EGF upregulates RFPL3 and hTERT via the MEK signaling pathway in non‑small cell lung cancer cells

Lin,Chuan; Qin,Yu; Zhang,Hua; Gao,Ming‑Yang; Wang,Yan‑Fu

doi:10.3892/or.2018.6417

EGF upregulates RFPL3 and hTERT via the MEK signaling pathway in non‑small cell lung cancer cells

Authors:
- Chuan Lin
- Yu Qin
- Hua Zhang
- Ming‑Yang Gao
- Yan‑Fu Wang
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Affiliations: Department of Respiratory Medicine, The First Affiliated Hospital of Dalian Medical University, Dalian, Liaoning 116000, P.R. China, Department of Geriatrics, The First Affiliated Hospital of Dalian Medical University, Dalian, Liaoning 116000, P.R. China, Department of Geriatrics, The First Affiliated Hospital of Dalian Medical University, Dalian, Liaoning 116000, P.R. China, Department of Dermatology, The First Affiliated Hospital of Dalian Medical University, Dalian, Liaoning 116000, P.R. China
Published online on: May 8, 2018 https://doi.org/10.3892/or.2018.6417
Pages: 29-38
Copyright: © Lin et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Activation of the epidermal growth factor receptor (EGFR) during tumor development can trigger the MEK signaling pathway. In the present study, we investigated the MEK signaling pathway in non‑small cell lung cancer (NSCLC) cells with respect to the effect of epidermal growth factor (EGF) on expression of Ret finger protein like 3 (RFPL3) and human telomerase reverse transcriptase (hTERT), and the effect of RFPL3 overexpression on other MEK signaling proteins. In vitro, A549 and H1299 cells were treated with different concentrations of EGF for 24 h or 48 h. Expression of RFPL3 and hTERT at the mRNA and protein levels was determined by real‑time quantitative PCR (RT‑qPCR) and western blot analysis; cell viability was detected by MTT assay, and apoptosis was assayed via flow cytometry. We also pretreated A549 and H1299 cells with EGFR tyrosine kinase inhibitors, AG1478 and erlotinib, and MEK‑specific inhibitor (PD98059) in the presence of EGF. We used western blot analysis to assess the expression levels of RFPL3, hTERT and related MEK‑pathway proteins in A549 and H1299 cells transfected with RFPL3‑overexpression plasmids. EGF significantly upregulated RFPL3 and hTERT protein levels in the NSCLC cells. RFPL3 and hTERT proteins upregulation by EGF were attenuated by pretreatment with AG1478 and erlotinib. EGF promoted proliferation and inhibited apoptosis; PD98059 decreased RFPL3 and hTERT protein expression; and RFPL3 overexpression increased the expression of hTERT and related MEK‑pathway proteins. EGF upregulated RFPL3 and hTERT protein expression in NSCLC cells via the MEK pathway, promoted cell proliferation and inhibited apoptosis. RFPL3 overexpression increased expression of hTERT and related MEK signaling proteins (Ras, Raf, ERK and p‑ERK), which implies that RFPL3 is a potential therapeutic target for NSCLC.

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1	World Health Organization, . International Agency for Research on Cancer. Press Release No. 224. Lyon/London: 3–February. 2014
2	Minna JD and Schiller JH: Lung Cancer: Harrison's Principles of Internal Medicine (17th edition). 551–562. 2008.
3	Goldstraw P, Crowley J, Chansky K, Giroux DJ, Groome PA, Rami-Porta R, Postmus PE, Rusch V and Sobin L; and International Association for the Study of Lung Cancer International Staging Committee; Participating Institutions, : The IASLC lung cancer staging project: Proposals for the revision of the TNM stage groupings in the forthcoming (seventh) edition of the TNM classification of malignant tumours. J Thorac Oncol. 2:706–714. 2007. View Article : Google Scholar : PubMed/NCBI
4	DeSantis CE, Lin CC, Mariotto AB, Siegel RL, Stein KD, Kramer JL, Alteri R, Robbins AS and Jemal A: Cancer treatment and survivorship statistics, 2014. CA Cancer J Clin. 64:252–271. 2014. View Article : Google Scholar : PubMed/NCBI
5	Hirata Y, Moore GW, Bertagna C and Orth DN: Plasma concentrations of immunoreactive human epidermal growth factor (urogastrone) in man. J Clin Endocrinol Metab. 50:440–444. 1980. View Article : Google Scholar : PubMed/NCBI
6	Xiqing Li, Zunlan Zhao, Mingyue Liu, et al: Epidermal growth factor affects the growth and proliferation of non-small cell lung cancer A549 and H23 cells. Chinese Journal of applied diagnosis and treatment. 29:562–564. 2015.(In Chinese).
7	Bethune G, Bethune D, Ridgway N and Xu Z: Epidermal growth factor receptor (EGFR) in lung cancer: An overview and update. J Thorac Dis. 2:48–51. 2010.PubMed/NCBI
8	Lee HJ, Xu X, Choe G, Chung DH, Seo JW, Lee JH, Lee CT, Jheon S, Sung SW and Chung JH: Protein overexpression and gene amplification of epidermal growth factor receptor in nonsmall cell lung carcinomas: Comparison of four commercially available antibodies by immunohistochemistry and fluorescence in situ hybridization study. Lung Cancer. 68:375–382. 2010. View Article : Google Scholar : PubMed/NCBI
9	Hsu CP, Lee LW, Tang SC, Hsin IL, Lin YW and Ko JL: Epidermal growth factor activates telomerase activity by direct binding of Ets-2 to hTERT promoter in lung cancer cells. Tumour Biol. 36:5389–5398. 2015. View Article : Google Scholar : PubMed/NCBI
10	Chen W, Lu J, Qin Y, Wang J, Tian Y, Shi D, Wang S, Xiao Y, Dai M, Liu L, et al: Ret finger protein-like 3 promotes tumor cell growth by activating telomerase reverse transcriptase expression in human lung cancer cells. Oncotarget. 5:11909–11923. 2014.PubMed/NCBI
11	Mitsudomi T and Yatabe Y: Mutations of the epidermal growth factor receptor gene and related genes as determinants of epidermal growth factor receptor tyrosine kinase inhibitors sensitivity in lung cancer. Cancer Sci. 98:1817–1824. 2007. View Article : Google Scholar : PubMed/NCBI
12	Vivanco I and Sawyers CL: The phosphatidylinositol 3-Kinase AKT pathway in human cancer. Nat Rev Cancer. 2:489–501. 2002. View Article : Google Scholar : PubMed/NCBI
13	Murray SA, Yang S, Demicco E, Ying H, Sherr DH, Hafer LJ, Rogers AE, Sonenshein GE and Xiao ZX: Increased expression of MDM2, cyclin D1, and p27^Kip1 in carcinogen-induced rat mammary tumors. J Cell Biochem. 95:875–884. 2005. View Article : Google Scholar : PubMed/NCBI
14	Anderson D, Koch CA, Grey L, Ellis C, Moran MF and Pawson T: Binding of SH2 domains of phospholipase C gamma 1, GAP, and Src to activated growth factor receptors. Science. 250:979–982. 1990. View Article : Google Scholar : PubMed/NCBI
15	Yuqin Hao and Huixia Zheng: The mechanism research of MAPK/ERK signaling pathways in cancer treatment. China J Leprosy and Skin Diseases. 7:490–493. 2012.(In Chinese).
16	Roberts PJ and Stinchcombe TE: KRAS mutation: Should we test for it, and does it matter? J Clin Oncol. 31:1112–1121. 2013. View Article : Google Scholar : PubMed/NCBI
17	Loboda A, Nebozhyn M, Klinghoffer R, Frazier J, Chastain M, Arthur W, Roberts B, Zhang T, Chenard M, Haines B, et al: A gene expression signature of RAS pathway dependence predicts response to PI3K and RAS pathway inhibitors and expands the population of RAS pathway activated tumors. BMC Med Genomics. 3:262010. View Article : Google Scholar : PubMed/NCBI
18	Matallanas D and Crespo P: New druggable targets in the Ras pathway? Curr Opin Mol Ther. 12:674–683. 2010.PubMed/NCBI
19	Bonnefont J, Nikolaev SI, Perrier AL, Guo S, Cartier L, Sorce S, Laforge T, Aubry L, Khaitovich P, Peschanski M, et al: Evolutionary forces shape the human RFPL1,2,3 genes toward a role in neocortex development. Am J Hum Genet. 83:208–218. 2008. View Article : Google Scholar : PubMed/NCBI
20	Tan BH and Suzuki Y, Takahashi H, Ying PH, Takahashi C, Han Q, Chin WX, Chao SH, Sawasaki T, Yamamoto N and Suzuki Y: Identification of RFPL3 protein as a novel E3 ubiquitin ligase modulating the integration activity of human immunodeficiency virus, type 1 preintegration complex using a microtiter plate-based assay. J Biol Chem. 289:26368–26382. 2014. View Article : Google Scholar : PubMed/NCBI