5-FU preferably induces apoptosis in BRAF V600E colorectal cancer cells via downregulation of Bcl-xL
- 125 Downloads
Fluorouracil (5-FU) which has been widely used in postoperative adjuvant therapy in patients with colon cancer, remains the main backbone of combination treatment of patients with colon cancer. However, the efficacy of 5-FU alone in colorectal cancer patients with BRAFV600E is not clear. In this study, we demonstrated that BRAFV600E confers sensitivity to 5-FU in vitro and in vivo xenograft model, using the paired isogenic colorectal cancer cell lines RKO with either BRAF Wild Type (WT)(+/−) or mutant (Mut) (600E/–). Our results revealed 5-FU preferably induces marked apoptosis in BRAF-mutant colorectal cancer cells, through attenuating expression of Bcl-xL and activation caspase-3/9 pathway, eventually conferring the anti-tumor efficacy of 5-FU in vitro and in vivo. Meanwhile, expression of Bcl-xL remained unchanged in BRAF WT group after treatment of 5-FU, although low extent of anti-tumor activity of 5-FU still being observed. In conclusion, these results provided a better understanding of clinical outcome of 5-FU between BRAF WT and mutant colorectal cancer patients, and suggested the inhibition of Bcl-xL might present an alternative strategy to enhance the therapeutic efficacy of 5-FU in colorectal cancer patients with BRAF mutation.
KeywordsColorectal cancer BRAF V600E 5-FU Apoptosis Bcl-xL
Mitogen-activated protein kinase
We thank Bert Vogelstein (Howard Hughes Medical Institute, Johns Hopkins University) for Isogenic RKO BRAF WT and V600E mutant cells. This work is supported by National Natural Science Foundation of China (81603136) and Jilin Provincial Commission of Health and Family Planning (2016J068).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
All animal care and experiments were conducted in accordance with national and institutional policies for animal health and well-being. The experimental procedure was performed by the Animal Ethics Committee of College of Basic Medical Sciences at Jilin University.
- 3.Rizzo S, Bronte G, Fanale D, Corsini L, Silvestris N, Santini D, Gulotta G, Bazan V, Gebbia N, Fulfaro F, Russo A (2010) Prognostic vs predictive molecular biomarkers in colorectal cancer: is KRAS and BRAF wild type status required for anti-EGFR therapy? Cancer Treat Rev 36(Suppl 3):S56–S61. https://doi.org/10.1016/s0305-7372(10)70021-9 CrossRefPubMedGoogle Scholar
- 4.Tejpar S, Bertagnolli M, Bosman F, Lenz HJ, Garraway L, Waldman F, Warren R, Bild A, Collins-Brennan D, Hahn H, Harkin DP, Kennedy R, Ilyas M, Morreau H, Proutski V, Swanton C, Tomlinson I, Delorenzi M, Fiocca R, Van Cutsem E, Roth A (2010) Prognostic and predictive biomarkers in resected colon cancer: current status and future perspectives for integrating genomics into biomarker discovery. Oncologist 15(4):390–404. https://doi.org/10.1634/theoncologist.2009-0233 CrossRefPubMedPubMedCentralGoogle Scholar
- 5.Bollag G, Hirth P, Tsai J, Zhang J, Ibrahim PN, Cho H, Spevak W, Zhang C, Zhang Y, Habets G, Burton EA, Wong B, Tsang G, West BL, Powell B, Shellooe R, Marimuthu A, Nguyen H, Zhang KY, Artis DR, Schlessinger J, Su F, Higgins B, Iyer R, D’Andrea K, Koehler A, Stumm M, Lin PS, Lee RJ, Grippo J, Puzanov I, Kim KB, Ribas A, McArthur GA, Sosman JA, Chapman PB, Flaherty KT, Xu X, Nathanson KL, Nolop K (2010) Clinical efficacy of a RAF inhibitor needs broad target blockade in BRAF-mutant melanoma. Nature 467(7315):596–599. https://doi.org/10.1038/nature09454 CrossRefPubMedPubMedCentralGoogle Scholar
- 6.Sosman JA, Kim KB, Schuchter L, Gonzalez R, Pavlick AC, Weber JS, McArthur GA, Hutson TE, Moschos SJ, Flaherty KT, Hersey P, Kefford R, Lawrence D, Puzanov I, Lewis KD, Amaravadi RK, Chmielowski B, Lawrence HJ, Shyr Y, Ye F, Li J, Nolop KB, Lee RJ, Joe AK, Ribas A (2012) Survival in BRAF V600-mutant advanced melanoma treated with vemurafenib. N Engl J Med 366(8):707–714. https://doi.org/10.1056/NEJMoa1112302 CrossRefPubMedPubMedCentralGoogle Scholar
- 7.Prahallad A, Sun C, Huang S, Di Nicolantonio F, Salazar R, Zecchin D, Beijersbergen RL, Bardelli A, Bernards R (2012) Unresponsiveness of colon cancer to BRAF(V600E) inhibition through feedback activation of EGFR. Nature 483(7387):100–103. https://doi.org/10.1038/nature10868 CrossRefPubMedGoogle Scholar
- 9.Noordhuis P, Holwerda U, Van der Wilt CL, Van Groeningen CJ, Smid K, Meijer S, Pinedo HM, Peters GJ (2004) 5-Fluorouracil incorporation into RNA and DNA in relation to thymidylate synthase inhibition of human colorectal cancers. Ann Oncol 15(7):1025–1032. https://doi.org/10.1093/annonc/mdh264 CrossRefPubMedGoogle Scholar
- 13.Sun C, Wang L, Huang S, Heynen GJ, Prahallad A, Robert C, Haanen J, Blank C, Wesseling J, Willems SM, Zecchin D, Hobor S, Bajpe PK, Lieftink C, Mateus C, Vagner S, Grernrum W, Hofland I, Schlicker A, Wessels LF, Beijersbergen RL, Bardelli A, Di Nicolantonio F, Eggermont AM, Bernards R (2014) Reversible and adaptive resistance to BRAF(V600E) inhibition in melanoma. Nature 508(7494):118–122. https://doi.org/10.1038/nature13121 CrossRefPubMedGoogle Scholar
- 18.Corcoran RB, Ebi H, Turke AB, Coffee EM, Nishino M, Cogdill AP, Brown RD, Della Pelle P, Dias-Santagata D, Hung KE, Flaherty KT, Piris A, Wargo JA, Settleman J, Mino-Kenudson M, Engelman JA (2012) EGFR-mediated re-activation of MAPK signaling contributes to insensitivity of BRAF mutant colorectal cancers to RAF inhibition with vemurafenib. Cancer Discov 2(3):227–235. https://doi.org/10.1158/2159-8290.cd-11-0341 CrossRefPubMedPubMedCentralGoogle Scholar
- 20.Koukourakis MI, Giatromanolaki A, Sivridis E, Pitiakoudis M, Gatter KC, Harris AL (2010) Beclin 1 over- and underexpression in colorectal cancer: distinct patterns relate to prognosis and tumour hypoxia. Br J Cancer 103(8):1209–1214. https://doi.org/10.1038/sj.bjc.6605904 CrossRefPubMedPubMedCentralGoogle Scholar
- 21.De Simone V, Franze E, Ronchetti G, Colantoni A, Fantini MC, Di Fusco D, Sica GS, Sileri P, MacDonald TT, Pallone F, Monteleone G, Stolfi C (2015) Th17-type cytokines, IL-6 and TNF-alpha synergistically activate STAT3 and NF-kB to promote colorectal cancer cell growth. Oncogene 34(27):3493–3503. https://doi.org/10.1038/onc.2014.286 CrossRefPubMedGoogle Scholar
- 26.Zhang H, Xue J, Hessler P, Tahir SK, Chen J, Jin S, Souers AJ, Leverson JD, Lam LT (2015) Genomic analysis and selective small molecule inhibition identifies BCL-X(L) as a critical survival factor in a subset of colorectal cancer. Mol Cancer 14:126. https://doi.org/10.1186/s12943-015-0397-y CrossRefPubMedPubMedCentralGoogle Scholar
- 27.Uppada SB, Gowrikumar S, Ahmad R, Kumar B, Szeglin B, Chen X, Smith JJ, Batra SK, Singh AB, Dhawan P (2018) MASTL induces colon cancer progression and chemoresistance by promoting Wnt/beta-catenin signaling. Mol Cancer 17(1):111. https://doi.org/10.1186/s12943-018-0848-3 CrossRefPubMedPubMedCentralGoogle Scholar
- 28.Vesel M, Rapp J, Feller D, Kiss E, Jaromi L, Meggyes M, Miskei G, Duga B, Smuk G, Laszlo T, Karner I, Pongracz JE (2017) ABCB1 and ABCG2 drug transporters are differentially expressed in non-small cell lung cancers (NSCLC) and expression is modified by cisplatin treatment via altered Wnt signaling. Respir Res 18(1):52. https://doi.org/10.1186/s12931-017-0537-6 CrossRefPubMedPubMedCentralGoogle Scholar