Bufalin and 5‑fluorouracil synergistically induce apoptosis in colorectal cancer cells

Dai,Xiao‑Yu; Zhou,Bao‑Feng; Xie,Yang‑Yang; Lou,Jie; Li,Ke‑Qiang

doi:10.3892/ol.2018.8332

Bufalin and 5‑fluorouracil synergistically induce apoptosis in colorectal cancer cells

Authors:
- Xiao‑Yu Dai
- Bao‑Feng Zhou
- Yang‑Yang Xie
- Jie Lou
- Ke‑Qiang Li
View Affiliations

Affiliations: Department of Anorectal Surgery, Clinical Research Center, Ningbo No. 2 Hospital, Ningbo, Zhejiang 315010, P.R. China, Department of Digestion, Clinical Research Center, Ningbo No. 2 Hospital, Ningbo, Zhejiang 315010, P.R. China, Key Laboratory of Molecular Biology of Cancer, Clinical Research Center, Ningbo No. 2 Hospital, Ningbo, Zhejiang 315010, P.R. China
Published online on: March 23, 2018 https://doi.org/10.3892/ol.2018.8332
Pages: 8019-8026

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

5‑fluorouracil (5‑FU) has been used in the treatment of colorectal cancer for >50 years. However, drug resistance remains an obstacle in the application of 5‑FU‑based chemotherapy. Bufalin, a type of steroid with anti‑tumor activity, may be purified from the skin and parotid venom glands of toads. In order to improve the anti‑tumor effect of 5‑FU, the present study examined the combined effects of bufalin with 5‑FU on human colorectal cancer HCT116 cells. Following treatment, cell proliferation was quantified using MTT assay and apoptotic cell percentage was assessed by flow cytometry. The apoptosis‑associated protein expression was evaluated by western blotting. It was observed that bufalin enhanced the cytotoxicity of 5‑FU in HCT116 cells via the induction of the mitochondrial apoptotic pathway. Additionally, bufalin combined with 5‑FU reduced the expression levels of anti‑apoptotic proteins, such as Mcl‑1, XIAP and Bcl‑2 and upregulated the levels of the pro‑apoptotic proteins, Bax and Bad. To verify the role of Bax, RNA interference was used to knock‑down Bax. It was determined that the synergistic effect between 5‑FU and bufalin was diminished following the silencing of Bax. In summary, bufalin in combination with 5‑FU may induce a higher level of apoptosis compared with monotherapy, and the combination mat be a potential therapeutic strategy for the treatment of colorectal cancer.

View Figures

View References

1	Siegel RL, Miller KD and Jemal A: Cancer statistics, 2016. CA Cancer J Clin. 66:7–30. 2016. View Article : Google Scholar : PubMed/NCBI
2	Torre LA, Siegel RL, Ward EM and Jemal A: Global cancer incidence and mortality rates and trends-an update. Cancer Epidemiol Biomarkers Prev. 25:16–27. 2016. View Article : Google Scholar : PubMed/NCBI
3	Mayer RJ: Targeted therapy for advanced colorectal cancer-more is not always better. N Engl J Med. 360:623–625. 2009. View Article : Google Scholar : PubMed/NCBI
4	Longley DB, Harkin DP and Johnston PG: 5-fluorouracil: Mechanisms of action and clinical strategies. Nat Rev Cancer. 3:330–338. 2003. View Article : Google Scholar : PubMed/NCBI
5	Temraz S, Mukherji D, Alameddine R and Shamseddine A: Methods of overcoming treatment resistance in colorectal cancer. Crit Rev Oncol Hematol. 89:217–230. 2014. View Article : Google Scholar : PubMed/NCBI
6	Yu R, Deedigan L, Albarenque SM, Mohr A and Zwacka RM: Delivery of sTRAIL variants by MSCs in combination with cytotoxic drug treatment leads to p53-independent enhanced antitumor effects. Cell Death Dis. 4:e5032013. View Article : Google Scholar : PubMed/NCBI
7	Rich JN and Bao S: Chemotherapy and cancer stem cells. Cell Stem Cell. 1:353–355. 2007. View Article : Google Scholar : PubMed/NCBI
8	Tummers B and Green DR: Caspase-8: Regulating life and death. Immunol Rev. 277:76–89. 2017. View Article : Google Scholar : PubMed/NCBI
9	Estaquier J, Vallette F, Vayssiere JL and Mignotte B: The mitochondrial pathways of apoptosis. Adv Exp Med Biol. 942:157–183. 2012. View Article : Google Scholar : PubMed/NCBI
10	Susin SA, Lorenzo HK, Zamzami N, Marzo I, Snow BE, Brothers GM, Mangion J, Jacotot E, Costantini P, Loeffler M, et al: Molecular characterization of mitochondrial apoptosis-inducing factor. Nature. 397:441–446. 1999. View Article : Google Scholar : PubMed/NCBI
11	Azmi AS, Wang Z, Philip PA, Mohammad RM and Sarkar FH: Emerging Bcl-2 inhibitors for the treatment of cancer. Expert Opin Emerg Drugs. 16:59–70. 2011. View Article : Google Scholar : PubMed/NCBI
12	Shakibaei M, Mobasheri A, Lueders C, Busch F, Shayan P and Goel A: Curcumin enhances the effect of chemotherapy against colorectal cancer cells by inhibition of NF-κB and Src protein kinase signaling pathways. PLoS One. 8:e572182013. View Article : Google Scholar : PubMed/NCBI
13	Douillard JY, Sobrero A, Carnaghi C, Comella P, Díaz-Rubio E, Santoro A and Van Cutsem E: Metastatic colorectal cancer: Integrating irinotecan into combination and sequential chemotherapy. Ann Oncol. 14 Suppl 2:ii7–ii12. 2003. View Article : Google Scholar : PubMed/NCBI
14	Giacchetti S, Perpoint B, Zidani R, Le Bail N, Faggiuolo R, Focan C, Chollet P, Llory JF, Letourneau Y, Coudert B, et al: Phase III multicenter randomized trial of oxaliplatin added to chronomodulated fluorouracil-leucovorin as first-line treatment of metastatic colorectal cancer. J Clin Oncol. 18:136–147. 2000. View Article : Google Scholar : PubMed/NCBI
15	Lu CX, Nan KJ and Lei Y: Agents from amphibians with anticancer properties. Anticancer Drugs. 19:931–939. 2008. View Article : Google Scholar : PubMed/NCBI
16	Miao Q, Bi LL, Li X, Miao S, Zhang J, Zhang S, Yang Q, Xie YH, Zhang J and Wang SW: Anticancer effects of bufalin on human hepatocellular carcinoma HepG2 cells: Roles of apoptosis and autophagy. Int J Mol Sci. 14:1370–1382. 2013. View Article : Google Scholar : PubMed/NCBI
17	Xie CM, Chan WY, Yu S, Zhao J and Cheng CH: Bufalin induces autophagy-mediated cell death in human colon cancer cells through reactive oxygen species generation and JNK activation. Free Radic Biol Med. 51:1365–1375. 2011. View Article : Google Scholar : PubMed/NCBI
18	Santandreu FM, Valle A, Oliver J and Roca P: Resveratrol potentiates the cytotoxic oxidative stress induced by chemotherapy in human colon cancer cells. Cell Physiol Biochem. 28:219–228. 2011. View Article : Google Scholar : PubMed/NCBI
19	Fishbein AB, Wang CZ, Li XL, Mehendale SR, Sun S, Aung HH and Yuan CS: Asian ginseng enhances the anti-proliferative effect of 5-fluorouracil on human colorectal cancer: Comparison between white and red ginseng. Arch Pharm Res. 32:505–513. 2009. View Article : Google Scholar : PubMed/NCBI
20	Jiang L, Zhao MN, Liu TY, Wu XS, Weng H, Ding Q, Shu YJ, Bao RF, Li ML, Mu JS, et al: Bufalin induces cell cycle arrest and apoptosis in gallbladder carcinoma cells. Tumour Biol. 35:10931–10941. 2014. View Article : Google Scholar : PubMed/NCBI
21	Qiu DZ, Zhang ZJ, Wu WZ and Yang YK: Bufalin, a component in Chansu, inhibits proliferation and invasion of hepatocellular carcinoma cells. BMC Complement Altern Med. 13:1852013. View Article : Google Scholar : PubMed/NCBI
22	Takai N, Ueda T, Nishida M, Nasu K and Narahara H: Bufalin induces growth inhibition, cell cycle arrest and apoptosis in human endometrial and ovarian cancer cells. Int J Mol Med. 21:637–643. 2008.PubMed/NCBI
23	Chang Y, Zhao Y, Gu W, Cao Y, Wang S, Pang J and Shi Y: Bufalin inhibits the differentiation and proliferation of cancer stem cells derived from primary osteosarcoma cells through Mir-148a. Cell Physiol Biochem. 36:1186–1196. 2015. View Article : Google Scholar : PubMed/NCBI
24	Zhao L, Liu S, Che X, Hou K, Ma Y, Li C, Wen T, Fan Y, Hu X, Liu Y and Qu X: Bufalin inhibits TGF-β-induced epithelial-to-mesenchymal transition and migration in human lung cancer A549 cells by downregulating TGF-β receptors. Int J Mol Med. 36:645–652. 2015. View Article : Google Scholar : PubMed/NCBI
25	Sampath D, Rao VA and Plunkett W: Mechanisms of apoptosis induction by nucleoside analogs. Oncogene. 22:9063–9074. 2003. View Article : Google Scholar : PubMed/NCBI
26	Hong SH and Choi YH: Bufalin induces apoptosis through activation of both the intrinsic and extrinsic pathways in human bladder cancer cells. Oncol Rep. 27:114–120. 2012.PubMed/NCBI
27	Datta SR, Katsov A, Hu L, Petros A, Fesik SW, Yaffe MB and Greenberg ME: 14-3-3 proteins and survival kinases cooperate to inactivate BAD by BH3 domain phosphorylation. Mol Cell. 6:41–51. 2000. View Article : Google Scholar : PubMed/NCBI
28	Qu Y, Xia P, Zhang S, Pan S and Zhao J: Silencing XIAP suppresses osteosarcoma cell growth, and enhances the sensitivity of osteosarcoma cells to doxorubicin and cisplatin. Oncol Rep. 33:1177–1184. 2015. View Article : Google Scholar : PubMed/NCBI
29	Mehrotra S, Languino LR, Raskett CM, Mercurio AM, Dohi T and Altieri DC: IAP regulation of metastasis. Cancer Cell. 17:53–64. 2010. View Article : Google Scholar : PubMed/NCBI
30	Qi F, Li A, Inagaki Y, Kokudo N, Tamura S, Nakata M and Tang W: Antitumor activity of extracts and compounds from the skin of the toad Bufo bufo gargarizans Cantor. Int Immunopharmacol. 11:342–349. 2011. View Article : Google Scholar : PubMed/NCBI
31	Meng Z, Yang P, Shen Y, Bei W, Zhang Y, Ge Y, Newman RA, Cohen L, Liu L, Thornton B, et al: Pilot study of huachansu in patients with hepatocellular carcinoma, nonsmall-cell lung cancer, or pancreatic cancer. Cancer. 115:5309–5318. 2009. View Article : Google Scholar : PubMed/NCBI