shRNA‑mediated silencing of TARBP2 inhibits NCI‑H1299 non‑small cell lung cancer cell invasion and migration via the JNK/STAT3/AKT pathway

Shi,Yue; Zuo,Duo; Wang,Xia; Han,Meng; Wu,Yan

doi:10.3892/mmr.2016.5723

shRNA‑mediated silencing of TARBP2 inhibits NCI‑H1299 non‑small cell lung cancer cell invasion and migration via the JNK/STAT3/AKT pathway

Authors:
- Yue Shi
- Duo Zuo
- Xia Wang
- Meng Han
- Yan Wu
View Affiliations

Affiliations: Department of Clinical Laboratory, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Tianjin's Clinical Research Center for Cancer and Key Laboratory of Cancer Prevention and Therapy, Tianjin 300060, P.R. China, Department of Gastrointestinal Oncology, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Tianjin's Clinical Research Center for Cancer and Key Laboratory of Cancer Prevention and Therapy, Tianjin 300060, P.R. China, Department of Clinical Laboratory, The First Center Hospital, Tianjin 300192, P.R. China, Department of Clinical Laboratory, TEAD Community Health Service Center, Tianjin 300457, P.R. China
Published online on: September 6, 2016 https://doi.org/10.3892/mmr.2016.5723
Pages: 3725-3730

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

Metastasis is a major cause of lung cancer-associated mortality. The current study aimed to investigate the effects and mechanisms of TAR (human immunodeficiency virus 1) RNA binding protein 2 (TARBP2) in the invasion and migration of non‑small cell lung cancer in vitro. The highly metastatic cell clone H1299/M02 was obtained by TARBP2 overexpression. Expression of TARBP2 in H1299/M02 was also downregulated to different levels via small hairpin RNAs (shRNAs). Subsequent to TARBP2 silencing, the proliferation of H1299/M02 cells was predominantly unaffected, while invasion and migration were significantly inhibited. A positive correlation was observed between invasion and migration and the level of TARBP2 silencing in vitro. Western blotting and reverse transcription‑quantitative polymerase chain reaction indicated that the protein expression levels of amyloid β (A4) precursor protein (APP) and zinc finger protein 395 (ZNF395) were upregulated, while expression levels of pro‑metastatic proteins including interleukin (IL)‑1β, IL‑8, cyclooxygenase (COX)‑2, matrix metalloproteinase 2 (MMP2) and MMP9 were downregulated. Phosphorylation of c‑Jun N‑terminal kinase (JNK), signal transducer and activator of transcription 3 (STAT3) and protein kinase B (AKT) were also inhibited. Overexpression of TARBP2 was suggested to be involved in the metastasis of H1299/M02 cells. Silencing of TARBP2 was able to upregulate levels of APP and ZNF395, in addition to inhibiting metastasis‑promoting cytokines, the JNK/STAT3/AKT pathway and COX‑2 to attenuate the invasion and migration of cancer cells.

View Figures

View References

1	Dela Cruz CS, Tanoue LT and Matthay RA: Lung cancer: epidemiology, etiology and prevention. Clin Chest Med. 32:605–644. 2011. View Article : Google Scholar : PubMed/NCBI
2	Spano JP, Chouahnia K and Morère JF: Cyclooxygenase 2 inhibitors and lung carcinoma. Bull Cancer. 91(Suppl 2): S109–S112. 2004.In French.
3	D'Addario G, Früh M, Reck M, Baumann P, Klepetko W and Felip E; ESMO Guidelines Working Group: Metastatic non-small-cell lung cancer: ESMO clinical practice guidelines for diagnosis, treatment and follow-up. Ann Oncol. 21(Suppl 5): v116–v119. 2010. View Article : Google Scholar : PubMed/NCBI
4	Gupta GP and Massagué J: Cancer metastasis: Building a framework. Cell. 127:679–695. 2006. View Article : Google Scholar : PubMed/NCBI
5	Gatignol A, Buckler-White A, Berkhout B and Jeang KT: Characterization of a human TAR RNA-binding protein that activates the HIV-1 LTR. Science. 251:1597–1600. 1991. View Article : Google Scholar : PubMed/NCBI
6	Gatignol A, Buckler C and Jeang KT: Relatedness of an RNA-binding motif in human immunodeficiency virus type 1 TAR RNA-binding protein TRBP to human P1/dsI kinase and Drosophila staufen. Mol Cell Biol. 13:2193–2202. 1993. View Article : Google Scholar : PubMed/NCBI
7	Chendrimada TP, Gregory RI, Kumaraswamy E, Norman J, Cooch N, Nishikura K and Shiekhattar R: TRBP recruits the Dicer complex to Ago2 for microRNA processing and gene silencing. Nature. 436:740–744. 2005. View Article : Google Scholar : PubMed/NCBI
8	Goodarzi H, Zhang S, Buss CG, Fish L, Tavazoie S and Tavazoie SF: Metastasis-suppressor transcript destabilization through TARBP2 binding of mRNA hairpins. Nature. 513:256–260. 2014. View Article : Google Scholar : PubMed/NCBI
9	Zhu Y, Zhang X, Qi L, Cai Y, Yang P, Xuan G and Jiang Y: HULC long noncoding RNA silencing suppresses angiogenesis by regulating ESM-1 via the PI3K/Akt/mTOR signaling pathway in human gliomas. Oncotarget. 12:14429–14440. 2016.
10	Kuhn PH, Wang H, Dislich B, Colombo A, Zeitschel U, Ellwart JW, Kremmer E, Rossner S and Lichtenthaler SF: ADAM10 is the physiologically relevant, constitutive alpha-secretase of the amyloid precursor protein in primary neurons. EMBO J. 29:3020–3032. 2010. View Article : Google Scholar : PubMed/NCBI
11	Pastuszak-Lewandoska D, Domańska D, Czarnecka KH, Kordiak J, Migdalska-Sęk M, Nawrot E, Kiszałkiewicz J, Antczak A, Górski P and Brzeziańska E: Expression of STAT5, COX-2 and PIAS3 in correlation with NSCLC histhopathological features. PLoS One. 9:e1042652014. View Article : Google Scholar : PubMed/NCBI
12	Fan X, Liu Y, Jiang J, Ma Z, Wu H, Liu T, Liu M, Li X and Tang H: miR-20a promotes proliferation and invasion by targeting APP in human ovarian cancer cells. Acta Biochim Biophys Sin (Shanghai). 42:318–324. 2010. View Article : Google Scholar
13	Gough M, Blanthorn-Hazell S, Delury C and Parkin E: The E1 copper binding domain of full-length amyloid precursor protein mitigates copper-induced growth inhibition in brain metastatic prostate cancer DU145 cells. Biochem Biophys Res Commun. 453:741–747. 2014. View Article : Google Scholar : PubMed/NCBI
14	Miyazaki T, Ikeda K, Horie-Inoue K and Inoue S: Amyloid precursor protein regulates migration and metalloproteinase gene expression in prostate cancer cells. Biochem Biophys Res Commun. 452:828–833. 2014. View Article : Google Scholar : PubMed/NCBI
15	Tanaka K, Shouguchi-Miyata J, Miyamoto N and Ikeda JE: Novel nuclear shuttle proteins, HDBP1 and HDBP2, bind to neuronal cell-specific cis-regulatory element in the promoter for the human Huntington's disease gene. J Biol Chem. 279:7275–7286. 2004. View Article : Google Scholar
16	Pang F, Zha R, Zhao Y, Wang Q, Chen D, Zhang Z, Chen T, Yao M, Gu J and He X: MiR-525-3p enhances the migration and invasion of liver cancer cells by downregulating ZNF395. PLoS One. 9:e908672014. View Article : Google Scholar : PubMed/NCBI
17	Yano S, Nokihara H, Yamamoto A, Goto H, Ogawa H, Kanematsu T, Miki T, Uehara H, Saijo Y, Nukiwa T and Sone S: Multifunctional interleukin-1beta promotes metastasis of human lung cancer cells in SCID mice via enhanced expression of adhesion-, invasion- and angiogenesis-related molecules. Cancer Sci. 94:244–252. 2003. View Article : Google Scholar : PubMed/NCBI
18	Cheng D, Kong H and Li Y: Prognostic values of VEGF and IL-8 in malignant pleural effusion in patients with lung cancer. Biomarkers. 18:386–390. 2013. View Article : Google Scholar : PubMed/NCBI
19	Cheng CY, Hsieh HL, Sun CC, Lin CC, Luo SF and Yang CM: IL-1 beta induces urokinase-plasminogen activator expression and cell migration through PKC alpha, JNK1/2 and NF-kappaB in A549 cells. J Cell Physiol. 219:183–193. 2009. View Article : Google Scholar
20	Carmi Y, Rinott G, Dotan S, Elkabets M, Rider P, Voronov E and Apte RN: Microenvironment-derived IL-1 and IL-17 interact in the control of lung metastasis. J Immunol. 186:3462–3471. 2011. View Article : Google Scholar : PubMed/NCBI
21	Ryan BM, Pine SR, Chaturvedi AK, Caporaso N and Harris CC: A combined prognostic serum interleukin-8 and interleukin-6 classifier for stage 1 lung cancer in the prostate, lung, colorectal, and ovarian cancer screening trial. J Thorac Oncol. 10:1494–1503. 2014. View Article : Google Scholar
22	Iizasa T, Fujisawa T, Suzuki M, Motohashi S, Yasufuku K, Yasukawa T, Baba M and Shiba M: Elevated levels of circulating plasma matrix metalloproteinase 9 in non-small cell lung cancer patients. Clin Cancer Res. 5:149–153. 1999.PubMed/NCBI
23	Kessenbrock K, Plaks V and Werb Z: Matrix metalloproteinases: Regulators of the tumor microenvironment. Cell. 141:52–67. 2010. View Article : Google Scholar : PubMed/NCBI
24	Shiraga M1, Yano S, Yamamoto A, Ogawa H, Goto H, Miki T, Miki K, Zhang H and Sone S: Organ heterogeneity of host-derived matrix metalloproteinase expression and its involvement in multiple-organ metastasis by lung cancer cell lines. Cancer Res. 62:5967–5973. 2002.PubMed/NCBI
25	Jacobs EJ, Hsing AW, Bain EB, Stevens VL, Wang Y, Chen J, Chanock SJ, Zheng SL, Xu J and Thun MJ: Polymorphisms in angiogenesis-related genes and prostate cancer. Cancer Epidemiol Biomarkers Prev. 17:972–977. 2008. View Article : Google Scholar : PubMed/NCBI
26	Jiao Y, Sun KK, Zhao L, Xu JY, Wang LL and Fan SJ: Suppression of human lung cancer cell proliferation and metastasis in vitro by the transducer of ErbB-2.1 (TOB1). Acta Pharmacol Sin. 33:250–260. 2012. View Article : Google Scholar