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胰腺癌小鼠模型中核仁蛋白14和CD31表达与肿瘤进展的相关性研究

Expression of nucleolar protein 14 and CD31 in pancreatic cancer mouse model and its correlation with tumor progression

摘要:

目的 探讨胰腺癌小鼠模型中核仁蛋白14(NOP14)和CD31的表达水平与肿瘤进展的相关性.方法 收集2013年1月至2015年12月经北京协和医院病理学检查证实伴肝转移的5例胰腺导管腺癌患者的临床病理资料,通过免疫组化方法检测胰腺癌原发肿瘤和相应肝转移灶中NOP14的表达水平;利用慢病毒表达载体感染胰腺癌细胞株,构建NOP14稳定下调的细胞株,实时荧光定量PCR和免疫印迹法检测NOP14抑制率;利用胰腺癌小鼠模型分析NOP14敲低组(n=8)和对照组(n=8)肿瘤组织中CD31标记的肿瘤微血管密度(MVD)及其与肿瘤大小和转移的关系.采用Mann Whitney U检验对组间数据进行统计学分析,应用Pearson 或Spearman相关系数分析变量间的相关性.结果 NOP14在胰腺癌肝转移灶中的表达水平较原发肿瘤明显升高(2.09±0.45比1.31±0.27,P=0.028);实时荧光定量PCR和免疫印迹法证实胰腺癌稳定转染细胞株中NOP14的表达水平分别下调了86%和78%;胰腺癌小鼠皮下和原位移植模型中NOP14敲低组MVD值(61.40±13.85和38.33±10.91)低于对照组(85.53±14.59和59.33±15.37)(P =0.041、P =0.037),且与肿瘤大小(r=0.842,P<0.01)和转移(r=0.726,P=0.008)呈正相关.结论 NOP14在胰腺癌肝转移灶中表达上调,可能通过促进肿瘤微血管形成导致胰腺癌的生长和转移.

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abstracts:

Objective To investigate expression of nucleolar protein 14(NOP14) and CD31 in pancreatic cancer mouse model and its correlation with tumor progression.Methods Clinicopathological data of 5 patients with pathologically confirmed pancreatic ductal adenocarcinoma(PDAC) and hepatic metastasis between January 2013 and December 2015 was collected in Department of General Surgery,Peking Union Medical College Hospital.Immunohistochemistry staining was employed to detect the expression of NOP14 in matched primary PDAC and relevant metastasis.Pancreatic cancer cells with NOP14 stably knocked down were established by transfecting lentivirus with NOP14 targeted silencing RNA.The inhibition efficacy was detected by quantitative real time PCR and western blot.Microvascular density(MVD) in pancreatic cancer transplantation mouse model was determined by CD31 immunohistochemistry staining analysis and correlated with NOP14 expression and tumor progression.Results NOP14 had a significant higher expression in liver metastasis than primary pancreatic adenocarcinoma (2.09±0.45 vs.1.31±0.27,P=0.028).NOP14 was knocked down 86 percent on mRNA level determined by qPCR and 78 percents on protein level detected by western blot.MVD was significantly decreased in NOP14-inhibited tumor from both pancreatic cancer cells subcutaneously and orthotopically grafted tumor mouse model with the value of 61.40±13.85 vs.85.53±14.59 (P=0.041) and 38.33±10.91 vs.59.33±15.37(P =0.037),respectively.Besides, MVD was positively associated with tumor volume(r=0.842,P<0.01) and metastasis (r=0.726, P=0.008).Conclusion NOP14 presents higher expression in hepatic metastasis of pancreatic adenocarcinoma and might promote tumor progression by increasing microvascular density.

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作者: 杜永星 [1] 李宗泽 [1] 刘长征 [2] 刘子文 [1]
第一作者: 杜永星
期刊: 《中华外科杂志》2017年55卷6期 463-467页 MEDLINEISTICPKUCSCD
栏目名称: 论著
DOI: 10.3760/cma.j.issn.0529-5815.2017.06.013
发布时间: 2017-06-30
基金项目:
国家自然科学基金面上项目 公益性行业科研专项(201402001)National Natural Science Foundation of China (General Program) Special Scientific Research Fund of Public Welfare Profession of China
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