下调miR-92a对非小细胞肺癌细胞增殖及血管生成因子表达的影响
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摘要
目的探讨miR-92a下调对非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞增殖及血管生成的影响。方法将NSCLC中A549细胞分为三组培养:A549组(未转染A549细胞)、sc-siRNA组(转染sc-siRNA的A549细胞)、miR-92a-siRNA组(转染miR-92a-siRNA的A549细胞)。分别采用RT-PCR和Western blot检测A549细胞和正常人支气管上皮(human bronchial epithelial,HBE)细胞中miR-92a相对表达量、PTEN、血管内皮生长因子(vascular endothelial growth factor,VEGF)蛋白相对表达水平。采用活细胞计数法和结晶紫染色实验检测各组A549细胞的增殖能力。结果 miR-92a在A549组细胞中的相对表达量高于HBE细胞(P <0. 05); A549组细胞中PTEN蛋白表达水平低于HBE细胞(P <0. 05),VEGF蛋白表达水平高于HBE细胞(P <0. 05); miR-92a-siRNA组细胞中miR-92a相对表达量降低(P <0. 05),PTEN蛋白表达水平升高(P <0. 05),VEGF蛋白表达水平降低(P <0. 05); miR-92a-siRNA组细胞中PI3K和Akt蛋白表达水平均下降(P <0. 05); miR-92a-siRNA组细胞数目减少,细胞增殖能力减弱。结论 NSCLC中A549细胞的miR-92a表达明显上调,miR-92a基因沉默能明显抑制细胞增殖、血管生成,PTEN和VEGF相关PI3K/Akt信号通路可能在此过程中发挥重要作用。
Purpose To investigate the effect of down-regulation of miR-92 a on the proliferation and angiogenesis of nonsmall cell lung cancer( NSCLC). Methods Human NSCLC cell A549 was divided into three groups: A549 group( non-transfected A549 cells),sc-siRNA group( A549 cells transfected with sc-siRNA) and miR-92a-siRNA group( A 5 4 9 cells trans-fected with miR-92a-siRNA). The relative expression level of miR-92 a,PTEN and vascular endothelial growth factor( VEGF)in A549 cells and human bronchial epithelial( HBE) cells were detected by RT-PCR and Western blot respectively. The proliferation ability of A549 cells in each group was detected by living cell count and crystal violet staining experiment. Results The relative expression of miR-92 a in A549 cells was significantly higher than that in HBE cells( P < 0. 05),the expression level of PTEN protein in A549 cells was significantly lower than that in HBE cells( P < 0. 05),and the expression level of VEGF protein was significantly higher than that in HBE cells( P < 0. 05).In the miR-92a-siRNA group,the relative expression of miR-92 a decreased( P < 0. 05),the expression level of PTEN protein in-creased( P < 0. 05),and the expression level of VEGF protein decreased( P < 0. 05). The expression levels of PI3 K and Akt in miR-92a-siRNA group decreased( P < 0. 05). the number of cells and cell proliferation ability in miR-92a-siRNA group reduced. Conclusion The expression of miR-92 a in NSCLC A549 cells is up-regulated,miR-92 a gene silencing can significantly inhibit cell proliferation and inhibit cell angiogenesis,PTEN and VEGF related PI3K/Akt signaling pathways may play an important role in this process.
Purpose To investigate the effect of down-regulation of miR-92 a on the proliferation and angiogenesis of nonsmall cell lung cancer( NSCLC). Methods Human NSCLC cell A549 was divided into three groups: A549 group( non-transfected A549 cells),sc-siRNA group( A549 cells transfected with sc-siRNA) and miR-92a-siRNA group( A 5 4 9 cells trans-fected with miR-92a-siRNA). The relative expression level of miR-92 a,PTEN and vascular endothelial growth factor( VEGF)in A549 cells and human bronchial epithelial( HBE) cells were detected by RT-PCR and Western blot respectively. The proliferation ability of A549 cells in each group was detected by living cell count and crystal violet staining experiment. Results The relative expression of miR-92 a in A549 cells was significantly higher than that in HBE cells( P < 0. 05),the expression level of PTEN protein in A549 cells was significantly lower than that in HBE cells( P < 0. 05),and the expression level of VEGF protein was significantly higher than that in HBE cells( P < 0. 05).In the miR-92a-siRNA group,the relative expression of miR-92 a decreased( P < 0. 05),the expression level of PTEN protein in-creased( P < 0. 05),and the expression level of VEGF protein decreased( P < 0. 05). The expression levels of PI3 K and Akt in miR-92a-siRNA group decreased( P < 0. 05). the number of cells and cell proliferation ability in miR-92a-siRNA group reduced. Conclusion The expression of miR-92 a in NSCLC A549 cells is up-regulated,miR-92 a gene silencing can significantly inhibit cell proliferation and inhibit cell angiogenesis,PTEN and VEGF related PI3K/Akt signaling pathways may play an important role in this process.
引文
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[13]Ren P,Gong F,Zhang Y,et al.MicroRNA-92a promotes growth,metastasis,and chemoresistance in non-small cell lung cancer cells by targeting PTEN[J].Tumor Biol,2016,37(3):3215-3225.
[14]王秀,杜冀晖,黄虞,等.miR-92a在结直肠癌中的表达及其对肿瘤血管生成的作用[J].中国肿瘤临床,2016,43(6):223-227.
[15]Lu X X,Cao L Y,Chen X,et al.PTEN inhibits cell proliferation,promotes cell apoptosis,and induces cell cycle arrest via downregulating the PI3K/AKT/hTERT pathway in lung adenocarcinoma A549 cells[J].Biomed Res Int,2016,2016(12):2476842.
[2]Wei S,Tian J,Song X,et al.Causes of death and competing risk analysis of the associated factors for non-small cell lung cancer using the surveillance,epidemiology,and end results database[J].J Cancer Res Clin Oncol,2018,144(1):145-155.
[3]Elshafei A,Shaker O,Abd E O,et al.The expression profiling of serum miR-92a,miR-375,and miR-760 in colorectal cancer:an egyptian study[J].Tumour Biol,2017,39(6):1-14.
[4]Gong J,He L,Ma J,et al.The relationship between miR-17-5p,miR-92a,and let-7b expression with non-small cell lung cancer targeted drug resistance[J].J BUON,2017,22(2):454-461.
[5]Xu X,Zhu S,Tao Z,et al.High circulating miR-18a,miR-20a,and miR-92a expression correlates with poor prognosis in patients with non-small cell lung cancer[J].Cancer Med,2018,7(1):21-31.
[6]唐凤娟,许三鹏,王国平,等.Wnt5a和Wnt11在肺癌中的表达及意义[J].临床与实验病理学杂志,2017,33(10):1082-1086.
[7]许晓阳,吴淑华,孙晨博,等.结直肠癌中PHLPP2、PTEN与p-Akt1表达的相关性及其临床意义[J].临床与实验病理学杂志,2018,34(8):827-833.
[8]Zhu X,Li Z,Li T,et al.Osthole inhibits the PI3K/AKT signaling pathway via activation of PTEN and induces cell cycle arrest and apoptosis in esophageal squamous cell carcinoma[J].Biomed Pharmacother,2018,102:502-509.
[9]赵军,皇甫辉.PI3K/Akt信号通路及血管内皮生长因子对肿瘤血管生成的影响[J].国际耳鼻咽喉头颈外科杂志,2014,38(1):5-6.
[10]Smith L,Baxter E W,Chambers P A,et al.Down-regulation of miR-92 in breast epithelial cells and in normal but not tumour fibroblasts contributes to breast carcinogenesis[J].PLo S One,2015,10(10):1-17.
[11]Zhou C,Shen L,Mao L,et al.miR-92a is upregulated in cervical cancer and promotes cell proliferation and invasion by targeting FBXW7[J].Biochem Biophys Res Commun,2015,458(1):63-69.
[12]Zhang G,Zhou H,Xiao H,et al.MicroRNA-92a functions as an oncogene in colorectal cancer by targeting PTEN[J].Dig Dis Sci,2014,59(1):98-107.
[13]Ren P,Gong F,Zhang Y,et al.MicroRNA-92a promotes growth,metastasis,and chemoresistance in non-small cell lung cancer cells by targeting PTEN[J].Tumor Biol,2016,37(3):3215-3225.
[14]王秀,杜冀晖,黄虞,等.miR-92a在结直肠癌中的表达及其对肿瘤血管生成的作用[J].中国肿瘤临床,2016,43(6):223-227.
[15]Lu X X,Cao L Y,Chen X,et al.PTEN inhibits cell proliferation,promotes cell apoptosis,and induces cell cycle arrest via downregulating the PI3K/AKT/hTERT pathway in lung adenocarcinoma A549 cells[J].Biomed Res Int,2016,2016(12):2476842.