lncRNA NEAT1 regulates gastric carcinoma cell proliferation, invasion and apoptosis via the miR‑500a‑3p/XBP‑1 axis

Zhou,Yun; Sha,Zhenghong; Yang,Yong; Wu,Shuimei; Chen,Hong

doi:10.3892/mmr.2021.12142

lncRNA NEAT1 regulates gastric carcinoma cell proliferation, invasion and apoptosis via the miR‑500a‑3p/XBP‑1 axis

Authors:
- Yun Zhou
- Zhenghong Sha
- Yong Yang
- Shuimei Wu
- Hong Chen
View Affiliations

Affiliations: Department of Gastroenterology, Zhongda Hospital Southeast University, Medical College of Southeast University, Nanjing, Jiangsu 210000, P.R. China, Department of General Surgery, Wuhu No. 1 People's Hospital, Wuhu, Anhui 241000, P.R. China, Department of Gastroenterology, Wuhu No. 1 People's Hospital, Wuhu, Anhui 241000, P.R. China
Published online on: May 9, 2021 https://doi.org/10.3892/mmr.2021.12142
Article Number: 503
Copyright: © Zhou et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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

Gastric cancer is a serious malignant tumor. Despite progression in gastric cancer research in recent years, the specific molecular mechanism underlying the pathogenesis of the disease is not completely understood. Long non‑coding RNA (lncRNA) nuclear paraspeckle assembly transcript 1 (NEAT1) affects the proliferation and metastasis of multiple types of tumor cells in colorectal cancer and breast cancer but its specific role in gastric cancer requires further investigation. The aim of the present study was to analyze the role of NEAT1 in gastric cancer. The expression of endoplasmic reticulum stress marker proteins and apoptosis‑related proteins in gastric cancer tissue and cell lines was analyzed using western blotting. The targeting relationship of NEAT1 and miR‑500a‑3p was analyzed using dual‑luciferase reporter assay. Cell proliferation was analyzed using CCK8 assay and colony formation assay while cell invasion was detected using Transwell assay. Cell apoptosis was analyzed using TUNEL staining and LC3 expression through immunofluorescent staining (IF). The results showed that lncRNA NEAT1‑overexpression gastric cancer cells were established to determine its effects on cell proliferation, invasion, apoptosis, autophagy and endoplasmic reticulum stress. Subsequently, microRNA (miR)‑500a was overexpressed in lncRNA NEAT1‑overexpression cells. Compared with the vector group, lncRNA NEAT1 overexpression significantly inhibited gastric cancer cell proliferation and invasion, but significantly promoted cell apoptosis. Furthermore, the results indicated that lncRNA NEAT1 targeted and downregulated the expression of miR‑500a‑3p, and miR‑500a‑3p targeted X‑box binding protein‑1 (XBP‑1) mRNA. lncRNA NEAT1 overexpression‑mediated inhibition of gastric cancer cell proliferation and invasion was significantly reversed by miR‑500a‑3p overexpression. Furthermore, compared with the vector group, the expression levels of endoplasmic reticulum stress‑related proteins (XBP‑1S/XBP‑1U ratio and 78‑kDa glucose‑regulated protein) and apoptosis‑related proteins (Bax and cleaved‑caspase‑3) were significantly upregulated by lncRNA NEAT1 overexpression; however, miR‑500a‑3p overexpression reversed lncRNA NEAT1 overexpression‑mediated effects on protein expression. The present study demonstrated that lncRNA NEAT1 inhibited gastric cancer cell proliferation and invasion, and promoted apoptosis by regulating the miR‑500a‑3p/XBP‑1 axis.

View Figures

View References

1	Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J and Jemal A: Global cancer statistics, 2012. CA Cancer J Clin. 65:87–108. 2015. View Article : Google Scholar : PubMed/NCBI
2	Chen W, Zheng R, Baade PD, Zhang S, Zeng H, Bray F, Jemal A, Yu XQ and He J: Cancer statistics in China, 2015. CA Cancer J Clin. 66:115–132. 2016. View Article : Google Scholar : PubMed/NCBI
3	Li R, Liu B and Gao J: The application of nanoparticles in diagnosis and theranostics of gastric cancer. Cancer Lett. 386:123–130. 2017. View Article : Google Scholar : PubMed/NCBI
4	Rona KA, Schwameis K, Zehetner J, Samakar K, Green K, Samaan J, Sandhu K, Bildzukewicz N, Katkhouda N and Lipham JC: Gastric cancer in the young: An advanced disease with poor prognostic features. J Surg Oncol. 115:371–375. 2017. View Article : Google Scholar : PubMed/NCBI
5	Saito Y, Li L, Coyaud E, Luna A, Sander C, Raught B, Asara JM, Brown M and Muthuswamy SK: LLGL2 rescues nutrient stress by promoting leucine uptake in ER⁺ breast cancer. Nature. 569:275–279. 2019. View Article : Google Scholar : PubMed/NCBI
6	Travis WD, Brambilla E and Riely GJ: New pathologic classification of lung cancer: Relevance for clinical practice and clinical trials. J Clin Oncol. 31:992–1001. 2013. View Article : Google Scholar : PubMed/NCBI
7	Mohamed E, Cao Y and Rodriguez PC: Endoplasmic reticulum stress regulates tumor growth and anti-tumor immunity: A promising opportunity for cancer immunotherapy. Cancer Immunol Immunother. 66:1069–1078. 2017. View Article : Google Scholar : PubMed/NCBI
8	Hu H, Tian M, Ding C and Yu S: The C/EBP homologous protein (CHOP) transcription factor functions in endoplasmic reticulum stress-induced apoptosis and microbial infection. Front Immunol. 9:30832019. View Article : Google Scholar : PubMed/NCBI
9	Urra H, Dufey E, Avril T, Chevet E and Hetz C: Endoplasmic reticulum stress and the hallmarks of cancer. Trends Cancer. 2:252–262. 2016. View Article : Google Scholar : PubMed/NCBI
10	Duplan E, Giaime E, Viotti J, Sévalle J, Corti O, Brice A, Ariga H, Qi L, Checler F and Alves da Costa C: ER-stress-associated functional link between Parkin and DJ-1 via a transcriptional cascade involving the tumor suppressor p53 and the spliced X-box binding protein XBP-1. J Cell Sci. 126:2124–2133. 2013. View Article : Google Scholar : PubMed/NCBI
11	Hsu SK, Chiu CC, Dahms HU, Chou CK, Cheng CM, Chang WT, Cheng KC, Wang HD and Lin IL: Unfolded protein response (UPR) in survival, dormancy, immunosuppression, metastasis, and treatments of cancer cells. Int J Mol Sci. 20:25182019. View Article : Google Scholar : PubMed/NCBI
12	Ding LH, Ye QN, Lu QJ, Zhu JH, Yan JH, Wang ZH and Huang CF: Expression of XBP-1 in breast cancer cell lines and its role in ERalpha signaling. Yi Chuan Xue Bao. 31:380–384. 2004.(In Chinese). PubMed/NCBI
13	Zhang L, Ding Y, Yuan Z, Liu J, Sun J, Lei F, Wu S, Li S and Zhang D: MicroRNA-500 sustains nuclear factor-κB activation and induces gastric cancer cell proliferation and resistance to apoptosis. Oncotarget. 6:2483–2495. 2015. View Article : Google Scholar : PubMed/NCBI
14	Li JH, Liu S, Zhou H, Qu LH and Yang JH: starBase v2.0: Decoding miRNA-ceRNA, miRNA-ncRNA and protein-RNA interaction networks from large-scale CLIP-Seq data. Nucleic Acids Res. 42((Database Issue)): D92–D97. 2014. View Article : Google Scholar : PubMed/NCBI
15	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
16	Sepúlveda D, Barrera MJ, Castro I, Aguilera S, Carvajal P, Lagos C, González S, Albornoz N, Bahamondes V, Quest AF, et al: Impaired IRE1α/XBP-1 pathway associated to DNA methylation might contribute to salivary gland dysfunction in Sjögren's syndrome patients. Rheumatology (Oxford). 57:1021–1032. 2018. View Article : Google Scholar
17	Chern YJ, Wong JCT, Cheng GSW, Yu A, Yin Y, Schaeffer DF, Kennecke HF, Morin G and Tai IT: The interaction between SPARC and GRP78 interferes with ER stress signaling and potentiates apoptosis via PERK/eIF2α and IRE1α/XBP-1 in colorectal cancer. Cell death Dis. 10:5042019. View Article : Google Scholar : PubMed/NCBI
18	Siegel R, Naishadham D and Jemal A: Cancer statistics, 2012. CA Cancer J Clin. 62:10–29. 2012. View Article : Google Scholar : PubMed/NCBI
19	Bertuccio P, Chatenoud L, Levi F, Praud D, Ferlay J, Negri E, Malvezzi M and La Vecchia C: Recent patterns in gastric cancer: A global overview. Int J Cancer. 125:666–673. 2009. View Article : Google Scholar : PubMed/NCBI
20	Wang M, Gu H, Qian H, Zhu W, Zhao C, Zhang X, Tao Y, Zhang L and Xu W: miR-17-5p/20a are important markers for gastric cancer and murine double minute 2 participates in their functional regulation. Eur J Cancer. 49:2010–2021. 2013. View Article : Google Scholar : PubMed/NCBI
21	Yu HG, Ai YW, Yu LL, Zhou XD, Liu J, Li JH, Xu XM, Liu S, Chen J, Liu F, et al: Phosphoinositide 3-kinase/Akt pathway plays an important role in chemoresistance of gastric cancer cells against etoposide and doxorubicin induced cell death. Int J Cancer. 122:433–443. 2008. View Article : Google Scholar : PubMed/NCBI
22	Kopp F and Mendell JT: Functional classification and experimental dissection of long noncoding RNAs. Cell. 172:393–407. 2018. View Article : Google Scholar : PubMed/NCBI
23	Zhang M, Weng W, Zhang Q, Wu Y, Ni S, Tan C, Xu M, Sun H, Liu C, Wei P and Du X: The lncRNA NEAT1 activates Wnt/β-catenin signaling and promotes colorectal cancer progression via interacting with DDX5. J Hematol Oncol. 11:1132018. View Article : Google Scholar : PubMed/NCBI
24	Zhao Y and Wang Y and Wang Y: Up-regulated miR-500a enhances hepatocarcinoma metastasis by repressing PTEN expression. Biosci Rep. 37:BSR201708372017. View Article : Google Scholar : PubMed/NCBI
25	Guo Y, Chen L, Sun C and Yu C: MicroRNA-500a promotes migration and invasion in hepatocellular carcinoma by activating the Wnt/β-catenin signaling pathway. Biomed Pharmacother. 91:13–20. 2017. View Article : Google Scholar : PubMed/NCBI
26	Cubillos-Ruiz JR, Bettigole SE and Glimcher LH: Tumorigenic and immunosuppressive effects of endoplasmic reticulum stress in cancer. Cell. 168:692–706. 2017. View Article : Google Scholar : PubMed/NCBI
27	Oakes SA and Papa FR: The role of endoplasmic reticulum stress in human pathology. Annu Rev Pathol. 10:173–194. 2015. View Article : Google Scholar : PubMed/NCBI
28	Zhu XM, Yao FH, Yao YM, Dong N, Yu Y and Sheng ZY: Endoplasmic reticulum stress and its regulator XBP-1 contributes to dendritic cell maturation and activation induced by high mobility group box-1 protein. Int J Biochem Cell Biol. 44:1097–1105. 2012. View Article : Google Scholar : PubMed/NCBI
29	Barez SR, Atar AM and Aghaei M: Mechanism of inositol-requiring enzyme 1-alpha inhibition in endoplasmic reticulum stress and apoptosis in ovarian cancer cells. J Cell Commun Signal. 14:403–415. 2020. View Article : Google Scholar : PubMed/NCBI
30	Mowers EE, Sharifi MN and Macleod KF: Functions of autophagy in the tumor microenvironment and cancer metastasis. FEBS J. 285:1751–1766. 2018. View Article : Google Scholar : PubMed/NCBI
31	Lopez J and Tait SW: Mitochondrial apoptosis: Killing cancer using the enemy within. Br J Cancer. 112:957–962. 2015. View Article : Google Scholar : PubMed/NCBI
32	Bao L, Zhang M, Han S, Zhan Y, Guo W, Teng F, Liu F, Guo M, Zhang L, Ding G and Wang Q: MicroRNA-500a promotes the progression of hepatocellular carcinoma by post-transcriptionally targeting BID. Cell Physiol Biochem. 47:2046–2055. 2018. View Article : Google Scholar : PubMed/NCBI
33	Liu Z, Su D, Qi X and Ma J: MiR-500a-5p promotes glioblastoma cell proliferation, migration and invasion by targeting chromodomain helicase DNA binding protein 5. Mol Med Rep. 18:2689–2696. 2018.PubMed/NCBI
34	Larsen AC: Conjunctival malignant melanoma in Denmark: Epidemiology, treatment and prognosis with special emphasis on tumorigenesis and genetic profile. Acta Ophthalmol. 94:1–27. 2016. View Article : Google Scholar : PubMed/NCBI
35	Liao XH, Xie Z and Guan CN: MiRNA-500a-3p inhibits cell proliferation and invasion by targeting lymphocyte antigen 6 complex locus K (LY6K) in human non-small cell lung cancer. Neoplasma. 65:673–682. 2018. View Article : Google Scholar : PubMed/NCBI
36	Davies MP, Barraclough DL, Stewart C, Joyce KA, Eccles RM, Barraclough R, Rudland PS and Sibson DR: Expression and splicing of the unfolded protein response gene XBP-1 are significantly associated with clinical outcome of endocrine-treated breast cancer. Int J Cancer. 123:85–88. 2008. View Article : Google Scholar : PubMed/NCBI
37	Hsu HT, Hsing MT, Yeh CM, Chen CJ, Yang JS and Yeh KT: Decreased cytoplasmic X-box binding protein-1 expression is associated with poor prognosis and overall survival in patients with oral squamous cell carcinoma. Clin Chim Acta. 479:66–71. 2018. View Article : Google Scholar : PubMed/NCBI