过表达CASK基因对非小细胞肺癌H1299细胞凋亡及周期的影响
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摘要
目的:探讨过表达人钙/钙调蛋白依赖性丝氨酸蛋白激酶(CASK)基因对非小细胞肺癌(NSCLC)H1299细胞凋亡及周期的影响。方法:将本课题组前期构建的CASK基因过表达慢病毒感染经体外培养的NSCLC H1299细胞(OE组),同时以CON328阴性对照病毒感染H1299细胞(NC组),并设空白对照组(MOCK组)。流式细胞术检测各组细胞的凋亡情况及细胞周期变化。结果:OE组与NC组及MOCK组比较,细胞凋亡率明显升高(P<0.05)。而NC组凋亡率与MOCK组比较,差异无统计学意义(P>0.05);与NC组和MOCK组比较,OE组G1期细胞数显著增多,而S期和G2/M期细胞数显著减少(P<0.05)。结论:CASK基因过表达可以显著升高NSCLC H1299细胞凋亡率,细胞周期停滞,细胞增殖缓慢,进而抑制肺癌的发展。
Objective:To investigate the effect of overexpression of calcium/calmodulin dependent serine protein kinase(CASK) gene on the apoptosis and cell cycle of non-small cell lung cancer(NSCLC) H1299 cells.Methods:H1299 cells transfected with lentiviral vector over-expressing CASK(OE group) or negative control virus particles(NC group),and uninfected H1299 cells served as the MOCK group.Flow cytometry was used to detect cell apoptosis and cell cycle.Results:The apoptotic rate in the OE group was significantly higher than that in the NC and MOCK groups(P<0.05),and no obvious difference was found in NC group and MOCK group(P>0.05).OE group had more cells in G1 stage and fewer cells in S and G2/M stages,compared with NC group and MOCK group(P<0.05).Conclusion:Overexpression of CASK could increase NSCLC H1299 cells apoptosis,inhibit cell proliferation and the development of lung cancer.
Objective:To investigate the effect of overexpression of calcium/calmodulin dependent serine protein kinase(CASK) gene on the apoptosis and cell cycle of non-small cell lung cancer(NSCLC) H1299 cells.Methods:H1299 cells transfected with lentiviral vector over-expressing CASK(OE group) or negative control virus particles(NC group),and uninfected H1299 cells served as the MOCK group.Flow cytometry was used to detect cell apoptosis and cell cycle.Results:The apoptotic rate in the OE group was significantly higher than that in the NC and MOCK groups(P<0.05),and no obvious difference was found in NC group and MOCK group(P>0.05).OE group had more cells in G1 stage and fewer cells in S and G2/M stages,compared with NC group and MOCK group(P<0.05).Conclusion:Overexpression of CASK could increase NSCLC H1299 cells apoptosis,inhibit cell proliferation and the development of lung cancer.
引文
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[4]WEI J L,FU Z X,FANG M,et al.High expression of CASK correlates with progression and poor prognosis of colorectal cancer[J].Tumour Biol,2014,35(9):9185-9194.
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[6]MOK T S,WU Y L,AHN M J,et al.Osimertinib or platinum-pemetrexed in EGFR T790M-positive lung cancer[J].New England Journal of Medicine,2017,376(7):629-640.
[7]LADANYI M,PAO W.Lung adenocarcinoma:guiding EGFR-targeted therapy and beyond[J].Modern Pathology,2008,21(S2):S16.
[8]CHEN Z,SASAKI T,TAN X,et al.Inhibition of ALK,PI3K/MEK,and HSP90in murine lung adenocarcinoma induced by EML4-ALK fusion oncogene[J].Cancer Research,2010,70(23):9827-9836.
[9]RIZVI N A,HELLMANN M D,SNYDER A,et al.Mutational landscape determines sensitivity to PD-1blockade in non-small cell lung cancer[J].Science,2015,348(6230):124-128.
[10]COHEN A R,WOODS D F,MARFATIA S M,et al.Human CASK/LIN-2binds syndecan-2and protein 4.1and localizes to the basolateral membrane of epithelial cells[J].J Cell Biol,1998,142(1):129-138.
[11]WEIGAND J E,JES-NIELS B,PASCAL G,et al.Hypoxia-induced alternative splicing in endothelial cells[J].PLoS One,2012,7(8):e42697.
[12]CARUANA G.Genetic studies define MAGUK proteins as regulators of epithelial cell polarity[J].Int J Dev Biol,2002,46(4):511-518.
[13]HSUEH Y P,WANG T F,YANG F C,et al.Nuclear translocation and transcription regulation by the membrane-associated guanylate kinase CASK/LIN-2[J].Nature,2000,404(6775):298-302.
[14]WARAYA M,YAMASHITA K,EMA A,et al.Exclusive association of p53 mutation with super-high methylation of tumor suppressor genes in the p53pathway in a unique gastric cancer phenotype[J].PLoS One,2015,10(10):e0139902.
[15]HUANG T N,CHANG H P,HSUEH Y P.CASKphosphorylation by PKA regulates the protein-protein interactions of CASK and expression of the NMDAR2b gene[J].Journal of Neurochemistry,2010,112(6):1562-1573.
[16]GIANSANTI F,SABATINI D,PENNACCHIO M R,et al.PDZ domain in the engineering and production of a saporin chimeric toxin as a tool for targeting cancer cells[J].Journal of Cellular Biochemistry,2015,116(7):1256-1266.
[17]DAVIS H M,PACHECO-COSTA R,ATKINSONE G,et al.Disruption of the Cx43/miR21pathway leads to osteocyte apoptosis and increased osteoclastogenesis with aging[J].Aging Cell,2017,16(3):551-563.
[18]RONGJU S,YONGYUE S,XIAODONG Z,et al.Human calcium/calmodulin-dependent serine protein kinase regulates the expression of p21via the E2Atranscription factor[J].Biochemical Journal,2009,419(2):457-466.
[19]HYO-SOO K,HYUN-JAI C,HYUN-JU C,et al.The essential role of p21in radiation-induced cell cycle arrest of vascular smooth muscle cell[J].Journal of Molecular&Cellular Cardiology,2004,37(4):871-880.