The expression, significance and function of cancer susceptibility candidate 9 in lung squamous cell carcinoma: A bioinformatics and in vitro investigation

Gao,Li; Guo,Yi‑Nan; Zeng,Jiang‑Hui; Ma,Fu‑Chao; Luo,Jie; Zhu,Hua‑Wei; Xia,Shuang; Wei,Kang‑Lai; Chen,Gang

doi:10.3892/ijo.2019.4758

The expression, significance and function of cancer susceptibility candidate 9 in lung squamous cell carcinoma: A bioinformatics and in vitro investigation

Authors:
- Li Gao
- Yi‑Nan Guo
- Jiang‑Hui Zeng
- Fu‑Chao Ma
- Jie Luo
- Hua‑Wei Zhu
- Shuang Xia
- Kang‑Lai Wei
- Gang Chen
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Affiliations: Department of Pathology, First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi 530021, P.R. China, Department of Medical Oncology, First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi 530021, P.R. China, Department of Medical Oncology, Second Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi 530021, P.R. China, Department of Human Anatomy, Guangxi Medical University, Nanning, Guangxi 530021, P.R. China
Published online on: March 19, 2019 https://doi.org/10.3892/ijo.2019.4758
Pages: 1651-1664
Copyright: © Gao et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The cancer susceptibility candidate 9 (CASC9) gene has been reported to exert an oncogenic effect in several types of cancer. However, its role in lung squamous cell carcinoma (LUSC) is unknown. Therefore, the present study examined the expression of CASC9 in LUSC and non‑cancer tissues by reverse transcription‑quantitative polymerase chain reaction assays and by data mining of high‑throughput public databases, including The Cancer Genome Atlas, the Gene Expression Omnibus, ArrayExpress and the Cancer Cell Line Encyclopedia. In vitro experiments were conducted to investigate the effects of CASC9 on the viability and the proliferation of LUSC cells. Furthermore, consulting the alteration status of CASC9 in LUSC from cBioPortal, functional enrichment analysis of co‑expressed genes, prediction of potential transcription factors, and inspection of adjacent protein‑coding genes were conducted to explore the potential molecular mechanism of CASC9 in LUSC. The results revealed that CASC9 was overexpressed in LUSC tissue, and significantly associated with the malignant progression of LUSC. In vitro experiments demonstrated that CASC9 knockdown by RNA interference attenuated the viability and proliferation of LUSC cells. Multiple copies of CASC9 gene were detected in 4 of 179 available sequenced LUSC cases. A functional enrichment analysis of 200 co‑expressed genes indicated that these genes were significantly associated with terms, including ‘cell‑cell junction organization’, ‘desmosome organization’, ‘epidermis development’, ‘Hippo signaling pathway’, ‘pathogenic Escherichia coli infection’ and ‘PID HIF1 TF pathway’. Three genes, Fos‑related antigen 2 (FOSL2), SWI/SNF complex subunit SMARCC2, and transcription factor COE1 (EBF1), were predicted by lncRNAMap to be associated with CASC9. Among these, the expression of FOSL2 and EBF1 was positively and negatively correlated with the expression of CASC9, respectively. Two adjacent protein‑coding genes, cysteine‑rich secretory protein LCCL domain‑containing 1 and hepatocyte nuclear factor 4‑γ, were also positively correlated with CASC9 expression. In conclusion, the present data suggest that CASC9 serves as an oncogene in LUSC and may be a promising target for alternative therapeutic options for patients with this condition.

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1	Perez-Moreno P, Brambilla E, Thomas R and Soria JC: Squamous cell carcinoma of the lung: molecular subtypes and therapeutic opportunities. Clin Cancer Res. 18:2443–2451. 2012. View Article : Google Scholar : PubMed/NCBI
2	Guo NL, Dowlati A, Raese RA, Dong C, Chen G, Beer DG, Shaffer J, Singh S, Bokhary U, Liu L, et al: A predictive 7-gene assay and prognostic protein biomarkers for non-small cell lung cancer. EBioMedicine. 32:102–110. 2018. View Article : Google Scholar : PubMed/NCBI
3	Siegel RL, Miller KD and Jemal A: Cancer statistics, 2018. CA Cancer J Clin. 68:7–30. 2018. View Article : Google Scholar : PubMed/NCBI
4	Yan X, Shen H, Jiang H, Hu D, Wang J and Wu X: YXQ-EQ induces apoptosis and inhibits signaling pathways important for metastasis in non-small cell lung carcinoma cells. Cell Physiol Biochem. 49:911–919. 2018. View Article : Google Scholar : PubMed/NCBI
5	Wu C, Li X, Zhang D, Xu B, Hu W, Zheng X, Zhu D, Zhou Q, Jiang J and Wu C: IL-1beta-mediated up-regulation of WT1D via miR-144-3p and their synergistic effect with NF-kappaB/COX-2/HIF-1alpha pathway on cell proliferation in LUAD. Cell Physiol Biochem. 48:2493–2502. 2018. View Article : Google Scholar
6	Lee B, Lee T, Lee SH, Choi YL and Han J: Clinicopathologic characteristics of EGFR, KRAS, and ALK alterations in 6,595 lung cancers. Oncotarget. 7:23874–23884. 2016.PubMed/NCBI
7	Hou Z, Xu C, Xie H, Xu H, Zhan P, Yu L and Fang X: Long noncoding RNAs expression patterns associated with chemo response to cisplatin based chemotherapy in lung squamous cell carcinoma patients. PLoS One. 9:e1081332014. View Article : Google Scholar : PubMed/NCBI
8	Zhao J, Liu Y, Zhang W, Zhou Z, Wu J, Cui P, Zhang Y and Huang G: Long non-coding RNA Linc00152 is involved in cell cycle arrest, apoptosis, epithelial to mesenchymal transition, cell migration and invasion in gastric cancer. Cell Cycle. 14:3112–3123. 2015. View Article : Google Scholar : PubMed/NCBI
9	Fan CN, Ma L and Liu N: Systematic analysis of lncRNA-miRNA-mRNA competing endogenous RNA network identifies four-lncRNA signature as a prognostic biomarker for breast cancer. J Transl Med. 16:2642018. View Article : Google Scholar : PubMed/NCBI
10	Huang YK and Yu JC: Circulating microRNAs and long non-coding RNAs in gastric cancer diagnosis: An update and review. World J Gastroenterol. 21:9863–9886. 2015. View Article : Google Scholar : PubMed/NCBI
11	Wang X, Sehgal L, Jain N, Khashab T, Mathur R and Samaniego F: LncRNA MALAT1 promotes development of mantle cell lymphoma by associating with EZH2. J Transl Med. 14:3462016. View Article : Google Scholar : PubMed/NCBI
12	Dong L, Ding H, Li Y, Xue D and Liu Y: LncRNA TINCR is associated with clinical progression and serves as tumor suppressive role in prostate cancer. Cancer Manag Res. 10:2799–2807. 2018. View Article : Google Scholar : PubMed/NCBI
13	Liang X, Qi M, Wu R, Liu A, Chen D, Tang L, Chen J, Hu X, Li W, Zhan L, et al: Long non-coding RNA CUDR promotes malignant phenotypes in pancreatic ductal adenocarcinoma via activating AKT and ERK signaling pathways. Int J Oncol. 53:2671–2682. 2018.PubMed/NCBI
14	Cao W, Wu W, Shi F, Chen X, Wu L, Yang K, Tian F, Zhu M, Chen G, Wang W, et al: Integrated analysis of long noncoding RNA and coding RNA expression in esophageal squamous cell carcinoma. Int J Genomics. 2013:4805342013. View Article : Google Scholar : PubMed/NCBI
15	Su X, Li G and Liu W: The long noncoding RNA cancer susceptibility candidate 9 promotes nasopharyngeal carcinogenesis via stabilizing HIF1a. DNA Cell Biol. 36:394–400. 2017. View Article : Google Scholar : PubMed/NCBI
16	Shang C, Sun L, Zhang J, Zhao B, Chen X, Xu H and Huang B: Silence of cancer susceptibility candidate 9 inhibits gastric cancer and reverses chemoresistance. Oncotarget. 8:15393–15398. 2017. View Article : Google Scholar : PubMed/NCBI
17	Ma P, Zhang M, Nie F, Huang Z, He J, Li W and Han L: Transcriptome analysis of EGFR tyrosine kinase inhibitors resistance associated long noncoding RNA in non-small cell lung cancer. Biomed Pharmacother. 87:20–26. 2017. View Article : Google Scholar : PubMed/NCBI
18	Zhou J, Xiao H, Yang X, Tian H, Xu Z, Zhong Y, Ma L, Zhang W, Qiao G and Liang J: Long noncoding RNA CASC9.5 promotes the proliferation and metastasis of lung adenocarcinoma. Sci Rep. 8:372018. View Article : Google Scholar : PubMed/NCBI
19	Xiong DD, Li ZY, Liang L, He RQ, Ma FC, Luo DZ, Hu XH and Chen G: The lncRNA NEAT1 accelerates lung adenocarcinoma deterioration and binds to mir-193a-3pas a competitive endogenous RNA. Cell Physiol Biochem. 48:905–918. 2018. View Article : Google Scholar
20	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Δ Δ C(T)) Method. Methods. 25:402–408. 2001. View Article : Google Scholar
21	Tomczak K, Czerwinska P and Wiznerowicz M: The Cancer Genome Atlas (TCGA): An immeasurable source of knowledge. Contemp Oncol (Pozn). 19A:A68–A77. 2015.
22	Hutter C and Zenklusen JC: The Cancer Genome Atlas: Creating lasting value beyond its data. Cell. 173:283–285. 2018. View Article : Google Scholar : PubMed/NCBI
23	He RQ, Gao L, Ma J, Li ZY, Hu XH and Chen G: Oncogenic role of miR-183-5p in lung adenocarcinoma: A comprehensive study of qPCR, in vitro experiments and bioinformatic analysis. Oncol Rep. 40:83–100. 2018.PubMed/NCBI
24	Dang Y, Luo D, Rong M and Chen G: Underexpression of miR-34a in hepatocellular carcinoma and its contribution towards enhancement of proliferating inhibitory effects of agents targeting c-MET. PLoS One. 8:e610542013. View Article : Google Scholar : PubMed/NCBI
25	Rong M, Chen G and Dang Y: Increased miR-221 expression in hepatocellular carcinoma tissues and its role in enhancing cell growth and inhibiting apoptosis in vitro. BMC Cancer. 13:212013. View Article : Google Scholar : PubMed/NCBI
26	Huang S and He R: Synergistic effect of MiR-146a mimic and cetuximab on hepatocellular carcinoma cells. BioMed Research International. 2014:3841212014.PubMed/NCBI
27	Dang YW, Zeng J, He RQ, Rong MH, Luo DZ and Chen G: Effects of miR-152 on cell growth inhibition, motility suppression and apoptosis induction in hepatocellular carcinoma cells. Asian Pac J Cancer Prev. 15:4969–4976. 2014. View Article : Google Scholar : PubMed/NCBI
28	Gao J, Aksoy BA, Dogrusoz U, Dresdner G, Gross B, Sumer SO, Sun Y, Jacobsen A, Sinha R, Larsson E, et al: Integrative analysis of complex cancer genomics and clinical profiles using the cBio- Portal. Sci Signal. 6:pl 12013. View Article : Google Scholar
29	Ooi AT, Gower AC, Zhang KX, Vick JL, Hong L, Nagao B, Wallace WD, Elashoff DA, Walser TC, Dubinett SM, et al: Molecular profiling of premalignant lesions in lung squamous cell carcinomas identifies mechanisms involved in stepwise carcinogenesis. Cancer Prev Res (Phila). 7:487–495. 2014. View Article : Google Scholar
30	Arima C, Kajino T, Tamada Y, Imoto S, Shimada Y, Nakatochi M, Suzuki M, Isomura H, Yatabe Y, Yamaguchi T, et al: Lung adenocarcinoma subtypes definable by lung development-related miRNA expression profiles in association with clinicopathologic features. Carcinogenesis. 35:2224–2231. 2014. View Article : Google Scholar : PubMed/NCBI
31	Wu Y, Hu L, Liang Y, Li J, Wang K, Chen X, Meng H, Guan X, Yang K and Bai Y: Up-regulation of lncRNA CASC9 promotes esophageal squamous cell carcinoma growth by negatively regulating PDCD4 expression through EZH2. Mol Cancer. 16:1502017. View Article : Google Scholar : PubMed/NCBI
32	Lankat-Buttgereit B and Göke R: The tumour suppressor Pdcd4: Recent advances in the elucidation of function and regulation. Biol Cell. 101:309–317. 2009. View Article : Google Scholar : PubMed/NCBI
33	Allgayer H: Pdcd4, a colon cancer prognostic that is regulated by a microRNA. Crit Rev Oncol Hematol. 73:185–191. 2010. View Article : Google Scholar
34	Hwang SK, Minai-Tehrani A, Lim HT, Shin JY, An GH, Lee KH, Park KR, Kim YS, Beck GR Jr, Yang HS, et al: Decreased level of PDCD4 (programmed cell death 4) protein activated cell proliferation in the lung of A/J mouse. J Aerosol Med Pulm Drug Deliv. 23:285–293. 2010. View Article : Google Scholar : PubMed/NCBI
35	Kaneko S, Li G, Son J, Xu CF, Margueron R, Neubert TA and Reinberg D: Phosphorylation of the PRC2 component Ezh2 is cell cycle-regulated and up-regulates its binding to ncRNA. Genes Dev. 24:2615–2620. 2010. View Article : Google Scholar : PubMed/NCBI
36	Wang KC, Yang YW, Liu B, Sanyal A, Corces-Zimmerman R, Chen Y, Lajoie BR, Protacio A, Flynn RA, Gupta RA, et al: A long noncoding RNA maintains active chromatin to coordinate homeotic gene expression. Nature. 472:120–124. 2011. View Article : Google Scholar : PubMed/NCBI
37	Gao GD, Liu XY, Lin Y, Liu HF and Zhang GJ: LncRNA CASC9 promotes tumorigenesis by affecting EMT and predicts poor prognosis in esophageal squamous cell cancer. Eur Rev Med Pharmacol Sci. 22:422–429. 2018.PubMed/NCBI
38	Kostenko S, Dumitriu G and Moens U: Tumour promoting and suppressing roles of the atypical MAP kinase signalling pathway ERK3/4-MK5. J Mol Signal. 7:92012. View Article : Google Scholar : PubMed/NCBI
39	Hu Y, Yang C, Yang S, Cheng F, Rao J and Wang X: miR-665 promotes hepatocellular carcinoma cell migration, invasion, and proliferation by decreasing Hippo signaling through targeting PTPRB. Cell Death Dis. 9:9542018. View Article : Google Scholar : PubMed/NCBI
40	Wang J, Sun D, Wang Y, Ren F, Pang S, Wang D and Xu S: FOSL2 positively regulates TGF-ß1 signalling in non-small cell lung cancer. PLoS One. 9:e1121502014. View Article : Google Scholar
41	Remmelink M, Mijatovic T, Gustin A, Mathieu A, Rombaut K, Kiss R, Salmon I and Decaestecker C: Identification by means of cDNA microarray analyses of gene expression modifications in squamous non-small cell lung cancers as compared to normal bronchial epithelial tissue. Int J Oncol. 26:247–258. 2005.
42	Georgopoulos K: Ebf1 in DNA repair and leukemogenesis. Blood. 125:3969–3971. 2015. View Article : Google Scholar : PubMed/NCBI
43	Wang J, Zhang J, Xu L, Zheng Y, Ling D and Yang Z: Expression of HNF4G and its potential functions in lung cancer. Oncotarget. 9:18018–18028. 2017.
44	Jiang B, Li Y, Qu X, Zhu H, Tan Y, Fan Q, Jiang Y, Liao M and Wu X: Long noncoding RNA cancer susceptibility candidate 9 promotes doxorubicin-resistant breast cancer by binding to enhancer of zeste homolog 2. Int J Mol Med. 42:2801–2810. 2018.PubMed/NCBI
45	Klingenberg M, Groß M, Goyal A, Polycarpou-Schwarz M, Miersch T, Ernst AS, Leupold J, Patil N, Warnken U, Allgayer H, et al: The long noncoding RNA cancer susceptibility 9 and RNA binding protein heterogeneous nuclear ribonucleoprotein L form a complex and coregulate genes linked to AKT signaling. Hepatology. 68:1817–1832. 2018. View Article : Google Scholar : PubMed/NCBI
46	Liang Y, Chen X, Wu Y, Li J, Zhang S, Wang K, Guan X, Yang K and Bai Y: LncRNA CASC9 promotes esophageal squamous cell carcinoma metastasis through upregulating LAMC2 expression by interacting with the CREB-binding protein. Cell Death Differ. 25:1980–1995. 2018. View Article : Google Scholar : PubMed/NCBI