Inhibitory effect of metformin combined with gemcitabine on pancreatic cancer cells in vitro and in vivo

Shi,Yuqi; He,Zhilong; Jia,Zhenyu; Xu,Chunfang

doi:10.3892/mmr.2016.5592

Inhibitory effect of metformin combined with gemcitabine on pancreatic cancer cells in vitro and in vivo

Authors:
- Yuqi Shi
- Zhilong He
- Zhenyu Jia
- Chunfang Xu
View Affiliations

Affiliations: Department of Gastroenterology, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu 215006, P.R. China
Published online on: August 4, 2016 https://doi.org/10.3892/mmr.2016.5592
Pages: 2921-2928
Copyright: © Shi 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

Pancreatic cancer is a malignant digestive system tumor with a particularly poor prognosis, and is the fourth leading cause of cancer‑associated mortality in the USA. The anti‑diabetic therapeutic agent, metformin (MET) has been demonstrated to exert anti‑tumor effects. The present study assessed the ability of MET, alone or in combination with gemcitabine (GEM), to inhibit the growth of the human CFPAC‑1 pancreatic cancer cell line in vitro and in vivo. Cell counting kit‑8 assays were performed to measure CFPAC‑1 cell viability and apoptosis was detected with annexin V/propidium iodide. Cell cycle analysis was conducted by flow cytometry. The mRNA and protein levels of B‑cell lymphoma‑extra large (Bcl‑xL), Bcl2 associated X protein (Bax), caspase‑3, survivin and cyclin D1 in CFPAC‑1 cells and tumor tissues were detected by reverse transcription‑polymerase chain reaction and western blotting, respectively. Furthermore, the expression levels of caspase‑3 and proliferating cell nuclear antigen in tumor tissues were detected by immunohistochemistry. The results demonstrated that following MET treatment, the growth of CFPAC‑1 cells and xenografts in nude mice was inhibited, the expression levels of Bcl‑xL, survivin and cyclin D1 were downregulated, while the expression levels of Bax and caspase‑3 were upregulated. These effects were enhanced when MET was administered in combination with GEM. The mechanism underlying the anti‑tumor effect of MET may be associated with the induction of cell apoptosis and the inhibition of proliferation.

View Figures

View References

1	Siegel RL, Miller KD and Jemal A: Cancer statistics, 2015. CA Cancer J Clin. 65:5–29. 2015. View Article : Google Scholar : PubMed/NCBI
2	Xu Q, Zhang TP and Zhao YP: Advances in early diagnosis and therapy of pancreatic cancer. Hepatobiliary Pancreat Dis Int. 10:128–135. 2011. View Article : Google Scholar : PubMed/NCBI
3	Stathis A and Moore MJ: Advanced pancreatic carcinoma: Current treatment and future challenges. Nat Rev Clin Oncol. 7:163–172. 2010. View Article : Google Scholar : PubMed/NCBI
4	Burris HR, Moore MJ, Andersen J, Green MR, Rothenberg ML, Modiano MR, Cripps MC, Portenoy RK, Storniolo AM, Tarassoff P, et al: Improvements in survival and clinical benefit with gemcitabine as first-line therapy for patients with advanced pancreas cancer: A randomized trial. J Clin Oncol. 15:2403–2413. 1997.PubMed/NCBI
5	He H, Ke R, Lin H, Ying Y, Liu D and Luo Z: Metformin, an old drug, brings a new era to cancer therapy. Cancer J. 21:70–74. 2015. View Article : Google Scholar : PubMed/NCBI
6	Morales DR and Morris AD: Metformin in cancer treatment and prevention. Annu Rev Med. 66:17–29. 2015. View Article : Google Scholar
7	Kisfalvi K, Moro A, Sinnett-Smith J, Eibl G and Rozengurt E: Metformin inhibits the growth of human pancreatic cancer xenografts. Pancreas. 42:781–785. 2013. View Article : Google Scholar : PubMed/NCBI
8	Bao B, Wang Z, Ali S, Ahmad A, Azmi AS, Sarkar SH, Banerjee S, Kong D, Li Y, Thakur S and Sarkar FH: Metformin inhibits cell proliferation, migration and invasion by attenuating CSC function mediated by deregulating miRNAs in pancreatic cancer cells. Cancer Prev Res (Phila). 5:355–364. 2012. View Article : Google Scholar
9	Wang LW, Li ZS, Zou DW, Jin ZD, Gao J and Xu GM: Metformin induces apoptosis of pancreatic cancer cells. World J Gastroenterol. 14:7192–7198. 2008. View Article : Google Scholar : PubMed/NCBI
10	Gao ZY, Liu Z, Bi MH, Zhang JJ, Han ZQ, Han X, Wang HY, Sun GP and Liu H: Metformin induces apoptosis via a mitochondria-mediated pathway in human breast cancer cells in vitro. Exp Ther Med. 11:1700–1706. 2016.PubMed/NCBI
11	Fujimori T, Kato K, Fujihara S, Iwama H, Yamashita T, Kobayashi K, Kamada H, Morishita A, Kobara H, Mori H, et al: Antitumor effect of metformin on cholangiocarcinoma: In vitro and in vivo studies. Oncol Rep. 34:2987–2996. 2015.PubMed/NCBI
12	Ramjeesingh R, Orr C, Bricks CS, Hopman WM and Hammad N: A retrospective study on the role of diabetes and metformin in colorectal cancer disease survival. Curr Oncol. 23:e116–e122. 2016. View Article : Google Scholar : PubMed/NCBI
13	Owen MR, Doran E and Halestrap AP: Evidence that metformin exerts its anti-diabetic effects through inhibition of complex 1 of the mitochondrial respiratory chain. Biochem J. 348:607–614. 2000. View Article : Google Scholar : PubMed/NCBI
14	El-Mir MY, Nogueira V, Fontaine E, Averet N, Rigoulet M and Leverve X: Dimetforminhylbiguanide inhibits cell respiration via an indirect effect targeted on the respiratory chain complex I. J Biol Chem. 275:223–228. 2000. View Article : Google Scholar : PubMed/NCBI
15	Gallagher EJ and LeRoith D: Minireview: IGF, insulin, and cancer. Endoscopy. 152:2546–2255. 2011.
16	Algire C, Moiseeva O, Deschênes-Simard X, Amrein L, Petruccelli L, Birman E, Viollet B, Ferbeyre G and Pollak MN: Metformin reduces endogenous reactive oxygen species and associated DNA damage. Cancer Prev Res (Phila). 5:536–543. 2012. View Article : Google Scholar
17	Hirsch HA, Iliopoulos D and Struhl K: Metformin inhibits the inflammatory response associated with cellular transformation and cancer stem cell growth. Proc Natl Acad Sci USA. 110:972–977. 2013. View Article : Google Scholar : PubMed/NCBI
18	Gou S, Cui P, Li X, Shi P, Liu T and Wang C: Low concentrations of metformin selectively inhibit CD133 (+) cell proliferation in pancreatic cancer and have anticancer action. Plos One. 8:e639692013. View Article : Google Scholar
19	Breckenridge DG and Xue D: Regulation of mitochondrial membrane permeabilization by BCL-2 family proteins and caspases. Curr Opin Cell Biol. 16:647–652. 2004. View Article : Google Scholar : PubMed/NCBI
20	Yang J, Liu X, Bhalla K, Kim CN, Ibrado AM, Cai J, Peng TI, Jones DP and Wang X: Prevention of apoptosis by Bcl-2: Release of cytochrome c from mitochondria blocked. Science. 275:1129–1132. 1997. View Article : Google Scholar : PubMed/NCBI
21	Amundson SA, Myers TG, Scudiero D, Kitada S, Reed JC and Fornace AJ Jr: An informatics approach identifying markers of chemosensitivity in human cancer cell lines. Cancer Res. 60:6101–6610. 2000.PubMed/NCBI
22	Mobahat M, Narendran A and Riabowol K: Survivin as a preferential target for cancer therapy. Int J Mol Sci. 15:2494–2516. 2014. View Article : Google Scholar : PubMed/NCBI
23	Hassan M, Watari H, AbuAlmaaty A, Ohba Y and Sakuragi N: Apoptosis and molecular targeting therapy in cancer. Biomed Res Int. 2014:1508452014. View Article : Google Scholar : PubMed/NCBI
24	Kim JK and Diehl JA: Nuclear cyclin D1: An oncogenic driver in human cancer. J Cell Physiol. 220:292–296. 2009. View Article : Google Scholar : PubMed/NCBI
25	Juríková M, Danihel Ľ, Polák Š and Varga I: Ki67, PCNA, and MCM proteins: Markers of proliferation in the diagnosis of breast cancer. Acta Histochem. 30084–30088. 2016.Epub ahead of print.
26	Vincent A, Herman J, Schulick R, Hruban RH and Goggins M: Pancreatic cancer. Lancet. 378:607–620. 2011. View Article : Google Scholar : PubMed/NCBI
27	Colucci G, Labianca R, Di Costanzo F, Gebbia V, Cartenì G, Massidda B, Dapretto E, Manzione L, Piazza E, Sannicolò M, et al: Randomized phase III trial of gemcitabine plus cisplatin compared with single-agent gemcitabine as first-line treatment of patients with advanced pancreatic cancer: The GIP-1 study. J Clin Oncol. 28:1645–1651. 2010. View Article : Google Scholar : PubMed/NCBI
28	Abou-Alfa GK, Letourneau R, Harker G, Modiano M, Hurwitz H, Tchekmedyian NS, Feit K, Ackerman J, De Jager RL, Eckhardt SG and O'Reilly EM: Randomized phase III study of exatecan and gemcitabine compared with gemcitabine alone in untreated advanced pancreatic cancer. J Clin Oncol. 24:4441–4447. 2006. View Article : Google Scholar : PubMed/NCBI
29	Cunningham D, Chau I, Stocken DD, Valle JW, Smith D, Steward W, Harper PG, Dunn J, Tudur-Smith C, West J, et al: Phase III randomized comparison of gemcitabine versus gemcitabine plus capecitabine in patients with advanced pancreatic cancer. J Clin Oncol. 27:5513–5518. 2009. View Article : Google Scholar : PubMed/NCBI
30	Gong J, Robbins LA, Lugea A, Waldron RT, Jeon CY and Pandol SJ: Diabetes, pancreatic cancer, and metformin therapy. Front Physiol. 5:4262014. View Article : Google Scholar : PubMed/NCBI
31	Choi Y, Kim TY, Oh DY, Lee KH, Han SW, Im SA, Kim TY and Bang YJ: The impact of diabetes mellitus and metformin treatment on survival of patients with advanced pancreatic cancer undergoing chemotherapy. Cancer Res Treat. 48:171–179. 2016. View Article : Google Scholar :
32	Sadeghi N, Abbruzzese JL, Yeung SC, Hassan M and Li D: Metformin use is associated with better survival of diabetic patients with pancreatic cancer. Clin Cancer Res. 18:2905–2912. 2012. View Article : Google Scholar : PubMed/NCBI
33	Zhang JW and Sun Q: Metformin may improve the prognosis of patients with pancreatic cancer. Asian Pac J Cancer Prev. 16:3937–3940. 2015. View Article : Google Scholar : PubMed/NCBI
34	Kordes S, Pollak MN, Zwinderman AH, Mathôt RA, Weterman MJ, Beeker A, Punt CJ, Richel DJ and Wilmink JW: Metformin in patients with advanced pancreatic cancer: A double-blind, randomised, placebo-controlled phase 2 trial. Lancet Oncol. 16:839–847. 2015. View Article : Google Scholar : PubMed/NCBI