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Open Access 04-11-2022 | Melanoma | Original Article

Silencing FOXP2 reverses vemurafenib resistance in BRAF^V600E mutant papillary thyroid cancer and melanoma cells

Authors: Suyuan Jiang, Yuxin Huang, Yuan Li, Qin Gu, Cuiping Jiang, Xiaoming Tao, Jiao Sun

Published in: Endocrine | Issue 1/2023

Abstract

Background

Vemurafenib (VEM) is a commonly used inhibitor of papillary thyroid cancer (PTC) and melanoma with the BRAF^V600E mutation; however, acquired resistance is unavoidable. The present study aimed to identify a potential target to reverse resistance.

Materials and methods

A VEM-resistant PTC cell line (B-CPAP/VR) was established by gradually increasing the drug concentration, and a VEM-resistant BRAF^V600E melanoma cell line (A375/VR) was also established. RNA sequencing and bioinformatics analyses were conducted to identify dysregulated genes and construct a transcription factor (TF) network. The role of a potential TF, forkhead box P2 (FOXP2), verified by qRT-PCR, was selected for further confirmation.

Results

The two resistant cell lines were tolerant of VEM and displayed higher migration and colony formation abilities (p < 0.05). RNA sequencing identified 9177 dysregulated genes in the resistant cell lines, and a TF network consisting of 13 TFs and 44 target genes was constructed. Alterations in FOXP2 expression were determined to be consistent between the two VEM-resistant cell lines. Finally, silencing FOXP2 resulted in an increase in drug sensitivity and significant suppression of the migration and colony formation abilities of the two resistant cell lines (p < 0.05).

Conclusions

The present study successfully established two VEM-resistant cell lines and identified a potential target for VEM-resistant PTC or melanoma.

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J. Hu, I.J. Yuan, S. Mirshahidi, A. Simental, S.C. Lee, X. Yuan. Thyroid Carcinoma: Phenotypic Features, Underlying Biology and Potential Relevance for Targeting Therapy. Int. J. Mol. Sci. 22(4), (2021). https://doi.org/10.3390/ijms22041950

C. Romei, R. Ciampi, R. Elisei, A comprehensive overview of the role of the RET proto-oncogene in thyroid carcinoma. Nat. Rev. Endocrinol. 12(4), 192–202 (2016). https://doi.org/10.1038/nrendo.2016.11CrossRef

M.I. Abdullah, S.M. Junit, K.L. Ng, J.J. Jayapalan, B. Karikalan, O.H. Hashim, Papillary Thyroid Cancer: Genetic alterations and molecular biomarker investigations. Int J. Med Sci. 16(3), 450–460 (2019). https://doi.org/10.7150/ijms.29935CrossRef

E.T. Kimura, M.N. Nikiforova, Z. Zhu, J.A. Knauf, Y.E. Nikiforov, J.A. Fagin, High prevalence of BRAF mutations in thyroid cancer: genetic evidence for constitutive activation of the RET/PTC-RAS-BRAF signaling pathway in papillary thyroid carcinoma. Cancer Res. 63(7), 1454–1457 (2003)

R.H. Grogan, E.J. Mitmaker, O.H. Clark, The evolution of biomarkers in thyroid cancer-from mass screening to a personalized biosignature. Cancers (Basel) 2(2), 885–912 (2010). https://doi.org/10.3390/cancers2020885CrossRef

Y.E. Nikiforov, M.N. Nikiforova, Molecular genetics and diagnosis of thyroid cancer. Nat. Rev. Endocrinol. 7(10), 569–580 (2011). https://doi.org/10.1038/nrendo.2011.142CrossRef

S. Fraser, C. Go, A. Aniss, S. Sidhu, L. Delbridge, D. Learoyd et al. BRAF(V600E) mutation is associated with decreased disease-free survival in papillary thyroid cancer. World J. Surg. 40(7), 1618–1624 (2016). https://doi.org/10.1007/s00268-016-3534-xCrossRef

M. Xing, A.S. Alzahrani, K.A. Carson, Y.K. Shong, T.Y. Kim, D. Viola et al. Association between BRAF V600E mutation and recurrence of papillary thyroid cancer. J. Clin. Oncol. 33(1), 42–50 (2015). https://doi.org/10.1200/JCO.2014.56.8253CrossRef

C. Garbe, T.K. Eigentler, Vemurafenib. Recent Results Cancer Res. 211, 77–89 (2018). https://doi.org/10.1007/978-3-319-91442-8_6CrossRef

10.

K.B. Kim, M.E. Cabanillas, A.J. Lazar, M.D. Williams, D.L. Sanders, J.L. Ilagan et al. Clinical responses to vemurafenib in patients with metastatic papillary. Thyroid cancer harboring BRAF(V600E) Mutat. Thyroid 23(10), 1277–1283 (2013). https://doi.org/10.1089/thy.2013.0057CrossRef

11.

G. Bollag, P. Hirth, J. Tsai, J. Zhang, P.N. Ibrahim, H. Cho et al. Clinical efficacy of a RAF inhibitor needs broad target blockade in BRAF-mutant melanoma. Nature 467(7315), 596–599 (2010). https://doi.org/10.1038/nature09454CrossRef

12.

G. Bollag, J. Tsai, J. Zhang, C. Zhang, P. Ibrahim, K. Nolop et al. Vemurafenib: the first drug approved for BRAF-mutant cancer. Nat. Rev. Drug Disco. 11(11), 873–886 (2012). https://doi.org/10.1038/nrd3847CrossRef

13.

M.S. Brose, M.E. Cabanillas, E.E. Cohen, L.J. Wirth, T. Riehl, H. Yue et al. Vemurafenib in patients with BRAF(V600E)-positive metastatic or unresectable papillary thyroid cancer refractory to radioactive iodine: a non-randomised, multicentre, open-label, phase 2 trial. Lancet Oncol. 17(9), 1272–1282 (2016). https://doi.org/10.1016/S1470-2045(16)30166-8CrossRef

14.

J.J. Harding, F. Catalanotti, R.R. Munhoz, D.T. Cheng, A. Yaqubie, N. Kelly et al. A retrospective evaluation of vemurafenib as treatment for BRAF-mutant melanoma brain metastases. Oncologist 20(7), 789–797 (2015). https://doi.org/10.1634/theoncologist.2014-0012CrossRef

15.

F. Crispo, T. Notarangelo, M. Pietrafesa, G. Lettini, G. Storto, A. Sgambato, et al. BRAF inhibitors in thyroid cancer: Clinical impact, mechanisms of resistance and future perspectives. Cancers (Basel) 11(9), (2019). https://doi.org/10.3390/cancers11091388

16.

M. Duquette, P.M. Sadow, A. Husain, J.N. Sims, Z.A. Antonello, A.H. Fischer et al. Metastasis-associated MCL1 and P16 copy number alterations dictate resistance to vemurafenib in a BRAFV600E patient-derived papillary thyroid carcinoma preclinical model. Oncotarget 6(40), 42445–42467 (2015). https://doi.org/10.18632/oncotarget.6442CrossRef

17.

M.A. Roelli, D. Ruffieux-Daidie, A. Stooss, O. ElMokh, W.A. Phillips, M.S. Dettmer et al. PIK3CA(H1047R)-induced paradoxical ERK activation results in resistance to BRAF(V600E) specific inhibitors in BRAF(V600E) PIK3CA(H1047R) double mutant thyroid tumors. Oncotarget 8(61), 103207–103222 (2017). https://doi.org/10.18632/oncotarget.21732CrossRef

18.

J.H. Kim, J. Hwang, J.H. Jung, H.J. Lee, D.Y. Lee, S.H. Kim, Molecular networks of FOXP family: Dual biologic functions, interplay with other molecules and clinical implications in cancer progression. Mol. Cancer 18(1), 180 (2019). https://doi.org/10.1186/s12943-019-1110-3CrossRef

19.

Y. Liu, T. Chen, M. Guo, Y. Li, Q. Zhang, G. Tan et al. FOXA2-Interacting FOXP2 prevents epithelial-mesenchymal transition of breast cancer cells by stimulating E-Cadherin and PHF2 Transcription. Front Oncol. 11, 605025 (2021). https://doi.org/10.3389/fonc.2021.605025CrossRef

20.

J. Wu, P. Liu, H. Tang, Z. Shuang, Q. Qiu, L. Zhang et al. FOXP2 promotes tumor proliferation and metastasis by targeting GRP78 in triple-negative breast cancer. Curr. Cancer Drug Targets 18(4), 382–389 (2018). https://doi.org/10.2174/1568009618666180131115356CrossRef

21.

W. Chang, Q. Chang, H. Lu, Y. Li, C. Chen, MiR-221-3p facilitates thyroid cancer cell proliferation and inhibit apoptosis by targeting FOXP2 through hedgehog pathway. Mol. Biotechnol. 64(8), 919–927 (2022). https://doi.org/10.1007/s12033-022-00473-5CrossRef

22.

J. Zhou, L. Zhang, H. Zheng, W. Ge, Y. Huang, Y. Yan et al. Identification of chemoresistance-related mRNAs based on gemcitabine-resistant pancreatic cancer cell lines. Cancer Med. 9(3), 1115–1130 (2020). https://doi.org/10.1002/cam4.2764CrossRef

23.

Y. Koh, W.J. Jung, K.S. Ahn, S.S. Yoon, Establishment of cell lines from both myeloma bone marrow and plasmacytoma: SNU_MM1393_BM and SNU_MM1393_SC from a single patient. Biomed. Res. Int. 2014, 510408 (2014). https://doi.org/10.1155/2014/510408CrossRef

24.

S.A. Luebker, S.A. Koepsell, Diverse mechanisms of BRAF inhibitor resistance in melanoma identified in clinical and preclinical studies. Front. Oncol. 9, 268 (2019). https://doi.org/10.3389/fonc.2019.00268CrossRef

25.

H. Luo, M. Umebayashi, K. Doi, T. Morisaki, S. Shirasawa, T. Tsunoda,, Resveratrol Overcomes Cellular Resistance to Vemurafenib Through Dephosphorylation of AKT in BRAF-mutated Melanoma Cells. Anticancer Res 36(7), 3585–3589 (2016).

26.

N. Wang, J. Wen, W. Ren, Y. Wu, C. Deng. Upregulation of TRIB2 by Wnt/beta-catenin activation in BRAF(V600E) papillary thyroid carcinoma cells confers resistance to BRAF inhibitor vemurafenib. Cancer Chemother. Pharm. 88(1), 155–164 (2021). https://doi.org/10.1007/s00280-021-04270-wCrossRef

27.

M. Zerfaoui, T.M. Dokunmu, E.A. Toraih, B.M. Rezk, Z.Y. Abd Elmageed, E. Kandil. New Insights into the Link between Melanoma and Thyroid Cancer: Role of Nucleocytoplasmic Trafficking. Cells 10(2), (2021). https://doi.org/10.3390/cells10020367

28.

A. Porter, D.J. Wong, Perspectives on the treatment of advanced thyroid cancer: Approved therapies, resistance mechanisms, and future directions. Front. Oncol. 10, 592202 (2020). https://doi.org/10.3389/fonc.2020.592202CrossRef

29.

D.S. Latchman, Transcription factors: An overview. Int J. Biochem Cell Biol. 29(12), 1305–1312 (1997). https://doi.org/10.1016/s1357-2725(97)00085-xCrossRef

30.

F. Yang, Z. Xiao, S. Zhang, FOXP2 regulates thyroid cancer cell proliferation and apoptosis via transcriptional activation of RPS6KA6. Exp. Ther. Med 23(6), 434 (2022). https://doi.org/10.3892/etm.2022.11361CrossRef

31.

R. Huang, G. Xiang, X. Duan, H. Wang, K. He, J. Xiao, MiR-132-3p inhibits proliferation, invasion and migration of colorectal cancer cells via down-regulating FOXP2 expression. Acta Biochim Pol. 69(2), 371–377 (2022). https://doi.org/10.18388/abp.2020_5813CrossRef

32.

K.K. Wong, D.M. Gascoyne, E.J. Soilleux, L. Lyne, H. Spearman, G. Roncador et al. FOXP2-positive diffuse large B-cell lymphomas exhibit a poor response to R-CHOP therapy and distinct biological signatures. Oncotarget 7(33), 52940–52956 (2016). https://doi.org/10.18632/oncotarget.9507CrossRef

33.

X.L. Song, Y. Tang, X.H. Lei, S.C. Zhao, Z.Q. Wu, miR-618 Inhibits Prostate Cancer Migration and Invasion by Targeting FOXP2. J. Cancer 8(13), 2501–2510 (2017). https://doi.org/10.7150/jca.17407CrossRef

34.

W.Z. Jia, T. Yu, Q. An, H. Yang, Z. Zhang, X. Liu et al. MicroRNA-190 regulates FOXP2 genes in human gastric cancer. Onco Targets Ther. 9, 3643–3651 (2016). https://doi.org/10.2147/OTT.S103682CrossRef

35.

K. Hama, K. Bandoh, Y. Kakehi, J. Aoki, H. Arai, Lysophosphatidic acid (LPA) receptors are activated differentially by biological fluids: possible role of LPA-binding proteins in activation of LPA receptors. FEBS Lett. 523(1–3), 187–192 (2002). https://doi.org/10.1016/s0014-5793(02)02976-9CrossRef

Title: Silencing FOXP2 reverses vemurafenib resistance in BRAFV600E mutant papillary thyroid cancer and melanoma cells
Authors: Suyuan Jiang
Yuxin Huang
Yuan Li
Qin Gu
Cuiping Jiang
Xiaoming Tao
Jiao Sun
Publication date: 04-11-2022
Publisher: Springer US
Keywords: Melanoma
Melanoma
Thyroid Cancer
Thyroid Cancer
Thyroid Cancer
Published in: Endocrine / Issue 1/2023
Print ISSN: 1355-008X
Electronic ISSN: 1559-0100
DOI: https://doi.org/10.1007/s12020-022-03180-y

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Silencing FOXP2 reverses vemurafenib resistance in BRAF^V600E mutant papillary thyroid cancer and melanoma cells

Abstract

Background

Materials and methods

Results

Conclusions

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Abstract

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Conclusions

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