NaHCO3 enhances the antitumor activities of cytokine-induced killer cells against hepatocellular carcinoma HepG2 cells

Yuan,Ya  Hong; Zhou,Chun  Fang; Yuan,Jiang; Liu,Li; Guo,Xing  Rong; Wang,Xiao  Li; Ding,Yan; Wang,Xiao  Nan; Li,Dong  Sheng; Tu,Han  Jun

doi:10.3892/ol.2016.5112

NaHCO3 enhances the antitumor activities of cytokine-induced killer cells against hepatocellular carcinoma HepG2 cells

Authors:
- Ya Hong Yuan
- Chun Fang Zhou
- Jiang Yuan
- Li Liu
- Xing Rong Guo
- Xiao Li Wang
- Yan Ding
- Xiao Nan Wang
- Dong Sheng Li
- Han Jun Tu
View Affiliations

Affiliations: Hubei Key Laboratory of Embryonic Stem Cell Research, Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei 442000, P.R. China, Department of Gastroenterology, Renmin Hospital, Hubei University of Medicine, Shiyan, Hubei 442000, P.R. China, Department of Neurology, Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei 442000, P.R. China, Department of Infectious Diseases, Tianyou Hospital, Wuhan University of Science and Technology, Wuhan, Hubei 430000, P.R. China
Published online on: September 9, 2016 https://doi.org/10.3892/ol.2016.5112
Pages: 3167-3174
Copyright: © Yuan 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

The extracellular pH is lower inside solid tumors than in normal tissue. The acidic environment inhibits the cytotoxicity of lymphocytes in vitro and promotes tumor cell invasion. In the present study, both in vitro and in vivo experiments were conducted to investigate how NaHCO3 would affect the antitumor activities of cytokine‑induced killer (CIK) cells against hepatocellular carcinoma (HCC) cells. For the in vitro experiments, HepG2 cells were cultured at pH 6.5 and 7.4 in the presence of CIK cells or CIK cell‑conditioned medium (CMCIK). For the in vivo experiments, nude mice were xenografted with HepG2‑luc cells and divided into four groups: i) CIK cells injection plus NaHCO3 feeding; ii) CIK cells injection plus drinking water feeding; iii) normal saline injection plus NaHCO3 feeding; and iv) normal saline injection plus drinking water feeding. The results indicated that the viability and growth rate of HepG2 cells were remarkably suppressed when co‑cultured with CIK cells or CMCIK at pH 7.4 compared with those of HepG2 cells cultured under the same conditions but at pH 6.5. In the xenograft study, a marked synergistic antitumor effect of the combined therapy was observed. NaHCO3 feeding augmented the infiltration of cluster of differentiation 3‑positive T lymphocytes into the tumor mass. Taken together, these data strongly suggest that the antitumor activities of CIK cells against HepG2 cells were negatively affected by the acidic environment inside the tumors, and neutralizing the pH (for example, via NaHCO3 administration), could therefore reduce or eliminate this influence. In addition, it should be recommended that oncologists routinely prescribe soda water to their patients, particularly during CIK cell therapy.

View Figures

View References

1	El-Serag HB: Epidemiology of viral hepatitis and hepatocellular carcinoma. Gastroenterology. 142:1264.e1–1273.e1. 2012. View Article : Google Scholar
2	Su Y, Yang Y, Ma Y, Zhang Y, Rao W, Yang G and Kou C: The Efficacy and Safety of Dendritic Cells Co-Cultured with Cytokine-Induced Killer Cell Therapy in Combination with TACE-Predominant Minimally-Invasive Treatment for Hepatocellular Carcinoma: A Meta-Analysis. Clin Lab. 62:599–608. 2016.PubMed/NCBI
3	Zhang L, Zhu W, Li J, Yang X, Ren Y, Niu J and Pang Y: Clinical outcome of immunotherapy with dendritic cell vaccine and cytokine-induced killer cell therapy in hepatobiliary and pancreatic cancer. Mol Clin Oncol. 4:129–133. 2016.PubMed/NCBI
4	Schmidt-Wolf IG, Finke S, Trojaneck B, Denkena A, Lefterova P, Schwella N, Heuft HG, Prange G, Korte M, Takeya M, et al: Phase I clinical study applying autologous immunological effector cells transfected with the interleukin-2 gene in patients with metastatic renal cancer, colorectal cancer and lymphoma. Br J Cancer. 81:1009–1016. 1999. View Article : Google Scholar : PubMed/NCBI
5	Rettinger E, Meyer V, Kreyenberg H, Volk A, Kuçi S, Willasch A, Koscielniak E, Fulda S, Wels WS, Boenig H, et al: Cytotoxic capacity of IL-15-stimulated cytokine-induced killer cells against human acute myeloid leukemia and rhabdomyosarcoma in humanized preclinical mouse models. Front Oncol. 2:322012. View Article : Google Scholar : PubMed/NCBI
6	Huang X, Chen YT, Song HZ, Huang GC and Chen LB: Cisplatin pretreatment enhances antitumor activity of cytokine-induced killer cells. World J Gastroenterol. 17:3002–3011. 2011. View Article : Google Scholar : PubMed/NCBI
7	Wang D, Zhang B, Gao H, Ding G, Wu Q, Zhang J, Liao L and Chen H: Clinical research of genetically modified dendritic cells in combination with cytokine-induced killer cell treatment in advanced renal cancer. BMC Cancer. 14:2512014. View Article : Google Scholar : PubMed/NCBI
8	Xie F, Zhang X, Li H, Zheng T, Xu F, Shen R, Yan L, Yang J and He J: Adoptive immunotherapy in postoperative hepatocellular carcinoma: A systemic review. PLoS One. 7:e428792012. View Article : Google Scholar : PubMed/NCBI
9	Cantor JR and Sabatini DM: Cancer cell metabolism: One hallmark, many faces. Cancer Discov. 2:881–898. 2012. View Article : Google Scholar : PubMed/NCBI
10	Andersen AP, Moreira JM and Pedersen SF: Interactions of ion transporters and channels with cancer cell metabolism and the tumour microenvironment. Philos Trans R Soc Lond B Biol Sci. 369:201300982014. View Article : Google Scholar : PubMed/NCBI
11	Gallagher FA, Kettunen MI, Day SE, Hu DE, Ardenkjaer-Larsen JH, Zandt RI, Jensen PR, Karlsson M, Golman K, Lerche MH and Brindle KM: Magnetic resonance imaging of pH in vivo using hyperpolarized 13C-labelled bicarbonate. Nature. 453:940–943. 2008. View Article : Google Scholar : PubMed/NCBI
12	Rofstad EK, Mathiesen B, Kindem K and Galappathi K: Acidic extracellular pH promotes experimental metastasis of human melanoma cells in athymic nude mice. Cancer Res. 66:6699–6707. 2006. View Article : Google Scholar : PubMed/NCBI
13	Barar J and Omidi Y: Dysregulated pH in tumor microenvironment checkmates cancer therapy. Bioimpacts. 3:149–162. 2013.PubMed/NCBI
14	von Schwarzenberg K, Lajtos T, Simon L, Müller R, Vereb G and Vollmar AM: V-ATPase inhibition overcomes trastuzumab resistance in breast cancer. Mol Oncol. 8:9–19. 2014. View Article : Google Scholar : PubMed/NCBI
15	Ferrari M, Fornasiero MC and Isetta AM: MTT colorimetric assay for testing macrophage cytotoxic activity in vitro. J Immunol Methods. 131:165–172. 1990. View Article : Google Scholar : PubMed/NCBI
16	Pyo KH, Jung BK, Xin CF, Lee YW, Chai JY and Shin EH: Prominent IL-12 production and tumor reduction in athymic nude mice after Toxoplasma gondii lysate antigen treatment. Korean J Parasitol. 52:605–612. 2014. View Article : Google Scholar : PubMed/NCBI
17	Chen JH, Xiang JY, Ding GP and Cao LP: Cholangiocarcinoma-derived exosomes inhibit the antitumor activity of cytokine-induced killer cells by down-regulating the secretion of tumor necrosis factor-α and perforin. J Zhejiang Univ Sci B. 17:537–544. 2016. View Article : Google Scholar : PubMed/NCBI
18	Lardner A: The effects of extracellular pH on immune function. J Leukoc Biol. 69:522–530. 2001.PubMed/NCBI
19	Rabinovich M, DeStefano MJ and Dziezanowski MA: Neutrophil migration under agarose: Stimulation by lowered medium pH and osmolality. J Reticuloendothel Soc. 27:189–200. 1980.PubMed/NCBI
20	Leblebicioglu B, Lim JS, Cario AC, Beck FM and Walters JD: pH changes observed in the inflamed gingival crevice modulate human polymorphonuclear leukocyte activation in vitro. J Periodontol. 67:472–477. 1996. View Article : Google Scholar : PubMed/NCBI
21	Loeffler DA, Juneau PL and Masserant S: Influence of tumour physico-chemical conditions on interleukin-2-stimulated lymphocyte proliferation. Br J Cancer. 66:619–622. 1992. View Article : Google Scholar : PubMed/NCBI
22	Kellum JA, Song M and Li J: Science review: Extracellular acidosis and the immune response: Clinical and physiologic implications. Crit Care. 8:331–336. 2004. View Article : Google Scholar : PubMed/NCBI
23	Kraus M and Wolf B: Implications of acidic tumor microenvironment for neoplastic growth and cancer treatment: A computer analysis. Tumour Biol. 17:133–154. 1996. View Article : Google Scholar : PubMed/NCBI
24	Nishimura R, Baker J, Beilhack A, Zeiser R, Olson JA, Sega EI, Karimi M and Negrin RS: In vivo trafficking and survival of cytokine-induced killer cells resulting in minimal GVHD with retention of antitumor activity. Blood. 112:2563–2574. 2008. View Article : Google Scholar : PubMed/NCBI
25	Pievani A, Borleri G, Pende D, Moretta L, Rambaldi A, Golay J and Introna M: Dual-functional capability of CD3+CD56+CIK cells, a T-cell subset that acquires NK function and retains TCR-mediated specific cytotoxicity. Blood. 118:3301–3310. 2011. View Article : Google Scholar : PubMed/NCBI
26	Miao L, Run-Ming J and Yi J: T-Bet mediated anti-neoplastic effects of dendritic cell-cytokine induced killer cells in vitro. Iran J Pediatr. 22:43–51. 2012.PubMed/NCBI
27	Martin NK, Gaffney EA, Gatenby RA, Gillies RJ, Robey IF and Maini PK: A mathematical model of tumour and blood pHe regulation: The HCO3-/CO2 buffering system. Math Biosci. 230:1–11. 2011. View Article : Google Scholar : PubMed/NCBI
28	Robey IF, Baggett BK, Kirkpatrick ND, Roe DJ, Dosescu J, Sloane BF, Hashim AI, Morse DL, Raghunand N, Gatenby RA and Gillies RJ: Bicarbonate increases tumor pH and inhibits spontaneous metastases. Cancer Res. 69:2260–2268. 2009. View Article : Google Scholar : PubMed/NCBI
29	Wojtkowiak JW, Verduzco D, Schramm KJ and Gillies RJ: Drug resistance and cellular adaptation to tumor acidic pH microenvironment. Mol Pharm. 8:2032–2038. 2011. View Article : Google Scholar : PubMed/NCBI
30	Estrella V, Chen T, Lloyd M, Wojtkowiak J, Cornnell HH, Ibrahim-Hashim A, Bailey K, Balagurunathan Y, Rothberg JM, Sloane BF, et al: Acidity generated by the tumor microenvironment drives local invasion. Cancer Res. 73:1524–1535. 2013. View Article : Google Scholar : PubMed/NCBI
31	Silva AS, Yunes JA, Gillies RJ and Gatenby RA: The potential role of systemic buffers in reducing intratumoral extracellular pH and acid-mediated invasion. Cancer Res. 69:2677–2684. 2009. View Article : Google Scholar : PubMed/NCBI
32	Robey IF and Nesbit LA: Investigating mechanisms of alkalinization for reducing primary breast tumor invasion. Biomed Res Int. 2013:4851962013. View Article : Google Scholar : PubMed/NCBI
33	Chu H, Du F, Jiang L, Wang Z, Gong Z, Lian P, Li P and Chen J: The Efficacy of CIK-Based Immunotherapies for Advanced Solid Tumors. Technol Cancer Res Treat. Jul 19–2016.(Epub ahead of print).