吡非尼酮与仑伐替尼联合应用对人胆管癌细胞系HuCCT1的增殖克隆抑制作用及其机制  

Inhibitory effects of pirfenidone combined with lenvatinib on proliferation and colony formation of human cholangiocarcinoma HuCCT1 cells

作　　者：李丽萍 蒲诗云 任常谕 周永杰[2] 周后凤[1] LI Liping;PU Shiyun;REN Changyu;ZHOU Yongjie;ZHOU Houfeng(Department of Pharmacy,Chengdu Fifth People's Hospital,Chengdu 611130,China)

机构地区：[1]成都市第五人民医院药剂科,成都611130 [2]四川大学华西医院卫生部移植工程与移植免疫重点实验室病理研究室

出　　处：《山东医药》2022年第24期1-5,共5页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(82003828)。

摘　　要：目的观察吡非尼酮(PFD)联合仑伐替尼(Len)对人胆管癌HuCCT1细胞增殖、克隆形成、自噬的影响,并探讨其可能机制。方法人胆管癌HuCCT1细胞经PFD、Len或PFD联合Len处理后,CCK-8法检测各组细胞增殖能力,平板克隆形成实验检测细胞克隆形成能力,免疫荧光染色检测细胞自噬体数目,蛋白免疫印迹检测AMPK、mTOR、S6蛋白磷酸化水平及自噬标记蛋白LC3-Ⅱ蛋白水平。结果与PFD单药组和Len单药组相比,联合组的细胞增殖活力、克隆形成数目、LC3-Ⅱ表达水平及自噬体形成数目都显著降低,P均<0.05;与PFD单药组和Len单药组相比,联合组的AMPK蛋白磷酸化水平升高,mTOR、S6蛋白磷酸化水平降低,P均<0.05。结论PFD联合Len可抑制HuCCT1细胞增殖、克隆形成,机制可能是其通过上调AMPK磷酸化水平,下调mTOR的磷酸化水平,进而抑制mTOR信号通路,促进细胞自噬。Objective To investigate the inhibitory effects of pirfenidone(PFD)combined with lenvatinib(Len)on proliferation and colony formation of human cholangiocarcinoma HuCCT1 cells and their molecular mechanisms.Methods Human cholangiocarcinoma HuCCT1 cells were treated with PFD,Len,or their combination.After treatment,the proliferation and colony formation of cells were analyzed by CCK-8 and clonal assay,respectively;the number of autophagosomes was detected by immunofluorescence;the protein phosphorylation levels of AMPK,mTOR,and S6,and the level of LC3-Ⅱprotein expression were assessed by Western blotting.Results Compared with the PFD group and Len group,the combination group of PFD and Len significantly inhibited the proliferation and colony formation of HuCCT1 cells,upregulated the protein level of LC3-Ⅱand the autophagosome formation of HuCCT1 cells(all P<0.05).Additionally,compared with the PFD group and Len group,the combination group of PFD and Len up-regulated the phosphorylation level of AMPK protein,and down-regulated the phosphorylation levels of mTOR and S6 protein(all P<0.05).Conclusions PFD combined with Len can inhibit the proliferation and colony formation of HuCCT1 cells.The mechanism may be that they increase the phosphorylation level of AMPK and down-regulate the phosphorylation level of mTOR,thereby inhibiting the mTOR signaling pathway and promoting autophagy.

关 键 词：吡非尼酮 仑伐替尼 胆管癌 HuCCT1细胞 细胞增殖 细胞克隆形成 细胞自噬 

分 类 号：R735.8[医药卫生—肿瘤]