摘要
目的:观察干扰前蛋白转化酶枯草溶菌素9(PCSK9)表达对胆管癌细胞增殖和迁移的影响并探讨可能的机制。方法:实时荧光定量PCR(RT-qPCR)检测PCSK9mRNA在胆管癌细胞TFK-1、HCCC-9810、HuCC-T1、QBC939、RBE和永生化胆管上皮细胞HIBEC中的表达。根据不同处理方法将HuCC-T1细胞分为si-PCSK9组和si-NC组。RT-qPCR和Westernblot检测各组HuCC-T1细胞中PCSK9的表达水平。CCK-8实验和划痕实验分别检测HuCC-T1细胞增殖活性和迁移能力。Westernblot检测核因子-κB(NF-κB)信号通路相关蛋白表达。结果:HCC-1937、MCF-7、BT-549、HuCC-T1细胞PCSK9mRNA表达明显高于HIBEC细胞,且HuCC-T1细胞PCSK9 mRNA表达水平最高(均P<0.01)。si-PCSK9组HuCC-T1细胞中PCSK9表达明显低于si-NC组(P<0.01)。干扰PCSK9表达后,HuCC-T1细胞的增殖活性降低(P<0.05)。si-PCSK9组划痕愈合率低于si-NC组(P<0.01)。与si-NC组比较,si-PCSK9组HuCC-T1细胞中NF-κB信号通路蛋白高尔基体外周膜蛋白P65(p65)、粒细胞集落刺激因子(G-CSF)、基质金属蛋白酶-9(MMP-9)、血管内皮生长因子(VEGF)表达明显降低(均P<0.01)。结论:胆管癌细胞中PCSK9呈高表达,干扰PCSK9可能通过降低NF-κB信号通路活性抑制胆管癌HuCC-T1细胞的增殖和迁移。
Objective:To observe the effect of the interference of proprotein convertase subtilisin/kexin type 9(PCSK9)expression on the proliferation and migration of cholangiocarcinoma cells and explore the possible mechanism.Methods:RT-qPCRwas used to detect PCSK9 mRNA expression in cholangiocarcinoma cells TFK-1,HCCC-9810,HuCC-T1,QBC939,RBE and immortalized cholangioepithelial cells HIBEC.HuCC-T1 cells were divided into si-PCSK9 group and si-NC group according to different treatment methods.RT-qPCRand Western blot were used to detect PCSK9 expression in each group of HuCC-T1 cells.CCK-8 experiment and scratch experiment were used to detect the proliferation activity and migration ability of HuCC-T1 cells.Western blot was used to detect NF-κB signaling pathway proteins expression in HuCC-T1 cells.Results:Compared with HIBEC cells,PCSK9 mRNA was higher in cholangiocarcinoma cells TFK-1,HCCC-9810,HuCC-T1,QBC939,and RBE,and PCSK9 mRNA expression was highest in HuCC-T1 cells(all P<0.01).The expression of PCSK9 in HuCC-T1 cells in si-PCSK9 group was significantly lower than that in si-NC group(P<0.01).After interfering with PCSK9 expression,the proliferation activity of HuCC-T1 cells was reduced(P<0.05).The scratch healing rate of si-PCSK9 group was lower than that of si-NC group(P<0.01).Compared with the si-NC group,the expressions of NF-κB signaling pathway proteins p65,G-CSF,MMP-9,and VEGF in HuCC-T1 cells were significantly reduced(all P<0.01).Conclusion:PCSK9 is highly expressed in cholangiocarcinoma cells,and interference with PCSK9 may inhibit the proliferation and migration of HuCC-T1 cells by decreasing the activity of NF-κB signaling pathway.
作者
崔龙泉
徐红伟
茹怡婷
何辉
CUI Longquan;XU Hongwei;RU Yiting;HE Hui(Department of Hepatobiliary Surgery,the First Affiliated Hospital of Xinxiang Medical College,Xinxiang 453100,China;General Practice of Traditional Chinese Medicine,Lingyun Community Health Service Center,Xuhui District,Shanghai 200237,China;Department of General Surgery,the First Affiliated Hospital of Dalian Medical University,Dalian 116021,China)
出处
《陕西医学杂志》
2025年第3期291-295,共5页
Shaanxi Medical Journal
基金
国家自然科学基金资助项目(81903964)。
