital lactic acidosis..Mol Genet Metab2013;109:5 PMCID:PMC3751427

[37] Dhanasekaran DN.JNK-signaling: a multiplexing hub in programmed cell death..Genes Cancer2017;8:682-94 PMCID:PMC5724802

[38] Dunnill CJ,Mohamed A,Georgopoulos NT.A redox state-dictated signalling pathway deciphers the malignant cell specificity of CD40-mediated apoptosis..Oncogene2017;36:2515-28 PMCID:PMC5422712

[39] Win S,Kaplowitz N.The regulation of JNK signaling pathways in cell death through the interplay with mitochondrial SAB and upstream post-translational effects..Int J Mol Sci2018;19:3657 PMCID:PMC6274687

[40] Stander XX,Joubert AM.Synergistic anticancer potential of dichloroacetate and estradiol analogue exerting their effect via ROS-JNK-Bcl-2-mediated signalling pathways..Cell Physiol Biochem2015;35:1499-526

[41] Yang X,Wu J,Yan LJ.Humanin attenuates NMDA-induced excitotoxicity by inhibiting ROS-dependent JNK/p38 MAPK pathway..Int J Mol Sci2018;19:2982 PMCID:PMC6213259