siRNA沉默ADAM17基因对食管癌EC109细胞增殖的影响
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摘要
目的探讨小干扰RNA(siRNA)对EC109食管癌细胞增殖的影响。方法采用RNA干扰技术靶向人ADAM17基因的siRNA,分别标记为si257、si258及si259 3个序列,转染入EC109人食管癌细胞;采用实时定量PCR法检测各组癌细胞ADAM17 mRNA的含量;采用CCK8实验检测ADAM17沉默后食管癌细胞EC109的细胞增殖变化。结果 3个序列均能够有效抑制EC109中ADAM17的表达,显示有效的基因沉默效果。转染后72 h si259对EC109的增殖能力具有抑制作用(P<0.05)。未观察到si257对EC109增殖的抑制作用。结论 siRNA沉默ADAM17基因能够对食管癌细胞增殖产生一定的影响,提示ADMA17基因通过某种通道参与了食管癌细胞的增殖活动。
Objective To investigate the effect of small interfering RNA(siRNA) on the proliferation of EC109 esophageal carcinoma cells.Methods The small siRNA targeting human ADAM17 gene by RNA interference technique was labeled as si257,si258 and si259 sequences,which were transfected into EC109 cells.The ADAM17 mRNA expression of each group was detected by quantitative real time-PCR assay.The CCK8 assay was used to detect the proliferation of EC109 cells line after ADAM17 silencing.Results Three sequences could effectively inhibit the expression of ADAM17 in EC109 cells and showed effective gene silencing effect.Si259 could significantly inhibit the proliferation of EC109 on the 72~(nd) hour after interference.The inhibitory effect of si257 on the proliferation of EC109 was not observed.Conclusion siRNA targeting ADAM17 gene may have a certain effect on the proliferation of esophageal carcinoma cells,suggesting that ADMA17 gene plays an important role in the proliferation of esophageal carcinoma cells through a certain channel.
Objective To investigate the effect of small interfering RNA(siRNA) on the proliferation of EC109 esophageal carcinoma cells.Methods The small siRNA targeting human ADAM17 gene by RNA interference technique was labeled as si257,si258 and si259 sequences,which were transfected into EC109 cells.The ADAM17 mRNA expression of each group was detected by quantitative real time-PCR assay.The CCK8 assay was used to detect the proliferation of EC109 cells line after ADAM17 silencing.Results Three sequences could effectively inhibit the expression of ADAM17 in EC109 cells and showed effective gene silencing effect.Si259 could significantly inhibit the proliferation of EC109 on the 72~(nd) hour after interference.The inhibitory effect of si257 on the proliferation of EC109 was not observed.Conclusion siRNA targeting ADAM17 gene may have a certain effect on the proliferation of esophageal carcinoma cells,suggesting that ADMA17 gene plays an important role in the proliferation of esophageal carcinoma cells through a certain channel.
引文
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[12] 刘宏斌,朱燕,杨其昌,等.ADAM17蛋白在食管鳞癌组织中表达及其临床病理意义[J].江苏医药,2012,38(10):1180-1182.
[2] Zhang Q,Wang C,Han X,et al.Knockdown of ADAM17 inhibits cell proliferation and increases oxaliplatin sensitivity in HCT-8 colorectal cancer through EGFR-PI3K-AKT activation[J].Biochemical and Biophysical Research Communications,2018,503(4):2333-2339.
[3] Le Gall SM,Bobe P,Reiss K,et al.ADAMs 10 and 17 represent differentially regulated components of a general shedding machinery for membrane proteins such as transforming growth factor alpha,L-selectin,and tumor necrosis factor alpha[J].Mol Biol Cell,2009,20:1785-1794.
[4] Saftig P,Reiss K.The “A Disintegrin And Metalloproteases” ADAM10 and ADAM17:novel drug targets with therapeutic potential[J]?Eur J Cell Biol,2011,90(6-7):527-535.
[5] Lorenzen I,Trad A,Grozinger J.Multimerisation of a disintegrin and metalloprotease protein-17 (ADAM17) is mediated by its EGF-like domain[J].Biochem Biophys Res Commun,2011,415(2):330-336.
[6] McGowan PM,Ryan BM,Hill AD,et al.ADAM-17 expression in breast cancer correlates with variables of tumor progression[J].Clin Cancer Res,2007,13(8):2335-2343.
[7] Sakamoto N,Oue N,Noguchi T,et al.Serial analysis of gene expression of esophageal squamous cell carcinoma:ADAMTS16 is upregulated in esophageal squamous cell carcinoma[J].Cancer Sci,2010,101:1038-1044.
[8] Liu HB,Yang QC,Sheng YI,et al.A disintegrin and metalloproteinase 17 mRNA and protein expression in esophageal squamous cell carcinoma,as well as its clinicopathological factors and prognosis[J].Molecular Med Rep,2015,11(2):961-967.
[9] Liu HB,Zhu Y,Yang QC,et al.Expression and clinical significance of ADAMA17 protein in esophageal squamous cell carcinoma[J].Genetics and Molecular Research,2015,14(2):4391-4398.
[10] 刘宏斌,杨其昌,沈屹,等.去整合素-金属蛋白酶17 mRNA和蛋白在食管鳞癌组织中的表达及其临床意义[J].中华肿瘤杂志,2013,35(5):361-365.
[11] 刘宏斌,杨其昌,沈屹,等.食管鳞状细胞癌组织中去整合素-金属蛋白酶17mRNA表达的临床病理意义[J].肿瘤研究与临床,2012,24(7):444-447.
[12] 刘宏斌,朱燕,杨其昌,等.ADAM17蛋白在食管鳞癌组织中表达及其临床病理意义[J].江苏医药,2012,38(10):1180-1182.