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siRNA干扰Gab1表达对胆管癌细胞株HUCCA-1功能的影响及其与PI3K/Akt信号通路的关系 被引量:1
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作者 姚旭 田忠 刘源 《天津医药》 CAS 2015年第6期590-594,I0003,共6页
目的:检测下调Gab1表达后人胆管癌细胞株HUCCA-1细胞功能、PI3KCA及Akt1的蛋白及mRNA表达水平改变,探讨Gab1、PI3KCA及Akt1表达在胆管癌恶性行为中的作用机制。方法 Gab1siRNA转染HUCCA-1细胞,采用qRT-PCR法及Western blot法检测转... 目的:检测下调Gab1表达后人胆管癌细胞株HUCCA-1细胞功能、PI3KCA及Akt1的蛋白及mRNA表达水平改变,探讨Gab1、PI3KCA及Akt1表达在胆管癌恶性行为中的作用机制。方法 Gab1siRNA转染HUCCA-1细胞,采用qRT-PCR法及Western blot法检测转染效率及PI3KCA、Akt1表达水平,MTT检测转染后细胞增殖变化,流式细胞术检测转染后细胞凋亡变化,Transwell检测细胞迁移及侵袭能力。结果 Gab1siRNA在HUCCA-1细胞中的转染效率约为65%~70%,转染效率佳;Gab1siRNA转染HUCCA-1细胞后,PI3KCA、Akt1的蛋白及mRNA表达量下调;Gab1siRNA组HUCCA-1细胞增殖能力在48 h、72 h及96 h均低于siRNA control组和Mock组(P〈0.05);Gab1siRNA组的HUCCA-1细胞凋亡率高于Mock及siRNA control组(P〈0.05);Gab1siRNA组HUCCA-1细胞迁移减少百分比及侵袭能力低于siRNA control组及Mock组(P<0.05)。结论 Gab1可能通过激活PI3K/Akt信号通路表达促进肿瘤细胞恶性行为,Gab1可作为胆管癌治疗新的靶向标志物及候选基因。 展开更多
关键词 胆管癌 增殖 凋亡 Gab1 hucca-1 PI3K/AKT信号通路
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Focal adhesion kinase and Src phosphorylations in HGF-induced proliferation and invasion of human cholangiocarcinoma cell line, HuCCA-1 被引量:5
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作者 Urai Pongchairerk Jun-Lin Guan Vijittra Leardkamolkarn 《World Journal of Gastroenterology》 SCIE CAS CSCD 2005年第37期5845-5852,共8页
AIM: To study the role of focal adhesion kinase (FAK) and its association with Src in hepatocyte growth factor (HGF)-induced cell signaling in cholangiocarcinoma progression.METHODS: Previously isolated HuCCA-1 cells ... AIM: To study the role of focal adhesion kinase (FAK) and its association with Src in hepatocyte growth factor (HGF)-induced cell signaling in cholangiocarcinoma progression.METHODS: Previously isolated HuCCA-1 cells were re-characterized by immunofluorescent staining and reverse transcriptase-polymerase chain reaction assay for the expression of cytokeratin 19, HGF and c-Met mRNA. Cultured HuCCA-1 cells were treated with HGF and determined for cell proliferation and invasion effects by MTT and invasion assays. Western blotting, immunoprecipitation, and co-immunoprecipitation were also performed to study the phosphorylation and interaction of FAK and Src. A novel Src inhibitor (AZM555130) was applied in cultures to investigate the effects on FAK phosphorylation inhibition and on cell proliferation and invasion.RESULTS: HGF enhanced HuCCA-1 cell proliferation and invasion by mediating FAK and Src phosphorylations.FAK-Src interaction occurred in a time-dependent manner that Src was proved to be an upstream signaling molecule to FAK. The inhibitor to Src decreased FAK phosphorylation level in correlation with the reduction of cell proliferation and invasion.CONCLUSION: FAK plays a significant role in signaling pathway of HGF-responsive cell line derived from cholangiocarcinoma. Autophosphorylated Src, induced by HGF, mediates Src kinase activation, which subsequently phosphorylates its substrate, FAK, and signals to cell proliferation and invasion. 展开更多
关键词 Human cholangiocarcinoma Hepatocyte growth factor C-MET Focal adhesion kinase SRC
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