ELMOD2对胃癌细胞MGC803恶性生物学行为的影响
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摘要
目的:探讨ELMOD2过表达对胃癌细胞MGC803恶性生物学行为的影响,并初步研究其相关的分子机制。方法:将GV141-ELMOD2表达载体转染人胃癌MGC803细胞,qPCR及WB实验检测MGC803细胞中ELMOD2 mRNA和蛋白的表达,CCK-8法检测MGC803细胞增殖能力,流式细胞术检测MGC803细胞凋亡情况,Transwell法检测MGC803细胞迁移能力,WB实验检测细胞中PCNA、BAX和Bcl-2以及Vimentin蛋白的表达。结果:ELMOD2表达载体转染后,MGC803细胞中ELMOD2mRNA和蛋白表达水平均显著提高(P<0.05)。ELMOD2过表达可使MGC803细胞增殖、迁移能力显著提高,凋亡率显著降低(P<0.05或P<0.01);细胞中PCNA、Vimentin、Bcl-2蛋白水平增高,BAX蛋白水平降低(P<0.05或P<0.01)。结论:ELMOD2过表达可促进MGC803细胞增殖、迁移,抑制细胞凋亡,上述效应可能部分通过提高PCNA、Vimentin、Bcl-2蛋白水平及降低BAX蛋白水平实现。
Objective: To investigate the effect of ELMOD2 over-expression on the malignant biological behaviors of gastric cancer MGC803 cells, and to study its related molecular mechanism. Methods: GV141-ELMOD2 expression vector was transfected into human gastric cancer MGC803 cells. The m RNA and protein expressions of ELMOD2 were detected by Real-time fluorescent quantitative PCR and WB, respectively. The cell proliferation ability was detected by CCK-8 method. Apoptosis rate was detected by flow cytometry. The cell migration ability was detected by Transwell method. The protein expressions of PCNA, BAX and Bcl-2 and Vimentin were detected by WB. Results: After transfection of ELMOD2 expression vector, the mRNA and protein expressions of ELMOD2 were significantly increased in MGC803 cells(P<0.05). Further studies showed that over-expression of ELMOD2 increased the proliferation and migration ability but reduced the apoptosis rate of MGC803 cells significantly(P<0.05 or P<0.01). The protein levels of PCNA,Vimentin and Bcl-2 in MGC803 cells increased, while the protein level of BAX decreased(P<0.05 or P<0.01). Conclusion: Over-expression of ELMOD2 can promote the proliferation and migration of MGC803 cells and inhibit cell apoptosis. These effects may be achieved by increasing the protein level of PCNA, Vimentin and Bcl-2, and reducing the protein level of BAX.
Objective: To investigate the effect of ELMOD2 over-expression on the malignant biological behaviors of gastric cancer MGC803 cells, and to study its related molecular mechanism. Methods: GV141-ELMOD2 expression vector was transfected into human gastric cancer MGC803 cells. The m RNA and protein expressions of ELMOD2 were detected by Real-time fluorescent quantitative PCR and WB, respectively. The cell proliferation ability was detected by CCK-8 method. Apoptosis rate was detected by flow cytometry. The cell migration ability was detected by Transwell method. The protein expressions of PCNA, BAX and Bcl-2 and Vimentin were detected by WB. Results: After transfection of ELMOD2 expression vector, the mRNA and protein expressions of ELMOD2 were significantly increased in MGC803 cells(P<0.05). Further studies showed that over-expression of ELMOD2 increased the proliferation and migration ability but reduced the apoptosis rate of MGC803 cells significantly(P<0.05 or P<0.01). The protein levels of PCNA,Vimentin and Bcl-2 in MGC803 cells increased, while the protein level of BAX decreased(P<0.05 or P<0.01). Conclusion: Over-expression of ELMOD2 can promote the proliferation and migration of MGC803 cells and inhibit cell apoptosis. These effects may be achieved by increasing the protein level of PCNA, Vimentin and Bcl-2, and reducing the protein level of BAX.
引文
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[23]ZHANG W T,ZHANG J F,ZHANG Z W,et al.Overexpression of indoleamine 2,3-dioxygenase 1 promotes epithelial-mesenchymal transition by activation of the IL-6/STAT3/PD-L1 pathway in bladder cancer[J/OL].Transl Oncol,2019,12(3):485-492[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6307990/.DOI:10.1016/j.tranon.2018.11.012.
[24]KOBAYASHI A,KUBOTA S,MORI N,et al.Photoreceptor synaptic protein HRG4(UNC119)interacts with Arl2 via a putative conserved domain[J].FEBS Lett,2003,534(1/2/3):26-32.
[25]LEI B,CHAI W,WANG Z Y,et al.Highly expressed UNC119 promotes hepatocellular carcinoma cell proliferation through Wnt/β-catenin signaling and predicts a poor prognosis[J/OL].Am J Cancer Res,2015,5(10):3123-3134[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4656735/.
[26]LIU Z H,ZHANG Y F,XU Z D.UNC119 promotes the growth and migration of hepatocellular carcinoma via Wnt/β-catenin and TGF-β/EMT signaling pathways[J].J BUON,2018,23(1):185-187.
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[28]PENG R Q,MEN J L,MA R,et al.MiR-214 down-regulates ARL2and suppresses growth and invasion of cervical cancer cells[J].Biochem Biophys Res Commun,2017,484(3):623-630.DOI:10.1016/j.bbrc.2017.01.152.
[29]LONG L M,HE B F,HUANG G Q,et al.MicroRNA-214 functions as a tumor suppressor in human colon cancer via the suppression of ADP-ribosylation factor-like protein 2[J/OL].Oncol Lett,2015,9(2):645-650[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4301474/.DOI:10.3892/ol.2014.2746.
[2]DANG S C,FAN Y Y,CUI L,et al.PLK1 as a potential prognostic marker of gastric cancer through MEK-ERK pathway on PDTXmodels[J/OL].Onco Targets Ther,2018,11:6239-6247[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6163028/.DOI:10.2147/OTT.S169880.
[3]JOHNSON K R,LONGO-GUESS C M,GAGNON L H.Mutations of the mouse ELMO domain containing 1 gene(Elmod1)link small GTPase signaling to actin cytoskeleton dynamics in hair cell stereocilia[J/OL].PLoS One,2012,7(4):e36074[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3338648/.DOI:10.1371/journal.pone.0036074.
[4]HODGSON U,PULKKINEN V,DIXON M,et al.ELMOD2 is a candidate gene for familial idiopathic pulmonary fibrosis[J/OL].Am J Hum Genet,2006,79(1):149-154[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1474126/.DOI:10.1086/504639.
[5]PULKKINEN V,BRUCE S,RINTAHAKA J,et al.ELMOD2,a candidate gene for idiopathic pulmonary fibrosis,regulates antiviral responses[J].FASEB J,2010,24(4):1167-1177.DOI:10.1096/fj.09-138545.
[6]SUZUKI M,MURAKAMI T,CHENG J L,et al.ELMOD2 is anchored to lipid droplets by palmitoylation and regulates adipocyte triglyceride lipase recruitment[J/OL].Mol Biol Cell,2015,26(12):2333-2342[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4462949/.DOI:10.1091/mbc.E14-11-1504.
[7]ZHOU C X,SHI L Y,LI R C,et al.GTPase-activating protein Elmod2 is essential for meiotic progression in mouse oocytes[J/OL].Cell Cycle,2017,16(9):852-860[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5444355/.DOI:10.1080/15384101.2017.1304329.
[8]GUO J F,BIAN Y,WANG Y,et al.S100A4 influences cancer stem cell-like properties of MGC803 gastric cancer cells by regulating GDF15 expression[J].Int J Oncol,2016,49(2):559-568.DOI:10.3892/ijo.2016.3556.
[9]HOU S S,TIAN T,QI D W,et al.S100A4 promotes lung tumor development throughβ-catenin pathway-mediated autophagy inhibition[J/OL].Cell Death Dis,2018,9(3):277[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5833421/.DOI:10.1038/s41419-018-0319-1.
[10]HUA J,CHEN D Q,FU H,et al.Short hairpin RNA-mediated inhibition of S100A4 promotes apoptosis and suppresses proliferation of BGC823 gastric cancer cells in vitro and in vivo[J].Cancer Lett,2010,292(1):41-47.DOI:10.1016/j.canlet.2009.11.007.
[11]RUMA I M W,KINOSHITA R,TOMONOBU N,et al.Embigin promotes prostate cancer progression by S100A4-dependent and-independent mechanisms[J/OL].Cancers(Basel),2018,10(7):E239[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6071117/.DOI:10.3390/cancers10070239.
[12]ZHANG J H,HOU S S,GU J C,et al.S100A4 promotes colon inflammation and colitis-associated colon tumorigenesis[J/OL].Oncoimmunology,2018,7(8):e1461301[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6136879/.DOI:10.1080/2162402X.2018.1461301.
[13]BANTIS A,GIANNOPOULOS A,GONIDI M,et al.Expression of p120,Ki-67 and PCNA as proliferation biomarkers in imprint smears of prostate carcinoma and their prognostic value[J].Cytopathology,2004,15(1):25-31.
[14]MATHEWS M B,BERNSTEIN R M,FRANZA B R Jr,et al.Identity of the proliferating cell nuclear antigen and cyclin[J].Nature,1984,309(5966):374-376.
[15]LI S R,SHI D,ZHANG L Y,et al.Oridonin enhances the radiosensitivity of lung cancer cells by upregulating BAX and downregulating Bcl-2[J/OL].Exp Ther Med,2018,16(6):4859-4864[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6256844/.DOI:10.3892/etm.2018.6803.
[16]ROSHANRAVAN N,ASGHARIAN P,DARIUSHNEJAD H,et al.eryngium Billardieri induces apoptosis via bax gene expression in pancreatic cancer cells[J/OL].Adv Pharm Bull,2018,8(4):667-674[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6311640/.DOI:10.15171/apb.2018.075.
[17]BATTAGLIA R A,DELIC S,HERRMANN H,et al.Vimentin on the move:new developments in cell migration[J/OL].F1000Res,2018,7:F1000 Faculty Rev-F1000 Faculty1796[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6241562/.DOI:10.12688/f1000research.15967.1.
[18]VUORILUOTO K,HAUGEN H,KIVILUOTO S,et al.Vimentin regulates EMT induction by Slug and oncogenic H-Ras and migration by governing Axl expression in breast cancer[J].Oncogene,2011,30(12):1436-1448.DOI:10.1038/onc.2010.509.
[19]MATOZAKI T,NAKANISHI H,TAKAI Y.Small G-protein networks:their crosstalk and signal cascades[J].Cell Signal,2000,12(8):515-524.
[20]MUROMOTO R,SEKINE Y,IMOTO S,et al.BART is essential for nuclear retention of STAT3[J].Int Immunol,2008,20(3):395-403.DOI:10.1093/intimm/dxm154.
[21]ZHANG T L,LI S,ZHANG Y C,et al.Crystal structure of the ARL2-GTP-BART complex reveals a novel recognition and binding mode of small GTPase with effector[J].Structure,2009,17(4):602-610.DOI:10.1016/j.str.2009.01.014.
[22]HU F Q,SUN X L,LI G,et al.Inhibition of SIRT2 limits tumour angiogenesis via inactivation of the STAT3/VEGFA signalling pathway[J/OL].Cell Death Dis,2018,10(1):9[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6315023/.DOI:10.1038/s41419-018-1260-z.
[23]ZHANG W T,ZHANG J F,ZHANG Z W,et al.Overexpression of indoleamine 2,3-dioxygenase 1 promotes epithelial-mesenchymal transition by activation of the IL-6/STAT3/PD-L1 pathway in bladder cancer[J/OL].Transl Oncol,2019,12(3):485-492[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6307990/.DOI:10.1016/j.tranon.2018.11.012.
[24]KOBAYASHI A,KUBOTA S,MORI N,et al.Photoreceptor synaptic protein HRG4(UNC119)interacts with Arl2 via a putative conserved domain[J].FEBS Lett,2003,534(1/2/3):26-32.
[25]LEI B,CHAI W,WANG Z Y,et al.Highly expressed UNC119 promotes hepatocellular carcinoma cell proliferation through Wnt/β-catenin signaling and predicts a poor prognosis[J/OL].Am J Cancer Res,2015,5(10):3123-3134[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4656735/.
[26]LIU Z H,ZHANG Y F,XU Z D.UNC119 promotes the growth and migration of hepatocellular carcinoma via Wnt/β-catenin and TGF-β/EMT signaling pathways[J].J BUON,2018,23(1):185-187.
[27]IWASA H,SARKAR A,SHIMIZU T,et al.UNC119 is a binding partner of tumor suppressor Ras-association domain family 6 and induces apoptosis and cell cycle arrest by MDM2 and p53[J/OL].Cancer Sci,2018,109(9):2767-2780[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6125449/.DOI:10.1111/cas.13706.
[28]PENG R Q,MEN J L,MA R,et al.MiR-214 down-regulates ARL2and suppresses growth and invasion of cervical cancer cells[J].Biochem Biophys Res Commun,2017,484(3):623-630.DOI:10.1016/j.bbrc.2017.01.152.
[29]LONG L M,HE B F,HUANG G Q,et al.MicroRNA-214 functions as a tumor suppressor in human colon cancer via the suppression of ADP-ribosylation factor-like protein 2[J/OL].Oncol Lett,2015,9(2):645-650[2019-02-22].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4301474/.DOI:10.3892/ol.2014.2746.