IKBKE和NF-κB在胰腺癌中的表达及沉默IKBKE对胰腺癌细胞增殖、迁移的影响

Expression of IKBKE and NF-κB in pancreatic cancer and the effect of IKBKE on proliferation and migration of pancreatic cancer cells

目的检测胰腺癌患者IKBKE和NF-κB的表达,并探索IKBKE对胰腺癌细胞的影响。方法免疫组织化学法检测天津医科大学第二医院2012年1月至2017年1月收治的61例患者的胰腺癌组织及13例正常胰腺组织IKBKE和NF-κB的表达情况,并分析其与患者临床病理特征的关系。用慢病毒介导的IKBKE RNAi转染胰腺癌细胞,从而敲低IKBKE的表达水平。采用蛋白印迹法检测沉默效率;CCK-8、平板克隆和划痕实验检测细胞的增殖、迁移能力。结果组织病理显示,胰腺癌组织中IKBKE染色呈弱阳性、阳性、强阳性者共有60(98.4%)例,高于正常胰腺组织(76.9%为弱阳性,其余阴性),差异有统计学意义(P<0.05)。NF-κB染色全部为弱阳性及以上,高于正常胰腺组织(30.8%为弱阳性,其余阴性),差异有统计学意义(P<0.05)。生存分析显示,IKBKE高表达患者的总生存期低于低表达患者,差异有统计学意义(P<0.05)。CCK-8、平板克隆和细胞划痕实验均表明,IKBKE敲低后细胞增殖和迁移能力低于对照组(均P<0.05)。结论IKBKE和NF-κB在胰腺癌中表达上调,且两者表达强度具有显著相关性。沉默IKBKE表达能明显抑制胰腺癌细胞的增殖和迁移。

Objective To investigate the expression of IKBKE and NF-κB in pancreatic cancer,and to explore the effect of IKBKE on pancreatic cancer proliferation and migration.Methods Immunohistochemistry staining was used to study the expression of IKBKE and NF-κB in tissues of 61 pancreatic cancer patients admitted to the Second Hospital of Tianjin Medical University from January 2012 to January 2017 and 13 normal pancreatic tissues.The correlations between those expression to clinicopathological features were analyzed.Lentivirus mediated RNAi was transfected into pancreatic cancer cells to block IKBKE.Western blot was performed to test the silencing effeciency;CCK-8 and plate clone and scratch assays were used to investigate the proliferation and migration of pancreatic cancer.Results Immunohistochemical staining showed that 60(98.4%)of IKBKE staining were weakly positive,positive,and strongly positive in pancreatic cancer tissues,which were significantly higher than normal pancreatic tissues(76.9%cases were weakly positive and the rest were negative),and the differences were statistically significant(P<0.05).All cases of NF-κB exhibited weakly positive expression and above in pancreatic cancer tissues,which was markedly higher than normal tissues(30.8%cases were weak positive and the rest were negative staining),statistically significant(P<0.05).Survival analysis showed that patients with high level of IKBKE showed a shorter overall survival(P<0.05).CCK-8,plate cloning and scratch assays showed that the proliferation and migration of were significantly decreased in IKBKE knocking down group(P<0.05).Conclusions IKBKE and NF-κB are highly expressed in pancreatic cancer,and IKBKE is correlated with NF-κB in pancreatic cancer.Blocking of IKBKE could distinctly inhibit the proliferation and migration of pancreatic cancer.