RNA-binding motif protein 5 inhibits the proliferation of cigarette smoke-transformed BEAS-2B cells through cell cycle arrest and apoptosis

Lv,Xue-Jiao; Du,Yan-Wei; Hao,Yu-Qiu; Su,Zhen-Zhong; Zhang,Lin; Zhao,Li-Jing; Zhang,Jie

doi:10.3892/or.2016.4551

RNA-binding motif protein 5 inhibits the proliferation of cigarette smoke-transformed BEAS-2B cells through cell cycle arrest and apoptosis

Authors:
- Xue-Jiao Lv
- Yan-Wei Du
- Yu-Qiu Hao
- Zhen-Zhong Su
- Lin Zhang
- Li-Jing Zhao
- Jie Zhang
View Affiliations

Affiliations: Department of Respiratory Medicine, The Second Affiliated Hospital of Jilin University, Changchun, Jilin 130041, P.R. China, Department of Pathology, College of Basic Medicine, Changchun University of Chinese Medicine, Changchun, Jilin 130117, P.R. China, Department of Pathophysiology, College of Basic Medicine, Jilin University, Changchun, Jilin 130021, P.R. China
Published online on: January 12, 2016 https://doi.org/10.3892/or.2016.4551
Pages: 2315-2327

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

Cigarette smoking has been shown to be the most significant risk factor for lung cancer. Recent studies have also indicated that RNA-binding motif protein 5 (RBM5) can modulate apoptosis and suppress tumor growth. The present study focused on the role of RBM5 in the regulation of cigarette smoke extract (CSE)-induced transformation of bronchial epithelial cells into the cancerous phenotype and its mechanism of action. Herein, we exposed normal BEAS-2B cells for 8 days to varying concentrations of CSE or dimethylsulfoxide (DMSO), followed by a recovery period of 2 weeks. Next, the RBM5 protein was overexpressed in these transformed BEAS-2B cells though lentiviral infection. Later, the morphological changes, cell proliferation, cell cycle, apoptosis, invasion and migration were assessed. In addition, we analyzed the role of RBM5 in xenograft growth. The expression of RBM5 along with the genes related to cell cycle regulation, apoptosis and invasion were also examined. Finally, our results revealed that BEAS-2B cells exposed to 100 µg/ml CSE acquired phenotypic changes and formed tumors in nude mice, indicative of their cancerous transformation and had reduced RBM5 expression. Subsequent overexpression of RBM5 in these cells significantly inhibited their proliferation, induced G1/S arrest, triggered apoptosis and inhibited their invasion and migration, including xenograft growth. Thus, we established an in vitro model of CSE-induced cancerous transformation and concluded that RBM5 overexpression inhibited the growth of these transformed cells through cell cycle arrest and induction of apoptosis. Therefore, our study suggests the importance of RBM5 in the pathogenesis of smoking-related cancer.

View Figures

View References

1	Chen M, Yang T, Meng X and Sun T: Azithromycin attenuates cigarette smoke extract-induced oxidative stress injury in human alveolar epithelial cells. Mol Med Rep. 11:3414–3422. 2015.
2	Lemjabbar-Alaoui H, Dasari V, Sidhu SS, Mengistab A, Finkbeiner W, Gallup M and Basbaum C: Wnt and Hedgehog are critical mediators of cigarette smoke-induced lung cancer. PLoS One. 1:e932006.
3	Sasco AJ, Secretan MB and Straif K: Tobacco smoking and cancer: A brief review of recent epidemiological evidence. Lung Cancer. 45(Suppl 2): S3–S9. 2004.
4	Oh JJ, Razfar A, Delgado I, Reed RA, Malkina A, Boctor B and Slamon DJ: 3p21.3 tumor suppressor gene H37/Luca15/RBM5 inhibits growth of human lung cancer cells through cell cycle arrest and apoptosis. Cancer Res. 66:3419–3427. 2006.
5	Mourtada-Maarabouni M, Sutherland LC, Meredith JM and Williams GT: Simultaneous acceleration of the cell cycle and suppression of apoptosis by splice variant delta-6 of the candidate tumour suppressor LUCA-15/RBM5. Genes Cells. 8:109–119. 2003.
6	Zhao L, Li R, Shao C, Li P, Liu J and Wang K: 3p21.3 tumor suppressor gene RBM5 inhibits growth of human prostate cancer PC-3 cells through apoptosis. World J Surg Oncol. 10:2472012.
7	Welling DB, Lasak JM, Akhmametyeva E, Ghaheri B and Chang LS: cDNA microarray analysis of vestibular schwannomas. Otol Neurotol. 23:736–748. 2002.
8	Oh JJ, West AR, Fishbein MC and Slamon DJ: A candidate tumor suppressor gene, H37, from the human lung cancer tumor suppressor locus 3p21.3. Cancer Res. 62:3207–3213. 2002.
9	Rintala-Maki ND, Goard CA, Langdon CE, Wall VE, Traulsen KE, Morin CD, Bonin M and Sutherland LC: Expression of RBM5-related factors in primary breast tissue. J Cell Biochem. 100:1440–1458. 2007.
10	Oh JJ, Grosshans DR, Wong SG and Slamon DJ: Identification of differentially expressed genes associated with HER-2/neu over-expression in human breast cancer cells. Nucleic Acids Res. 27:4008–4017. 1999.
11	Wang W, Cassidy J, O'Brien V, Ryan KM and Collie-Duguid E: Mechanistic and predictive profiling of 5-fluorouracil resistance in human cancer cells. Cancer Res. 64:8167–8176. 2004.
12	Sutherland LC, Wang K and Robinson AG: RBM5 as a putative tumor suppressor gene for lung cancer. J Thorac Oncol. 5:294–298. 2010.
13	Oh JJ, Taschereau EO, Koegel AK, Ginther CL, Rotow JK, Isfahani KZ and Slamon DJ: RBM5/H37 tumor suppressor, located at the lung cancer hot spot 3p21.3, alters expression of genes involved in metastasis. Lung Cancer. 70:253–262. 2010.
14	Akhtar MJ, Ahamed M, Khan MA, Alrokayan SA, Ahmad I and Kumar S: Cytotoxicity and apoptosis induction by nanoscale talc particles from two different geographical regions in human lung epithelial cells. Environ Toxicol. 29:394–406. 2014.
15	Niu Z, Jin W, Zhang L and Li X: Tumor suppressor RBM5 directly interacts with the DExD/H-box protein DHX15 and stimulates its helicase activity. FEBS Lett. 586:977–983. 2012.
16	Mourtada-Maarabouni M, Sutherland LC and Williams GT: Candidate tumour suppressor LUCA-15 can regulate multiple apoptotic pathways. Apoptosis. 7:421–432. 2002.
17	Mourtada-Maarabouni M, Keen J, Clark J, Cooper CS and Williams GT: Candidate tumor suppressor LUCA-15/RBM5/H37 modulates expression of apoptosis and cell cycle genes. Exp Cell Res. 312:1745–1752. 2006.
18	Bechara EG, Sebestyén E, Bernardis I, Eyras E and Valcárcel J: RBM5, 6, and 10 differentially regulate NUMB alternative splicing to control cancer cell proliferation. Mol Cell. 52:720–733. 2013.
19	Schamberger AC, Mise N, Jia J, Genoyer E, Yildirim AÖ, Meiners S and Eickelberg O: Cigarette smoke-induced disruption of bronchial epithelial tight junctions is prevented by transforming growth factor-β. Am J Respir Cell Mol Biol. 50:1040–1052. 2014.
20	Du H, Sun J, Chen Z, Nie J, Tong J and Li J: Cigarette smoke-induced failure of apoptosis resulting in enhanced neoplastic transformation in human bronchial epithelial cells. J Toxicol Environ Health A. 75:707–720. 2012.
21	Drabkin HA, West JD, Hotfilder M, Heng YM, Erickson P, Calvo R, Dalmau J, Gemmill RM and Sablitzky F: DEF-3 (g16/NY-LU-12), an RNA binding protein from the 3p21.3 homozygous deletion region in SCLC. Oncogene. 18:2589–2597. 1999.
22	Maarabouni MM and Williams GT: The antiapoptotic RBM5/LUCA-15/H37 gene and its role in apoptosis and human cancer: Research update. ScientificWorldJournal. 6:1705–1712. 2006.
23	Liang H, Zhang J, Shao C, Zhao L, Xu W, Sutherland LC and Wang K: Differential expression of RBM5, EGFR and KRAS mRNA and protein in non-small cell lung cancer tissues. J Exp Clin Cancer Res. 31:362012.
24	Kobayashi T, Ishida J, Musashi M, Ota S, Yoshida T, Shimizu Y, Chuma M, Kawakami H, Asaka M, Tanaka J, et al: p53 transactivation is involved in the antiproliferative activity of the putative tumor suppressor RBM5. Int J Cancer. 128:304–318. 2011.
25	Satyanarayana A and Kaldis P: Mammalian cell-cycle regulation: Several Cdks, numerous cyclins and diverse compensatory mechanisms. Oncogene. 28:2925–2939. 2009.
26	Hochegger H, Takeda S and Hunt T: Cyclin-dependent kinases and cell-cycle transitions: Does one fit all? Nat Rev Mol Cell Biol. 9:910–916. 2008.
27	Fung TK and Poon RY: A roller coaster ride with the mitotic cyclins. Semin Cell Dev Biol. 16:335–342. 2005.
28	Hedberg Y, Ljungberg B, Roos G and Landberg G: Retinoblastoma protein in human renal cell carcinoma in relation to alterations in G1/S regulatory proteins. Int J Cancer. 109:189–193. 2004.
29	Vermeulen K, Van Bockstaele DR and Berneman ZN: The cell cycle: A review of regulation, deregulation and therapeutic targets in cancer. Cell Prolif. 36:131–149. 2003.
30	Lv XJ, Zhao LJ, Hao YQ, Su ZZ, Li JY, Du YW and Zhang J: Schisandrin B inhibits the proliferation of human lung adenocarcinoma A549 cells by inducing cycle arrest and apoptosis. Int J Clin Exp Med. 8:6926–6936. 2015.
31	Fushimi K, Ray P, Kar A, Wang L, Sutherland LC and Wu JY: Up-regulation of the proapoptotic caspase 2 splicing isoform by a candidate tumor suppressor, RBM5. Proc Natl Acad Sci USA. 105:15708–15713. 2008.
32	Rintala-Maki ND and Sutherland LC: LUCA-15/RBM5, a putative tumour suppressor, enhances multiple receptor-initiated death signals. Apoptosis. 9:475–484. 2004.
33	Rintala-Maki ND, Abrasonis V, Burd M and Sutherland LC: Genetic instability of RBM5/LUCA-15/H37 in MCF-7 breast carcinoma sublines may affect susceptibility to apoptosis. Cell Biochem Funct. 22:307–313. 2004.
34	Sutherland LC, Lerman M, Williams GT and Miller BA: LUCA-15 suppresses CD95-mediated apoptosis in Jurkat T cells. Oncogene. 20:2713–2719. 2001.
35	Sleeman J and Steeg PS: Cancer metastasis as a therapeutic target. Eur J Cancer. 46:1177–1180. 2010.
36	Verma S, Kesh K, Ganguly N, Jana S and Swarnakar S: Matrix metalloproteinases and gastrointestinal cancers: Impacts of dietary antioxidants. World J Biol Chem. 5:355–376. 2014.
37	Gencer S, Cebeci A and Irmak-Yazicioglu MB: Matrix metalloproteinase gene expressions might be oxidative stress targets in gastric cancer cell lines. Chin J Cancer Res. 25:322–333. 2013.
38	Ramakrishnan S, Subramanian IV, Yokoyama Y and Geller M: Angiogenesis in normal and neoplastic ovaries. Angiogenesis. 8:169–182. 2005.
39	Yamakuchi M, Lotterman CD, Bao C, Hruban RH, Karim B, Mendell JT, Huso D and Lowenstein CJ: P53-induced microRNA-107 inhibits HIF-1 and tumor angiogenesis. Proc Natl Acad Sci USA. 107:6334–6339. 2010.