Lapatinib alters the malignant phenotype of osteosarcoma cells via downregulation of the activity of the HER2-PI3K/AKT-FASN axis in vitro

Long,Xin-Hua; Zhang,Guo-Mei; Peng,Ai-Fen; Luo,Qing-Feng; Zhang,Lin; Wen,Hui‑Cai; Zhou,Rong-Ping; Gao,Song; Zhou,Yang; Liu,Zhi-Li

doi:10.3892/or.2013.2825

Lapatinib alters the malignant phenotype of osteosarcoma cells via downregulation of the activity of the HER2-PI3K/AKT-FASN axis in vitro

Authors:
- Xin-Hua Long
- Guo-Mei Zhang
- Ai-Fen Peng
- Qing-Feng Luo
- Lin Zhang
- Hui‑Cai Wen
- Rong-Ping Zhou
- Song Gao
- Yang Zhou
- Zhi-Li Liu
View Affiliations

Affiliations: Department of Orthopedics, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China, Department of Orthopedics, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China, Jiangxi University of Traditional Chinese Medicine, Nanchang, Jiangxi 330006, P.R. China , Department of Pathology, Cancer Hospital of Jiangxi Province, Nanchang, Jiangxi 330006, P.R. China
Published online on: October 29, 2013 https://doi.org/10.3892/or.2013.2825
Pages: 328-334

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

Lapatinib, an inhibitor of human epidermal growth factor receptor 2 (HER2) phosphorylation, has been reported to inhibit several types of tumors such as HER2-overexpressing breast cancer. However, the effect of lapatinib on the malignant phenotype of human osteosarcoma (OS) cells and the potential molecular mechanisms remain unclear. To elucidate the effect of lapatinib on OS, two OS cell lines, U2-OS and MG-63, were utilized in the present study. Various concentrations of lapatinib were used to treat OS cells for different time durations. Cell proliferation was evaluated by MTT and colony formation assays. Flow cytometry (FCM) was used to evaluate cell apoptosis. Wound healing and Transwell invasion assays were performed to examine the migratory and invasive abilities of the cells. To investigate the possible molecular mechanisms involved, the expression of p-HER2, phosphatidylinositol 3-kinase (PI3K), p-AKT, AKT and fatty acid synthase (FASN) protein was detected by western blotting. MTT assays showed that lapatinib inhibited the proliferation of U2-OS and MG-63 cells in a dose- and time-dependent manner, and the rate of colony formation of the lapatinib-treated cells was significantly lower when compared to those cells not treated with lapatinib in both cell lines. FCM assay revealed a significantly higher apoptotic rate in the lapatinib-treated OS cells. Wound healing and Transwell invasion assays revealed that the migratory and invasive abilities of OS cells were significantly inhibited by lapatinib (P<0.05). Western blotting showed that lapatinib suppressed the activity of HER2-PI3K/AKT-FASN in U2-OS and MG-63 cells in vitro. These results suggest that lapatinib may alter the malignant phenotype of OS cells via downregulation of the activity of the HER2-PI3K/AKT-FASN signaling pathway in vitro. Thus, lapatinib may be an effective chemotherapeutic agent for the treatment of osteosarcoma.

View Figures

View References

1	Meyers PA, Schwartz CL, Krailo M, Kleinerman ES, Betcher D, Bernstein ML, et al: Osteosarcoma: a randomized, prospective trial of the addition of ifosfamide and/or muramyl tripeptide to cisplatin, doxorubicin, and high-dose methotrexate. J Clin Oncol. 23:2004–2011. 2005. View Article : Google Scholar : PubMed/NCBI
2	Bacci G, Forni C, Longhi A, Ferrari S, Mercuri M, Bertoni F, et al: Local recurrence and local control of non-metastatic osteosarcoma of the extremities: a 27-year experience in a single institution. J Surg Oncol. 96:118–123. 2007.PubMed/NCBI
3	Jawad MU, Cheung MC, Clarke J, Koniaris LG and Scully SP: Osteosarcoma: improvement in survival limited to high-grade patients only. J Cancer Res Clin Oncol. 137:597–607. 2011. View Article : Google Scholar : PubMed/NCBI
4	Brand RA: 50 years ago in CORR: the present trend in treatment of osteogenic sarcoma. Clin Orthop Relat Res. 467:3038–3039. 2009.PubMed/NCBI
5	Ferrari S, Smeland S, Mercuri M, Bertoni F, Longhi A, Ruggieri P, et al: Neoadjuvant chemotherapy with high-dose Ifosfamide, high-dose methotrexate, cisplatin, and doxorubicin for patients with localized osteosarcoma of the extremity: a joint study by the Italian and Scandinavian Sarcoma Groups. J Clin Oncol. 23:8845–8852. 2005. View Article : Google Scholar
6	Gradishar WJ: Emerging approaches for treating HER2-positive metastatic breast cancer beyond trastuzumab. Ann Oncol. 24:2492–2500. 2013. View Article : Google Scholar : PubMed/NCBI
7	Shanmughapriya S, Senthilkumar G, Arun S, Vinodhini K, Sudhakar S and Natarajaseenivasan K: Polymorphism and overexpression of HER2/neu among ovarian carcinoma women from Tiruchirapalli, Tamil Nadu, India. Arch Gynecol Obstet. May 31–2013.(Epub ahead of print).
8	Zhou BG, Liu MY, Qiu XC, Xu YM, Fan QY, Yang AG, et al: A novel recombinant immunocasp-6 fusion gene specifically and efficiently suppresses HER2-overexpressing osteosarcoma. Oncol Rep. 29:276–282. 2013.PubMed/NCBI
9	Ma Q, Zhou Y, Ma B, Chen X, Wen Y, Liu Y, et al: The clinical value of CXCR4, HER2 and CD44 in human osteosarcoma: a pilot study. Oncol Lett. 3:797–801. 2012.PubMed/NCBI
10	Scotlandi K, Manara MC, Hattinger CM, Benini S, Perdichizzi S, Pasello M, et al: Prognostic and therapeutic relevance of HER2 expression in osteosarcoma and Ewing’s sarcoma. Eur J Cancer. 41:1349–1361. 2005.PubMed/NCBI
11	Sheng WQ, Huang D, Ying JM, Lu N, Wu HM, Liu YH, et al: HER2 status in gastric cancers: a retrospective analysis from four Chinese representative clinical centers and assessment of its prognostic significance. Ann Oncol. 24:2360–1264. 2013. View Article : Google Scholar
12	Baselga J and Swain SM: Novel anticancer targets: revisiting ERBB2 and discovering ERBB3. Nat Rev Cancer. 9:463–475. 2009. View Article : Google Scholar : PubMed/NCBI
13	Morris CD, Gorlick R, Huvos G, Heller G, Meyers PA and Healey JH: Human epidermal growth factor receptor 2 as a prognostic indicator in osteogenic sarcoma. Clin Orthop Relat Res. 382:59–65. 2001. View Article : Google Scholar : PubMed/NCBI
14	Shan LQ, Ma S, Qiu XC, Wang T, Yu SB, Ma BA, et al: A novel recombinant immuno-tBid with a furin site effectively suppresses the growth of HER2-positive osteosarcoma cells in vitro. Oncol Rep. 25:325–331. 2011.PubMed/NCBI
15	Burris HA III, Hurwitz HI, Dees EC, Dowlati A, Blackwel KL, O’Neil B, et al: Phase I safety, pharmacokinetics, and clinical activity study of lapatinib (GW572016), a reversible dual inhibitor of epidermal growth factor receptor tyrosine kinases, in heavily pretreated patients with metastatic carcinomas. J Clin Oncol. 23:5305–5313. 2005. View Article : Google Scholar
16	Bence AK, Anderson EB, Halepota MA, Doukas MA, DeSimone PA, Davis GA, et al: Phase I pharmacokinetic studies evaluating single and multiple doses of oral GW572016, a dual EGFR-ErbB2 inhibitor, in healthy subjects. Invest New Drugs. 23:39–49. 2005. View Article : Google Scholar : PubMed/NCBI
17	Spector NL, Xia W, Burris H III, Hurwitz H, Dees EC, Dowlati A, et al: Study of the biologic effects of lapatinib, a reversible inhibitor of ErbB1 and ErbB2 tyrosine kinases, on tumor growth and survival pathways in patients with advanced malignancies. J Clin Oncol. 23:2502–2512. 2005. View Article : Google Scholar : PubMed/NCBI
18	Rusnak DW, Lackey K, Affleck K, Wood ER, Alligood KJ, Rhodes N, et al: The effects of the novel, reversible epidermal growth factor receptor/ErbB-2 tyrosine kinase inhibitor, GW2016, on the growth of human normal and tumor-derived cell lines in vitro and in vivo. Mol Cancer Ther. 1:85–94. 2001.PubMed/NCBI
19	Kalous O, Conklin D, Desai AJ, O’Brien NA, Ginther C, Anderson L, et al: Dacomitinib (PF-00299804), an irreversible Pan-HER inhibitor, inhibits proliferation of HER2-amplified breast cancer cell lines resistant to trastuzumab and lapatinib. Mol Cancer Ther. 11:1978–1987. 2012. View Article : Google Scholar : PubMed/NCBI
20	Ryan Q, Ibrahim A, Cohen MH, Johnson J, Ko CW, Sridhara R, et al: FDA drug approval summary: lapatinib in combination with capecitabine for previously treated metastatic breast cancer that overexpresses HER-2. Oncologist. 13:1114–1119. 2008. View Article : Google Scholar : PubMed/NCBI
21	Seidel J, Kunc K, Possinger K, Jehn C and Lüftner D: Effect of the tyrosine kinase inhibitor lapatinib on CUB-domain containing protein (CDCP1)-mediated breast cancer cell survival and migration. Biochem Biophys Res Commun. 414:226–232. 2011. View Article : Google Scholar : PubMed/NCBI
22	Zhu X, Wu L, Qiao H, Han T, Chen S, Liu X, et al: Autophagy stimulates apoptosis in HER2-overexpressing breast cancers treated by lapatinib. J Cell Biochem. Jun 21–2013.(Epub ahead of print). View Article : Google Scholar
23	Bian L, Wang T, Zhang S and Jiang Z: Trastuzumab plus capecitabine vs. lapatinib plus capecitabine in patients with trastuzumab resistance and taxane-pretreated metastatic breast cancer. Tumour Biol. 34:3153–3158. 2013. View Article : Google Scholar : PubMed/NCBI
24	Di Leo A, Gomez HL, Aziz Z, Zvirbule Z, Bines J, Arbushites MC, et al: Phase III, double-blind, randomized study comparing lapatinib plus paclitaxel with placebo plus paclitaxel as first-line treatment for metastatic breast cancer. J Clin Oncol. 26:5544–5552. 2008.PubMed/NCBI
25	Johnston S, Trudeau M, Kaufman B, Boussen H, Blackwell K, LoRusso P, et al: Phase II study of predictive biomarker profiles for response targeting human epidermal growth factor receptor 2 (HER-2) in advanced inflammatory breast cancer with lapatinib monotherapy. J Clin Oncol. 26:1066–1072. 2008. View Article : Google Scholar
26	Vandhana S, Coral K, Jayanthi U, Deepa PR and Krishnakumar S: Biochemical changes accompanying apoptotic cell death in retinoblastoma cancer cells treated with lipogenic enzyme inhibitors. Biochim Biophys Acta. 1831:1458–1466. 2013. View Article : Google Scholar : PubMed/NCBI
27	Liu ZL, Wang G, Peng AF, Luo QF, Zhou Y and Huang SH: Fatty acid synthase expression in osteosarcoma and its correlation with pulmonary metastasis. Oncol Lett. 4:878–882. 2012.PubMed/NCBI
28	Liu ZL, Mao JH, Peng AF, Yin QS, Zhou Y, Long XH and Huang SH: Inhibition of fatty acid synthase suppresses osteosarcoma cell invasion and migration via downregulation of the PI3K/Akt signaling pathway in vitro. Mol Med Rep. 7:608–612. 2013.PubMed/NCBI
29	Yoon S, Lee MY, Park SW, Moon JS, Koh YK, Ahn YH, et al: Up-regulation of acetyl-CoA carboxylase α and fatty acid synthase by human epidermal growth factor receptor 2 at the translational level in breast cancer cells. J Biol Chem. 282:26122–26131. 2007.
30	Li N, Bu X, Wu P, Wu P and Huang P: The ‘HER2-PI3K/Akt-FASN Axis’ regulated malignant phenotype of colorectal cancer cells. Lipids. 47:403–411. 2012.
31	Puig T, Turrado C, Benhamú B, Aguilar H, Relat J, Ortega-Gutiérrez S, et al: Novel inhibitors of fatty acid synthase with anticancer activity. Clin Cancer Res. 15:7608–7615. 2009. View Article : Google Scholar : PubMed/NCBI
32	Vazquez-Martin A, Colomer R, Brunet J, Lupu R and Menendez JA: Overexpression of fatty acid synthase gene activates HER1/HER2 tyrosine kinase receptors in human breast epithelial cells. Cell Prolif. 41:59–85. 2008. View Article : Google Scholar : PubMed/NCBI
33	Wood ER, Truesdale AT, McDonald OB, Yuan D, Hassell A, Dickerson SH, et al: A unique structure for epidermal growth factor receptor bound to GW572016 (Lapatinib): relationships among protein conformation, inhibitor off-rate, and receptor activity in tumor cells. Cancer Res. 64:6652–6659. 2004. View Article : Google Scholar
34	Xia W, Mullin RJ, Keith BR, Liu LH, Ma H, Rusnak DW, et al: Anti-tumor activity of GW572016: a dual tyrosine kinase inhibitor blocks EGF activation of EGFR/erbB2 and downstream Erk1/2 and AKT pathways. Oncogene. 21:6255–6263. 2002. View Article : Google Scholar : PubMed/NCBI