Effect of TW37 on the growth of H1975 EGFR‑TKI‑resistant lung cancer cells and its underlying mechanisms

Liu,Jixian; Yao,Xinhuang; Wu,Da; Ye,Yiwang; Wu,Qiang; Liu,Suyue; Wu,Hao

doi:10.3892/mmr.2017.8181

Effect of TW37 on the growth of H1975 EGFR‑TKI‑resistant lung cancer cells and its underlying mechanisms

Authors:
- Jixian Liu
- Xinhuang Yao
- Da Wu
- Yiwang Ye
- Qiang Wu
- Suyue Liu
- Hao Wu
View Affiliations

Affiliations: Department of Thoracic Surgery, Peking University Shenzhen Hospital, Shenzhen, Guangdong 518036, P.R. China, Central Laboratory of Peking University Shenzhen Hospital, Shenzhen, Guangdong 518036, P.R. China, Department of Thoracic Surgery, Anhui Medical University Shenzhen Clinical College, Shenzhen, Guangdong 518036, P.R. China
Published online on: November 29, 2017 https://doi.org/10.3892/mmr.2017.8181
Pages: 2509-2514

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

Previous studies have suggested that the B‑cell lymphoma 2 (Bcl‑2) inhibitor, TW37, may induce apoptosis of the non‑small cell lung cancer cell line, H1975/epidermal growth factor receptor‑tyrosine kinase inhibitor (EGFR‑TKI), which exhibits secondary resistance to EGFR‑TKI. However, the effects of TW37 on H1975/EGFR‑TKI cells remain unclear. The aim of the present study was to investigate the effects of TW37 on the growth of H1975/EGFR‑TKI cells and explore the underlying mechanisms. An in vitro study was performed, whereby H1975/EGFR‑TKI cells were treated with serially increasing concentrations of TW37. MTT, flow cytometry, migration and transwell invasion assays were preformed to investigate the proliferation, apoptosis, migration and invasion of H1975/EGFR‑TKI cells, respectively. In addition, reverse transcription‑polymerase chain reaction and western blot analyses were performed to detect the mRNA and protein expression levels of apoptosis‑associated factors, respectively. An enzyme‑linked immunosorbent assay was performed to detect phosphatidylinositol [3,4,5] tris‑phosphate (PIP3) expression. The results suggested that the mRNA and protein expression levels of Bcl‑2 were significantly decreased in TW37‑treated cells when compared with the untreated control group. Following treatment with TW37, the proliferation, migration and invasion ability of H1975/EGFR‑TKI cells decreased in a dose‑dependent manner, while the percentage of apoptotic cells increased. In addition, the results demonstrated that TW37 reduced the expression of PIP3 and the phosphorylation of AKT serine/threonine kinase 1 (AKT) in H1975/EGFR‑TKI cells in a dose‑dependent manner. In conclusion, TW37 inhibited H1975/EGFR‑TKI cell growth and induced cell apoptosis potentially via suppression of AKT signaling pathway activation.

View Figures

View References

1	Révész D, Engelhardt EG, Tamminga JJ, Schramel FMNH, Onwuteaka-Philipsen BD, van de Garde EMW, Steyerberg EW, Jansma EP, De Vet HCW and Coupé VMH: Decision support systems for incurable non-small cell lung cancer: A systematic review. BMC Med Inform Decis Mak. 17:17–144. 2017. View Article : Google Scholar : PubMed/NCBI
2	Yang L, Parkin DM, Li LD, Chen YD and Bray F: Estimation and projection of the national profile of cancer mortality in China: 1991–2010. Br J Cancer. 90:2157–2166. 2004. View Article : Google Scholar : PubMed/NCBI
3	Swami U, Shah U and Goel S: Eribulin in non-small cell lung cancer: Challenges and potential strategies. Expert Opin Investig Drugs. 26:495–508. 2017. View Article : Google Scholar : PubMed/NCBI
4	Berhoune M, Banu E, Scotte F, Prognon P, Oudard S and Bonan B: Therapeutic strategy for treatment of metastatic non-small cell lung cancer. Ann Pharmacother. 42:1640–1652. 2008. View Article : Google Scholar : PubMed/NCBI
5	Zhang L, Li J, Hu J, Li D, Wang X, Zhang R, Zhang H, Shi M and Chen H: Cigarette smoke extract induces EGFR-TKI resistance via promoting EGFR signaling pathway and ROS generation in NSCLC cell lines. Lung Cancer. 109:109–116. 2017. View Article : Google Scholar : PubMed/NCBI
6	Inoue A, Kobayashi K, Maemondo M, Sugawara S, Oizumi S, Isobe H, Gemma A, Harada M, Yoshizawa H, Kinoshita I, et al: Updated overall survival results from a randomized phase III trial comparing gefitinib with carboplatin-paclitaxel for chemo-naïve non-small cell lung cancer with sensitive EGFR gene mutations (NEJ002). Ann Oncol. 24:54–59. 2013. View Article : Google Scholar : PubMed/NCBI
7	Mitsudomi T, Morita S, Yatabe Y, Negoro S, Okamoto I, Tsurutani J, Seto T, Satouchi M, Tada H, Hirashima T, et al: Gefitinib versus cisplatin plus docetaxel in patients with non-small-cell lung cancer harbouring mutations of the epidermal growth factor receptor (WJTOG3405): An open label, randomised phase 3 trial. Lancet Oncol. 11:121–128. 2010. View Article : Google Scholar : PubMed/NCBI
8	Rosell R, Carcereny E, Gervais R, Vergnenegre A, Massuti B, Felip E, Palmero R, Garcia-Gomez R, Pallares C, Sanchez JM, et al: Erlotinib versus standard chemotherapy as first-line treatment for European patients with advanced EGFR mutation-positive non-small-cell lung cancer (EURTAC): A multicentre, open-label, randomised phase 3 trial. Lancet Oncol. 13:239–246. 2012. View Article : Google Scholar : PubMed/NCBI
9	Sequist LV, Yang JC, Yamamoto N, O'Byrne K, Hirsh V, Mok T, Geater SL, Orlov S, Tsai CM, Boyer M, et al: Phase III study of afatinib or cisplatin plus pemetrexed in patients with metastatic lung adenocarcinoma with EGFR mutations. J Clin Oncol. 31:3327–3334. 2013. View Article : Google Scholar : PubMed/NCBI
10	Pao W and Chmielecki J: Rational, biologically based treatment of EGFR-mutant non-small-cell lung cancer. Nat Rev Cancer. 10:760–774. 2010. View Article : Google Scholar : PubMed/NCBI
11	Su JC, Lin KL, Chien CM, Tseng CH, Chen YL, Chang LS and Lin SR: Naphtho[1,2-b]furan-4,5-dione inactivates EGFR and PI3K/Akt signaling pathways in human lung adenocarcinoma A549 cells. Life Sci. 86:207–213. 2010. View Article : Google Scholar : PubMed/NCBI
12	Wang Z, Azmi AS, Ahmad A, Banerjee S, Wang S, Sarkar FH and Mohammad RM: TW-37, a small-molecule inhibitor of Bcl-2, inhibits cell growth and induces apoptosis in pancreatic cancer: Involvement of Notch-1 signaling pathway. Cancer Res. 69:2757–2765. 2009. View Article : Google Scholar : PubMed/NCBI
13	Ashimori N, Zeitlin BD, Zhang Z, Warner K, Turkienicz IM, Spalding AC, Teknos TN, Wang S and Nör JE: TW-37, a small-molecule inhibitor of Bcl-2, mediates S-phase cell cycle arrest and suppresses head and neck tumor angiogenesis. Mol Cancer Ther. 8:893–903. 2009. View Article : Google Scholar : PubMed/NCBI
14	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
15	Oltersdorf T, Elmore SW, Shoemaker AR, Armstrong RC, Augeri DJ, Belli BA, Bruncko M, Deckwerth TL, Dinges J, Hajduk PJ, et al: An inhibitor of Bcl-2 family proteins induces regression of solid tumours. Nature. 435:677–681. 2005. View Article : Google Scholar : PubMed/NCBI
16	Lee HW, Choi YW, Han JH, Kim JH, Jung JH, Jeong SH, Kang SY, Choi JH, Oh YT, Park KJ, et al: Expression of excision repair cross-complementation group 1 protein predicts poor outcome in advanced non-small cell lung cancer patients treated with platinum-based doublet chemotherapy. Lung Cancer. 65:377–382. 2012. View Article : Google Scholar
17	Qiao GB, Zeng WS, Peng LJ, et al: Overexpression of bcl-2 and sensitivity to Platinum agents in non small cell lung cancer. J Clin Oncol. 24 Suppl:410S–411S. 2013.(In Chinese).
18	Ma J, Zhu W and Zhou Q: Expression and Significance of bag-1, bcl-2 in Non-small cell lung cancer and the correlation with Multi-drug resistance. Zhongguo Fei Ai Za Zhi. 12:1089–1094. 2009.(In Chinese). PubMed/NCBI
19	Koty PP, Zhang H, Franklin WA, Yousem SA, Landreneau R and Levitt ML: In vivo expression of p53 and Bcl-2 and their role in programmed cell death in premalignant and malignant lung lesions. Lung Cancer. 35:155–163. 2012. View Article : Google Scholar
20	Su JC, Lin KL, Chien CM, Tseng CH, Chen YL, Chang LS and Lin SR: Naphtho[1,2-b]furan-4,5-dione inactivates EGFR and PI3K/Akt signaling pathways in human lung adenocarcinoma A549 cells. Life Sci. 86:207–213. 2010. View Article : Google Scholar : PubMed/NCBI
21	Song JH, Kandasamy K, Zemskova M, Lin YW and Kraft AS: The BH3 mimetic ABT-737 induces cancer cell senescence. Cancer Res. 71:506–515. 2011. View Article : Google Scholar : PubMed/NCBI
22	Zeitlin BD, Joo E, Dong Z, Warner K, Wang G, Nikolovska-Coleska Z, Wang S and Nör JE: Antiangiogenic effect of TW37, a small-molecule inhibitor of Bcl-2. Cancer Res. 66:8698–8706. 2006. View Article : Google Scholar : PubMed/NCBI
23	Zeitlin BD, Spalding AC, Campos MS, Ashimori N, Dong Z, Wang S, Lawrence TS and Nör JE: Metronomic small molecule inhibitor of Bcl-2 (TW-37) is antiangiogenic and potentiates the anti-tumor effect of ionizing radiation. Int J Radiat Oncol Biol Phys. 78:879–887. 2010. View Article : Google Scholar : PubMed/NCBI
24	Wang H, Zhang Z, Wei X and Dai R: Small-molecule inhibitor of Bcl-2 (TW-37) suppresses growth and enhances cisplatin-induced apoptosis in ovarian cancer cells. J Ovarian Res. 8:32015. View Article : Google Scholar : PubMed/NCBI
25	Matsuzawa A, Nishitoh H, Tobiume K, Takeda K and Ichijo H: Physiological roles of ASK1-mediated signal transduction in oxidative stress- and endoplasmic reticulum stress-induced apoptosis: Advanced findings from ASK1 knockout mice. Antioxid Redox Signal. 4:415–425. 2002. View Article : Google Scholar : PubMed/NCBI
26	Vogler M, Walter HS and Dyer MJS: Targeting anti-apoptotic BCL2 family proteins in haematological malignancies-from pathogenesis to treatment. Br J Haematol. 178:364–379. 2017. View Article : Google Scholar : PubMed/NCBI
27	Wang Z, Song W, Aboukammad M, Mohammad M, Wang G, Banerjee S, Kong D, Wang S, Sarkar FH and Mohammad RM: TW-37, a small-molecule inhibitor of Bcl-2, inhibits cell growth and invasion in pancreatic cancer. Int J Cancer. 123:958–966. 2008. View Article : Google Scholar : PubMed/NCBI
28	Zheng Q, Wang B, Gao J, Xin N, Wang W, Song X, Shao Y and Zhao C: CD155 knockdown promotes apoptosis via AKT/Bcl-2/Bax in colon cancer cells. J Cell Mol Med. 16–Aug;2017.(Epub ahead of print). View Article : Google Scholar
29	Liang Z, Wang X, Xu X, Xie B, Ji A, Meng S, Li S, Zhu Y, Wu J, Hu Z, et al: MicroRNA-608 inhibits proliferation of bladder cancer via AKT/FOXO3a signaling pathway. Mol Cancer. 16:962017. View Article : Google Scholar : PubMed/NCBI