BH3 profiling discriminates the anti‑apoptotic status of 5‑fluorouracil‑resistant colon cancer cells

Ishikawa,Kazuma; Kawano,Yutaka; Arihara,Yohei; Kubo,Tomohiro; Takada,Kohichi; Murase,Kazuyuki; Miyanishi,Koji; Kobune,Masayoshi; Kato,Junji

doi:10.3892/or.2019.7373

BH3 profiling discriminates the anti‑apoptotic status of 5‑fluorouracil‑resistant colon cancer cells

Authors:
- Kazuma Ishikawa
- Yutaka Kawano
- Yohei Arihara
- Tomohiro Kubo
- Kohichi Takada
- Kazuyuki Murase
- Koji Miyanishi
- Masayoshi Kobune
- Junji Kato
View Affiliations

Affiliations: Department of Medical Oncology, Sapporo Medical University School of Medicine, Sapporo, Hokkaido 060‑8543, Japan, Department of Hematology, Sapporo Medical University School of Medicine, Sapporo, Hokkaido 060‑8543, Japan
Published online on: October 15, 2019 https://doi.org/10.3892/or.2019.7373
Pages: 2416-2425
Copyright : © Ishikawa et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY 4.0].

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) is a cytotoxic anticancer drug commonly used for patients with advanced colon cancer. This drug effectively reduces the size of tumors to a certain degree; however, cancer cells can gradually acquire resistance, resulting in disease progression. To identify the mechanism of 5‑FU resistance, we established three 5‑FU‑resistant colon cancer cell lines and analyzed both apoptosis‑related protein expression levels and BH3 profiling. These 5‑FU‑resistant colon cancer cell lines acquired apoptotic resistance to 5‑FU. Although apoptosis‑related protein expression levels were altered in each 5‑FU‑resistant colon cancer cell line variably, BH3 profiling indicated BCLXL dependence in 5‑FU‑resistant HT‑29 cells only. Functional BCLXL inhibition in 5‑FU‑resistant HT‑29 cells not only sensitized the cells to apoptosis but also overcame 5‑FU resistance. The apoptotic BIM protein was preferentially sequestered, thereby resulting in acquired dependence on BCLXL for survival. Additionally, in vivo models showed that BCLXL inhibition controlled tumor progression. These results indicate that BH3 profiling facilitates the identification of the functional role of anti‑apoptotic proteins during drug resistance and has clinical implications for colon cancer in targeting specific proteins such as BCLXL.

View Figures

View References

1	Arnold M, Sierra MS, Laversanne M, Soerjomataram I, Jemal A and Bray F: Global patterns and trends in colorectal cancer incidence and mortality. Gut. 66:683–691. 2017. View Article : Google Scholar : PubMed/NCBI
2	Hori M, Matsuda T, Shibata A, Katanoda K, Sobue T and Nishimoto H; Japan Cancer Surveillance Research Group, : Cancer incidence and incidence rates in Japan in 2009: A study of 32 population-based cancer registries for the Monitoring of Cancer Incidence in Japan (MCIJ) project. Jpn J Clin Oncol. 45:884–891. 2015. View Article : Google Scholar : PubMed/NCBI
3	Watanabe T, Muro K, Ajioka Y, Hashiguchi Y, Ito Y, Saito Y, Hamaguchi T, Ishida H, Ishiguro M, Ishihara S, et al: Japanese society for cancer of the colon and rectum (JSCCR) guidelines 2016 for the treatment of colorectal cancer. Int J Clin Oncol. 23:1–34. 2018. View Article : Google Scholar : PubMed/NCBI
4	Rutman RJ, Cantarow A and Paschkis KE: Studies in 2-acetylaminofluorene carcinogenesis. III. The utilization of uracil-2-C14 by preneoplastic rat liver and rat hepatoma. Cancer Res. 14:119–123. 1954.PubMed/NCBI
5	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
6	Liu W, Fang Y, Wang XT, Liu J, Dan X and Sun LL: Overcoming 5-Fu resistance of colon cells through inhibition of Glut1 by the specific inhibitor WZB117. Asian Pac J Cancer Prev. 15:7037–7041. 2014. View Article : Google Scholar : PubMed/NCBI
7	Dallas NA, Xia L, Fan F, Gray MJ, Gaur P, van Buren G II, Samuel S, Kim MP, Lim SJ and Ellis LM: Chemoresistant colorectal cancer cells, the cancer stem cell phenotype, and increased sensitivity to insulin-like growth factor-I receptor inhibition. Cancer Res. 69:1951–1957. 2009. View Article : Google Scholar : PubMed/NCBI
8	Yoshida M, Horiguchi H, Kikuchi S, Iyama S, Ikeda H, Goto A, Kawano Y, Murase K, Takada K, Miyanishi K, et al: miR-7977 inhibits the Hippo-YAP signaling pathway in the bone marrow mesenchymal stromal cells. PLoS One. 14:e02132202019. View Article : Google Scholar : PubMed/NCBI
9	Ryan J and Letai A: BH3 profiling in whole cells by fluorimeter or FACS. Methods. 61:156–164. 2013. View Article : Google Scholar : PubMed/NCBI
10	Fraser C, Ryan J and Sarosiek K: BH3 profiling: A functional assay to measure apoptotic priming and dependencies. Methods Mol Biol. 1877:61–76. 2019. View Article : Google Scholar : PubMed/NCBI
11	Tao ZF, Hasvold L, Wang L, Wang X, Petros AM, Park CH, Boghaert ER, Catron ND, Chen J, Colman PM, et al: Discovery of a potent and selective BCL-XL inhibitor with in vivo activity. ACS Med Chem Lett. 5:1088–1093. 2014. View Article : Google Scholar : PubMed/NCBI
12	Jian YS, Chen CW, Lin CA, Yu HP, Lin HY, Liao MY, Wu SH, Lin YF and Lai PS: Hyaluronic acid-nimesulide conjugates as anticancer drugs against CD44-overexpressing HT-29 colorectal cancer in vitro and in vivo. Int J Nanomedicine. 12:2315–2333. 2017. View Article : Google Scholar : PubMed/NCBI
13	Dutta S, Ryan J, Chen TS, Kougentakis C, Letai A and Keating AE: Potent and specific peptide inhibitors of human pro-survival protein Bcl-xL. J Mol Biol. 427:1241–1253. 2015. View Article : Google Scholar : PubMed/NCBI
14	Foight GW, Ryan JA, Gullá SV, Leta A and Keating AE: Designed BH3 peptides with high affinity and specificity for targeting Mcl-1 in cells. ACS Chem Biol. 9:1962–1968. 2014. View Article : Google Scholar : PubMed/NCBI
15	Montero J and Letai A: Why do BCL-2 inhibitors work and where should we use them in the clinic? Cell Death Differ. 25:56–64. 2018. View Article : Google Scholar : PubMed/NCBI
16	Nita ME, Nagawa H, Tominaga O, Tsuno N, Fujii S, Sasaki S, Fu CG, Takenoue T, Tsuruo T and Muto T: 5-Fluorouracil induces apoptosis in human colon cancer cell lines with modulation of Bcl-2 family proteins. Br J Cancer. 78:986–992. 1998. View Article : Google Scholar : PubMed/NCBI
17	Zhao DP, Ding XW, Peng JP, Zheng YX and Zhang SZ: Prognostic significance of bcl-2 and p53 expression in colorectal carcinoma. J Zhejiang Univ Sci B. 6:1163–1169. 2005. View Article : Google Scholar : PubMed/NCBI
18	Wu DW, Huang CC, Chang SW, Chen TH and Lee H: Bcl-2 stabilization by paxillin confers 5-fluorouracil resistance in colorectal cancer. Cell Death Differ. 22:779–789. 2015. View Article : Google Scholar : PubMed/NCBI
19	Vo TT, Ryan J, Carrasco R, Neuberg D, Rossi DJ, Stone RM, Deangelo DJ, Frattini MG and Letai A: Relative mitochondrial priming of myeloblasts and normal HSCs determines chemotherapeutic success in AML. Cell. 151:344–355. 2012. View Article : Google Scholar : PubMed/NCBI
20	Ni Chonghaile T, Sarosiek KA, Vo TT, Ryan JA, Tammareddi A, Moore Vdel G, Deng J, Anderson KC, Richardson P, Tai YT, et al: Pretreatment mitochondrial priming correlates with clinical response to cytotoxic chemotherapy. Science. 334:1129–1133. 2011. View Article : Google Scholar : PubMed/NCBI
21	Kubo T, Kawano Y, Himuro N, Sugita S, Sato Y, Ishikawa K, Takada K, Murase K, Miyanishi K, Sato T, et al: BAK is a predictive and prognostic biomarker for the therapeutic effect of docetaxel treatment in patients with advanced gastric cancer. Gastric Cancer. 19:827–838. 2016. View Article : Google Scholar : PubMed/NCBI
22	Klotz DM, Nelson SA, Kroboth K, Newton IP, Radulescu S, Ridgway RA, Sansom OJ, Appleton PL and Näthke IS: The microtubule poison vinorelbine kills cells independently of mitotic arrest and targets cells lacking the APC tumour suppressor more effectively. J Cell Sci. 125:887–895. 2012. View Article : Google Scholar : PubMed/NCBI
23	Yan Y, Xie R, Zhang Q, Zhu X, Han J and Xia R: Bcl-xL/Bak interaction and regulation by miRNA let-7b in the intrinsic apoptotic pathway of stored platelets. Platelets. 30:75–80. 2019. View Article : Google Scholar : PubMed/NCBI