<em>In vitro</em> anticancer effects of alpelisib against PIK3CA‑mutated canine hemangiosarcoma cell lines

Maeda,Marika; Maeda,Marika; Ochiai,Kazuhiko; Ochiai,Kazuhiko; Michishita,Masaki; Michishita,Masaki; Morimatsu,Masami; Morimatsu,Masami; Sakai,Hiroki; Sakai,Hiroki; Kinoshita,Nayuta; Kinoshita,Nayuta; Sakaue,Motoharu; Sakaue,Motoharu; Onozawa,Eri; Onozawa,Eri; Azakami,Daigo; Azakami,Daigo; Yamamoto,Masami; Yamamoto,Masami; Ishioka,Katsumi; Ishioka,Katsumi; Sadahira,Takuya; Sadahira,Takuya; Watanabe,Masami; Watanabe,Masami; Tanaka,Yoshikazu; Tanaka,Yoshikazu

doi:10.3892/or.2022.8295

In vitro anticancer effects of alpelisib against PIK3CA‑mutated canine hemangiosarcoma cell lines

Authors:
- Marika Maeda
- Kazuhiko Ochiai
- Masaki Michishita
- Masami Morimatsu
- Hiroki Sakai
- Nayuta Kinoshita
- Motoharu Sakaue
- Eri Onozawa
- Daigo Azakami
- Masami Yamamoto
- Katsumi Ishioka
- Takuya Sadahira
- Masami Watanabe
- Yoshikazu Tanaka
View Affiliations

Affiliations: Laboratory of Veterinary Hygiene, School of Veterinary Science, Nippon Veterinary and Life Science University, Tokyo 180‑8602, Japan, Research Center for Animal Life Science, Nippon Veterinary and Life Science University, Tokyo 180‑8602, Japan, Laboratory of Laboratory Animal Science and Medicine, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Hokkaido 060‑0818, Japan, Laboratory of Veterinary Pathology, Gifu University, Gifu 501‑1193, Japan, Laboratory of Anatomy II, Department of Veterinary Medicine, Azabu University, Sagamihara-shi, Kanagawa 252‑5201, Japan, School of Veterinary Nursing and Technology, Nippon Veterinary and Life Science University, Tokyo 180‑8602, Japan, Laboratory of Clinical Oncology, Tokyo University of Agriculture and Technology, Tokyo 183‑8538, Japan, Division of Physiological Pathology, Nippon Veterinary and Life Science University, Tokyo 180‑8602, Japan, Department of Urology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama 700‑0914, Japan
Published online on: February 25, 2022 https://doi.org/10.3892/or.2022.8295
Article Number: 84
Copyright: © Maeda 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

Hemangiosarcoma (HSA) is a malignant neoplasm that occurs in humans and canines with a poor prognosis owing to metastatic spread, despite effective treatment. The frequency of spontaneous HSA development is higher in canines than in humans. Therefore, canine HSA is a useful model of intractable human disease, which requires early detection and an effective therapeutic strategy. A high frequency of the p110α phosphatidylinositol‑4,5‑bisphosphate 3‑kinase catalytic subunit alpha (PIK3CA) mutations is detected in a comprehensive genome‑wide analysis of canine cases of HSA. The present cloned the full‑length cDNA of canine PIK3CA and identified a mutation in codon 1047 from canine cases of HSA and cell lines that were established from these. The enforced expression of the 1047th histidine residue (H1047)R or L mutants of canine PIK3CA in HeLa cells enhanced epidermal growth factor receptor (EGFR) signaling via Akt phosphorylation. PIK3CA mutant canine HSA cell lines exhibited the hyperphosphorylation of Akt upon EGF stimulation as well. Alpelisib, a molecular targeted drug against PIK3CA activating mutations, exerted a significant antitumor effect in canine PIK3CA‑mutated HSA cell lines. By contrast, it had no significant effect on canine mammary gland tumor cell lines harboring PIK3CA mutations. On the whole, the findings of the present study suggest that alpelisib may be highly effective against PIK3CA mutations that occur frequently in canine HSA.

View Figures

View References

1	Itakura E, Yamamoto H, Oda Y and Tsuneyoshi M: Detection and characterization of vascular endothelial growth factors and their receptors in a series of angiosarcomas. J Surg Oncol. 97:74–81. 2008. View Article : Google Scholar : PubMed/NCBI
2	Clifford CA, Mackin AJ and Henry CJ: Treatment of canine hemangiosarcoma: 2000 and beyond. J Vet Intern Med. 14:479–485. 2000. View Article : Google Scholar : PubMed/NCBI
3	Yonemaru K, Sakai H, Murakami M, Yanai T and Masegi T: Expression of vascular endothelial growth factor, basic fibroblast growth factor, and their receptors (flt-1, flk-1, and flg-1) in canine vascular tumors. Vet Pathol. 43:971–980. 2006. View Article : Google Scholar : PubMed/NCBI
4	Chappell WH, Steelman LS, Long JM, Kempf RC, Abrams SL, Franklin RA, Bäsecke J, Stivala F, Donia F, Fagone P, et al: Ras/Raf/MEK/ERK and PI3K/PTEN/Akt/mTOR inhibitors: Rationale and importance to inhibiting these pathways in human health. Oncotarget. 2:135–164. 2011. View Article : Google Scholar : PubMed/NCBI
5	di Blasio L, Puliafito A, Gagliardi PA, Comunanza V, Somale D, Chiaverina G, Bussolino F and Primo L: PI3K/mTOR inhibition promotes the regression of experimental vascular malformations driven by PIK3CA-activating mutations. Cell Death Dis. 9:452018. View Article : Google Scholar : PubMed/NCBI
6	Matsumura I, Mizuki M and Kanakura Y: Roles for deregulated receptor tyrosine kinases and their downstream signaling molecules in hematologic malignancies. Cancer Sci. 99:479–485. 2008. View Article : Google Scholar : PubMed/NCBI
7	Arbiser JL, Weiss SW, Arbiser ZK, Bravo F, Govindajaran B, Caceres-Rios H, Cotsonis G, Recavarren S, Swerlick RA and Cohen C: Differential expression of active mitogen-activated protein kinase in cutaneous endothelial neoplasms: Implications for biologic behavior and response to therapy. J Am Acad Dermatol. 44:193–197. 2001. View Article : Google Scholar : PubMed/NCBI
8	Kroll J and Waltenberger J: The vascular endothelial growth factor receptor KDR activates multiple signal transduction pathways in porcine aortic endothelial cells. J Biol Chem. 272:32521–32527. 1997. View Article : Google Scholar : PubMed/NCBI
9	Jiang X, Wang J, Deng X, Xiong F, Zhang S, Gong Z, Li X, Cao K, Deng H, He Y, et al: The role of microenvironment in tumor angiogenesis. J Exp Clin Cancer Res. 39:2042020. View Article : Google Scholar : PubMed/NCBI
10	Blasio A, Wang J, Wang D, Varodayan FP, Pomrenze MB, Miller J, Lee AM, McMahon T, Gyawali S, Wang HY, et al: Novel small-molecule inhibitors of protein kinase C epsilon reduce ethanol consumption in mice. Biol Psychiatry. 84:193–201. 2018. View Article : Google Scholar : PubMed/NCBI
11	Le Cras TD, Goines J, Lakes N, Pastura P, Hammill AM, Adams DM and Boscolo E: Constitutively active PIK3CA mutations are expressed by lymphatic and vascular endothelial cells in capillary lymphatic venous malformation. Angiogenesis. 23:425–442. 2020. View Article : Google Scholar : PubMed/NCBI
12	Dickerson EB, Thomas R, Fosmire SP, Lamerato-Kozicki AR, Bianco SR, Wojcieszyn JW, Breen M, Helfand SC and Modiano JF: Mutations of phosphatase and tensin homolog deleted from chromosome 10 in canine hemangiosarcoma. Vet Pathol. 42:618–632. 2005. View Article : Google Scholar : PubMed/NCBI
13	Tate G, Suzuki T and Mitsuya T: Mutation of the PTEN gene in a human hepatic angiosarcoma. Cancer Genet Cytogenet. 178:160–162. 2007. View Article : Google Scholar : PubMed/NCBI
14	Lahat G, Dhuka AR, Hallevi H, Xiao L, Zou C, Smith KD, Phung TL, Pollock RE, Benjamin R, Hunt KK, et al: Angiosarcoma: Clinical and molecular insights. Ann Surg. 251:1098–1106. 2010. View Article : Google Scholar : PubMed/NCBI
15	Murai A, Asa SA, Kodama A, Hirata A, Yanai T and Sakai H: Constitutive phosphorylation of the mTORC2/Akt/4E-BP1 pathway in newly derived canine hemangiosarcoma cell lines. BMC Vet Res. 8:1282012. View Article : Google Scholar : PubMed/NCBI
16	Tamburini BA, Trapp S, Phang TL, Schappa JT, Hunter LE and Modiano JF: Gene expression profiles of sporadic canine hemangiosarcoma are uniquely associated with breed. PLoS One. 4:e55492009. View Article : Google Scholar : PubMed/NCBI
17	Lequarre AS, Andersson L, Andre C, Fredholm M, Hitte C, Leeb T, Lohi H, Lindblad-Toh K and Georges M: LUPA: A European initiative taking advantage of the canine genome architecture for unravelling complex disorders in both human and dogs. Vet J. 189:155–159. 2011. View Article : Google Scholar : PubMed/NCBI
18	Espejo-Freire AP, Elliott A, Rosenberg A, Costa PA, Barreto-Coelho P, Jonczak E, D'amato G, Subhawong T, Arshad J, Diaz-Perez JA, et al: Genomic landscape of angiosarcoma: A targeted and immunotherapy biomarker analysis. Cancers. 13:48162021. View Article : Google Scholar : PubMed/NCBI
19	Wang G, Wu M, Maloneyhuss MA, Wojcik J, Durham AC, Masson NJ and Royh DB: Actionable mutations in canine hemangiosarcoma. PLoS One. 12:e01886672017. View Article : Google Scholar : PubMed/NCBI
20	Arafeh R and Samuels Y: PIK3CA in cancer: The past 30 years. Semin Cancer Biol. 59:36–49. 2019. View Article : Google Scholar : PubMed/NCBI
21	André F, Ciruelos E, Rubovszky G, Campone M, Loibl S, Rugo HS, Iwata H, Conte P, Mayer IA, Kaufmanet K, et al: Alpelisib for PIK3CA-mutated, hormone receptor-positive advanced breast cancer. N Engl J Med. 380:1929–1940. 2019. View Article : Google Scholar : PubMed/NCBI
22	Misdorp W; Armed Forces Institute of Pathology (U.S.); American Registry of Pathology; WHO Collaborating Center for Worldwide Reference on Comparative Oncology, : Histological classification of mammary tumors of the dog and the cat. International Histological Classification of Tumors of Domestic Animals. Vol 7. 2nd series. Armed Forces Institute of Pathology in cooperation with the American Registry of Pathology and the World Health Organization Collaborating Center for Worldwide Reference on Comparative Oncology; Washington, DC: 1999
23	Uyama R, Nakagawa T, Hong SH, Mochizuki M, Nishimura R and Sasaki N: Establishment of four pairs of canine mammary tumour cell lines derived from primary and metastatic origin and their E-cadherin expression. Vet Comp Oncol. 4:104–113. 2006. View Article : Google Scholar : PubMed/NCBI
24	Ghazanchaei A, Mansoori B, Mohammadi A, Biglari A and Baradaran B: Restoration of miR-152 expression suppresses cell proliferation, survival, and migration through inhibition of AKT-ERK pathway in colorectal cancer. J Cell Physiol. 234:769–776. 2019. View Article : Google Scholar : PubMed/NCBI
25	Madsen RR, Vanhaesebroeck B and Semple RK: Cancer-associated PIK3CA mutations in overgrowth disorders. Trends Mol Med. 24:856–870. 2018. View Article : Google Scholar : PubMed/NCBI
26	Kim JH: PIK3CA mutations matter for cancer in dogs. Res Vet Sci. 133:39–41. 2020. View Article : Google Scholar : PubMed/NCBI
27	Ney JT, Froehner S, Roesler A, Buettner R and Merkelbach-Bruse S: High-resolution melting analysis as a sensitive prescreening diagnostic tool to detect KRAS, BRAF, PIK3CA, and AKT1 mutations in formalin-fixed, paraffin-embedded tissues. Arch Pathol Lab Med. 136:983–992. 2012. View Article : Google Scholar : PubMed/NCBI
28	Beca F, Krings G, Chen YY, Hosfield EM, Vohra P, Sibley RK, Troxell ML, West RB, Allison KH and Bean GR: Primary mammary angiosarcomas harbor frequent mutations in KDR and PIK3CA and show evidence of distinct pathogenesis. Mod Pathol. 33:1518–1526. 2020. View Article : Google Scholar : PubMed/NCBI
29	Wang G, Wu M, Durham AC, Mason NJ and Roth DB: Canine oncopanel: A capture-based, NGS platform for evaluating the mutational landscape and detecting putative driver mutations in canine cancers. Vet Comp Oncol. 20:91–101. 2022. View Article : Google Scholar : PubMed/NCBI
30	Wang J, Zhao W, Guo H, Fang Y, Stockman SE, Bai S, Ng PK, Li Y, Yu Q, Lu Y, et al: AKT isoform-specific expression and activation across cancer lineages. BMC Cancer. 18:7422018. View Article : Google Scholar : PubMed/NCBI
31	Guo H, Gao M, Lu Y, Liang J, Lorenzi PL, Bai S, Hawke DH, Li J, Dogruluk T, Scott KL, et al: Coordinate phosphorylation of multiple residues on single AKT1 and AKT2 molecules. Oncogene. 33:3463–3472. 2014. View Article : Google Scholar : PubMed/NCBI
32	Dillon RL, Marcotte R, Hennessy BT, Woodgett JR, Mills GB and Muller WJ: Akt1 and akt2 play distinct roles in the initiation and metastatic phases of mammary tumor progression. Cancer Res. 69:5057–5064. 2009. View Article : Google Scholar : PubMed/NCBI
33	Ihle MA, Fassunke J, König K, Grunewald I, Schlaak M, Kreuzberg N, Tietze L, Schildhaus HU, Büttner R and Merkelbach-Bruseet S: Comparison of high resolution melting analysis, pyrosequencing, next generation sequencing and immunohistochemistry to conventional Sanger sequencing for the detection of p.V600E and non-p.V600E BRAF mutations. BMC Cancer. 14:132014. View Article : Google Scholar : PubMed/NCBI
34	Rascio F, Spadaccino F, Rocchetti MT, Castellano G, Stallone G, Netti GS and Ranieri E: The pathogenic role of PI3K/AKT pathway in cancer onset and drug resistance: An updated review. Cancers. 13:39492021. View Article : Google Scholar : PubMed/NCBI
35	Fritsch C, Huang A, Chatenay-Rivauday C, Schnell C, Reddy A, Liu M, Kauffmann A, Guthy D, Erdmann D, De Pover A, et al: Characterization of the novel and specific PI3Kα inhibitor NVP-BYL719 and development of the patient stratification strategy for clinical trials. Mol Cancer Ther. 13:1117–1129. 2014. View Article : Google Scholar : PubMed/NCBI
36	Im KS, Graef AJ, Breen M, Lindblad-Toh K, Modiano JF and Kim JH: Interactions between CXCR4 and CXCL12 promote cell migration and invasion of canine hemangiosarcoma. Vet Comp Oncol. 15:315–327. 2017. View Article : Google Scholar : PubMed/NCBI
37	Kim JH, Graef AJ, Dickerson EB and Modiano JF: Pathobiology of hemangiosarcoma in dogs: Research advances and future perspectives. Vet Sci. 2:388–405. 2015. View Article : Google Scholar : PubMed/NCBI
38	Kim KH, Chung WS, Kim Y, Kim KS, Lee IS, Park JY, Jeong HS, Na YC, Lee CH and Jang HJ: Transcriptomic analysis reveals wound healing of morus alba root extract by up-regulating keratin filament and CXCL12/CXCR4 signaling. Phytother Res. 29:1251–1258. 2015. View Article : Google Scholar : PubMed/NCBI
39	Zuo J, Wen M, Li S, Lv X, Wang L, Ai X and Lei M: Overexpression of CXCR4 promotes invasion and migration of non-small cell lung cancer via EGFR and MMP-9. Oncol Lett. 14:7513–7521. 2017.PubMed/NCBI