Increased MIR31HG lncRNA expression increases gefitinib resistance in non-small cell lung cancer cell lines through the EGFR/PI3K/AKT signaling pathway

Wang,Bing; Jiang,Hong; Wang,Limin; Chen,Xueqin; Wu,Kan; Zhang,Shirong; Ma,Shenglin; Xia,Bing

doi:10.3892/ol.2017.5878

Increased MIR31HG lncRNA expression increases gefitinib resistance in non-small cell lung cancer cell lines through the EGFR/PI3K/AKT signaling pathway

Authors:
- Bing Wang
- Hong Jiang
- Limin Wang
- Xueqin Chen
- Kan Wu
- Shirong Zhang
- Shenglin Ma
- Bing Xia
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Affiliations: Department of Radiation Oncology, Hangzhou First People's Hospital, Hangzhou Cancer Hospital, Hangzhou, Zhejiang 310002, P.R. China, Department of Thoracic Surgery, Hangzhou First People's Hospital, Hangzhou, Zhejiang 310002, P.R. China, Department of Respiration, Hangzhou First People's Hospital, Hangzhou, Zhejiang 310002, P.R. China, Department of Medical Oncology, Hangzhou First People's Hospital, Hangzhou, Zhejiang 310002, P.R. China
Published online on: March 20, 2017 https://doi.org/10.3892/ol.2017.5878
Pages: 3494-3500
Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The aim of the present study was to gain insight into the molecular mechanism of gefitinib resistance in non-small cell lung cancer (NSCLC), and demonstrate whether long noncoding RNA (lncRNA) expression signatures differ between gefitinib‑sensitive PC9 and gefitinib-resistant PC9 (PC9‑R) cell lines. PC9 and PC9‑R cells were treated with gefitinib and, after 48 h, proliferation and apoptosis were analyzed using a Cell Counting Kit‑8 (CCK‑8) assay and flow cytometry. Microarray expression profiling of lncRNAs was undertaken in both PC9 and PC9‑R cells, and the expression profiles were verified by reverse transcription quantitative‑polymerase chain reaction. The EGFR/PI3K/AKT signaling pathway and mitochondrial apoptosis protein expression levels were assessed by western blot analysis. The PC9 cell line treated with gefitinib had a more significant effect on cell viability and apoptosis than the PC9‑R cell line (P<0.05). Expression of various lncRNAs differed significantly between the two cell lines, and MIR31HG expression in particular was significantly higher in PC9‑R cells. As expected, MIR31HG lncRNA knockdown sensitized PC9‑R cells to gefitinib, and further experiments revealed that turning off the EGFR/PI3K/AKT signaling pathway activated expression of p53 in PC9‑R cells transfected with si‑MIR31HG. Furthermore, PC9‑R cells transfected with si‑MIR31HG induced cell apoptosis through the mitochondrial apoptosis pathway, and arrested the cell cycle in the G0/G1 phase. The results of the current study suggest that MIR31HG lncRNA levels in PC9‑R cells are higher than in PC9 cells. Furthermore, overexpression of MIR31HG lncRNAs may contribute to gefitinib resistance in PC9‑R cells through the EGFR/PI3K/AKT pathway, which impacts on cell proliferation, apoptosis and the cell cycle. MIR31HG lncRNA may therefore be a novel candidate biomarker for future therapeutic strategies involving EGFR-TKIs.

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1	Siegel R, Ma J, Zou Z and Jemal A: Cancer statistics, 2014. CA Cancer J Clin. 64:9–29. 2014. View Article : Google Scholar : PubMed/NCBI
2	Cruz CS Dela, Tanoue LT and Matthay RA: Lung cancer: Epidemiology, etiology, and prevention. Clin Chest Med. 32:605–644. 2011. View Article : Google Scholar : PubMed/NCBI
3	Hotta K, Kiura K, Ueoka H, Tabata M, Fujiwara K, Kozuki T, Okada T, Hisamoto A and Tanimoto M: Effect of gefitinib (‘Iressa’, ZD1839) on brain metastases in patients with advanced non-small-cell lung cancer. Lung Cancer. 46:255–261. 2004. View Article : Google Scholar : PubMed/NCBI
4	Burotto M, Manasanch EE, Wilkerson J and Fojo T: Gefitinib and erlotinib in metastatic non-small cell lung cancer: A meta-analysis of toxicity and efficacy of randomized clinical trials. Oncologist. 20:400–410. 2015. View Article : Google Scholar : PubMed/NCBI
5	Engelman JA and Settleman J: Acquired resistance to tyrosine kinase inhibitors during cancer therapy. Curr Opin Genet Dev. 18:73–79. 2008. View Article : Google Scholar : PubMed/NCBI
6	Prabhakar CN: Epidermal growth factor receptor in non-small cell lung cancer. Transl Lung Cancer Res. 4:110–118. 2015.PubMed/NCBI
7	Chen J, Wang W, Wang H, Liu X and Guo X: Combination treatment of ligustrazine piperazine derivate DLJ14 and adriamycin inhibits progression of resistant breast cancer through inhibition of the EGFR/PI3K/Akt survival pathway and induction of apoptosis. Drug Discov Ther. 8:33–41. 2014. View Article : Google Scholar : PubMed/NCBI
8	Ørom UA, Derrien T, Beringer M, Gumireddy K, Gardini A, Bussotti G, Lai F, Zytnicki M, Notredame C, Huang Q, et al: Long noncoding RNAs with enhancer-like function in human cells. Cell. 143:46–58. 2010. View Article : Google Scholar : PubMed/NCBI
9	Cui Z, Ren S, Lu J, Wang F, Xu W, Sun Y, Wei M, Chen J, Gao X, Xu C, et al: The prostate cancer-up-regulated long noncoding RNA PlncRNA-1 modulates apoptosis and proliferation through reciprocal regulation of androgen receptor. Urol Oncol. 31:1117–1123. 2013. View Article : Google Scholar : PubMed/NCBI
10	Yang F, Zhang L, Huo XS, Yuan JH, Xu D, Yuan SX, Zhu N, Zhou WP, Yang GS, Wang YZ, et al: Long noncoding RNA high expression in hepatocellular carcinoma facilitates tumor growth through enhancer of zeste homolog 2 in humans. Hepatology. 54:1679–1689. 2011. View Article : Google Scholar : PubMed/NCBI
11	Ji P, Diederichs S, Wang W, Böing S, Metzger R, Schneider PM, Tidow N, Brandt B, Buerger H, Bulk E, et al: MALAT-1, a novel noncoding RNA and thymosin beta4 predict metastasis and survival in early-stage non-small cell lung cancer. Oncogene. 22:8031–8041. 2003. View Article : Google Scholar : PubMed/NCBI
12	Liu XH, Liu ZL, Sun M, Liu J, Wang ZX and De W: The long non-coding RNA HOTAIR indicates a poor prognosis and promotes metastasis in non-small cell lung cancer. BMC Cancer. 13:4642013. View Article : Google Scholar : PubMed/NCBI
13	Sun M, Liu XH, Wang KM, Nie FQ, Kong R, Yang JS, Xia R, Xu TP, Jin FY, Liu ZJ, et al: Down regulation of BRAF activated non-coding RNA is associated with poor prognosis for non-small cell lung cancer and promotes metastasis by affecting epithelialmesenchymal transition. Mol Cancer. 13:682014. View Article : Google Scholar : PubMed/NCBI
14	Han L, Kong R, Yin DD, Zhang EB, Xu TP, De W and Shu YQ: Low expression of long noncoding RNA GAS6-AS1 predicts a poor prognosis in patients with NSCLC. Med Oncol. 30:6942013. View Article : Google Scholar : PubMed/NCBI
15	Zhang L, Zhou XF, Pan GF and Zhao JP: Enhanced expression of long non-coding RNA ZXF1 promoted the invasion and metastasis in lung adenocarcinoma. Biomed Pharmacother. 68:401–407. 2014. View Article : Google Scholar : PubMed/NCBI
16	Ricciuti B, Mecca C, Crinò L, Baglivo S, Cenci M and Metro G: Non-coding RNAs in lung cancer. Oncoscience. 1:674–705. 2014. View Article : Google Scholar : PubMed/NCBI
17	Thomas D and Livak KJ: Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc. 3:1101–1108. 2008. View Article : Google Scholar : PubMed/NCBI
18	Rahman AF, Korashy HM and Kassem MG: Gefitinib. Profiles Drug Subst Excip Relat Methodol. 39:239–264. 2014. View Article : Google Scholar : PubMed/NCBI
19	Pilkington G, Boland A, Brown T, Oyee J, Bagust A and Dickson R: A systematic review of the clinical effectiveness of first-line chemotherapy for adult patients with locally advanced or metastatic non-small cell lung cancer. Thorax. 70:359–367. 2015. View Article : Google Scholar : PubMed/NCBI
20	Yuan Y, Li XF, Chen JQ, Dong CX, Weng SS and Huang JJ: Critical appraisal of the role of gefitinib in the management of locally advanced or metastatic non-small cell lung cancer. Onco Targets Ther. 7:841–852. 2014. View Article : Google Scholar : PubMed/NCBI
21	Engelman JA and Jänne PA: Mechanisms of acquired resistance to epidermal growth factor receptor tyrosine kinase inhibitors in non-small cell lung cancer. Clin Cancer Res. 14:2895–2899. 2008. View Article : Google Scholar : PubMed/NCBI
22	Engelman JA, Zejnullahu K, Mitsudomi T, Song Y, Hyland C, Park JO, Lindeman N, Gale CM, Zhao X, Christensen J, et al: MET amplification leads to gefitinib resistance in lung cancer by activating ERBB3 signaling. Science. 316:1039–1043. 2007. View Article : Google Scholar : PubMed/NCBI
23	Zhang J, Zhu N and Chen X: A novel long noncoding RNA LINC01133 is upregulated in lung squamous cell cancer and predicts survival. Tumour Biol. 36:7465–7471. 2015. View Article : Google Scholar : PubMed/NCBI
24	Cai H, Chen J, He B, Li Q, Li Y and Gao Y: A FOXM1 related long non-coding RNA contributes to gastric cancer cell migration. Mol Cell Biochem. 406:31–41. 2015. View Article : Google Scholar : PubMed/NCBI
25	Gao Y, Chen G, Zeng Y, Zeng J, Lin M, Liu X and Liu J: Invasion and metastasis-related long noncoding RNA expression profiles in hepatocellular carcinoma. Tumour Biol. 36:7409–7422. 2015. View Article : Google Scholar : PubMed/NCBI
26	Ren S, Wang F, Shen J, Sun Y, Xu W, Lu J, Wei M, Xu C, Wu C, Zhang Z, et al: Long non-coding RNA metastasis associated in lung adenocarcinoma transcript1 derived miniRNA as a novel plasma-based biomarker for diagnosing prostate cancer. Eur J Cancer. 49:2949–2959. 2013. View Article : Google Scholar : PubMed/NCBI
27	Qiu M, Xu Y, Yang X, Wang J, Hu J, Xu L and Yin R: CCAT2 is a lung adenocarcinoma specific long non-coding RNA and promotes invasion of non-small cell lung cancer. Tumour Biol. 35:5375–5380. 2014. View Article : Google Scholar : PubMed/NCBI
28	Gutschner T, Hämmerle M, Eissmann M, Hsu J, Kim Y, Hung G, Revenko A, Arun G, Stentrup M, Gross M, et al: The noncoding RNA MALAT1 is a critical regulator of the metastasis phenotype of lung cancer cells. Cancer Res. 73:1180–1189. 2013. View Article : Google Scholar : PubMed/NCBI
29	Cheng N, Li X, Zhao C, Ren S, Chen X, Cai W, Zhao M, Zhang Y, Li J, Wang Q and Zhou C: Microarray expression profile of long non-coding RNAs in EGFR-TKIs resistance of human non-small cell lung cancer. Oncol Rep. 33:833–839. 2015.PubMed/NCBI
30	Nguyen KS, Kobayashi S and Costa DB: Acquired resistance to epidermal growth factor receptor tyrosine kinase inhibitorsin non-small-cell lung cancers dependent on the epidermal growth factor receptor pathway. Clin Lung Cancer. 10:281–289. 2009. View Article : Google Scholar : PubMed/NCBI
31	Li H, Schmid-Bindert G, Wang D, Zhao Y, Yang X, Su B and Zhou C: Blocking the PI3K/AKT and MEK/ERK signaling pathways can overcome gefitinib-resistance in non-small cell lung cancer cell lines. Adv Med Sci. 56:275–284. 2011. View Article : Google Scholar : PubMed/NCBI
32	Kang XH, Xu ZY, Gong YB, Wang LF, Wang ZQ, Xu L, Cao F and Liao MJ: Bufalin reverses HGF-induced resistance to EGFR-TKIs in EGFR mutant lung cancer cells via blockage of Met/PI3k/Akt pathway and induction of apoptosis. Evid Based Complement Alternat Med. 2013:2438592013. View Article : Google Scholar : PubMed/NCBI
33	Sordella R, Bell DW, Haber DA and Settleman J: Gefitinib-sensitizing EGFR mutations in lung cancer activate anti-apoptotic pathways. Science. 305:1163–1167. 2004. View Article : Google Scholar : PubMed/NCBI
34	Qiao M, Sheng S and Pardee AB: Metastasis and AKT activation. Cell Cycle. 7:2991–2996. 2008. View Article : Google Scholar : PubMed/NCBI
35	Slack A, Chen Z, Tonelli R, Pule M, Hunt L, Pession A and Shohet JM: The p53 regulatory gene MDM2 is a direct transcriptional target of MYCN in neuroblastoma. Proc Natl Acad Sci USA. 18:731–736. 2005. View Article : Google Scholar
36	Harris MH and Thompson CB: The role of the Bcl-2 family in the regulation of outer mitochondrial membrane permeability. Cell Death Differ. 7:1182–1191. 2000. View Article : Google Scholar : PubMed/NCBI
37	Elkholi R, Renault TT, Serasinghe MN and Chipuk JE: Putting the pieces together: How is the mitochondrial pathway of apoptosis regulated in cancer and chemotherapy? Cancer Metab. 2:162014. View Article : Google Scholar : PubMed/NCBI
38	Muthu M, Cheriyan VT and Rishi AK: CARP-1/CCAR1: A biphasic regulator of cancer cell growth and apoptosis. Oncotarget. 6:6499–6510. 2015. View Article : Google Scholar : PubMed/NCBI