离子通道蛋白Piezo1在人源非小细胞肺癌中的功能研究  

Functional Study of Ion Channel Protein Piezo1 in Human Non-small Cell Lung Cancer

作　　者：赵敏萱 夏凡晨 刘晨 张文悦 石金磊 康宇佳 Zhao Minxuan;Xia Fanchen;Liu Chen;Zhang Wenyue;Shi Jinlei;Kang Yujia(chool of Life Science and Technology,Shanghai Tech University,Shanghai 201210,China)

机构地区：[1]上海科技大学生命科学与技术学院,上海201210

出　　处：《黑龙江科学》2023年第12期86-93,共8页Heilongjiang Science

摘　　要：为探讨Piezo1基因敲减对非小细胞肺癌A549细胞增殖、迁移及侵袭的影响,通过siRNA转染构建Piezo1敲减A549细胞系,经RT-qPCR、Western blot检测其沉默效果,Piezo1的表达在mRNA水平与蛋白水平都显著下调。通过CCK8法检测敲减后细胞增殖能力变化,通过划痕修复实验检测细胞迁移能力,通过Transwell小室实验检测细胞侵袭能力,通过Fluo-4 AM Ca^(2+)探针成像实验检测胞内Ca^(2+)含量,通过RT-qPCR与Western blot法检测Piezo1下游信号通路相关基因的转录水平及蛋白磷酸化水平。结果表明,Piezo1靶向siRNA能够下调A549细胞系中Piezo1 mRNA及蛋白表达,抑制A549细胞的增殖、迁移、侵袭能力,降低胞内Ca^(2+)水平,导致增殖相关基因Myc、Cyclin-D1、Cyclin-E、CDK2表达水平下调,迁移相关基因E-cadherin、N-cadherin、Vimentin表达水平上调,Ca^(2+)相关信号通路蛋白Akt的磷酸化水平下调,4EBP1蛋白的磷酸化水平显著提高,说明Piezo1基因敲减抑制A549细胞的增殖、迁移及侵袭能力,为非小细胞肺癌的治疗提供了新的分子靶标。To investigate the effects of Piezo 1 gene knockout on the proliferation,migration and invasion of non-small cell lung cancer A549 cells,a Piezo 1 knockout A549 cell line is constructed by siRNA transfection,and the silencing effect is detected by RT-qPCR and Western blot.The expression of Piezo 1 is significantly down-regulated at both mRNA and protein levels.Changes in cell proliferation capacity after knockdown are detected by CCK8 assay,cell migration capacity is detected by scratch repair assay,cell invasion capacity is detected by Transwell chamber assay,intracellular Ca 2+content is detected by Fluo-4 AM Ca 2+probe imaging assay,and transcriptional levels and protein phosphorylation levels of genes related to the downstream signaling pathway of Piezo 1 are detected by RT-qPCR and Western blot.The results show that Piezol-targeted siRNA can down-regulate the mRNA and protein expression of Piezo1 in A549 cell line,inhibit the proliferation,migration and invasion ability of A549 cells,and reduce the intracellular Ca 2+level,resulting in down-regulated expression levels of proliferation-related genes Myc,Cyclin-D 1,Cyclin-E and CDK 2.The expression levels of migration-related genes E-cadherin,N-cadherin and Vimentin are up-regulated,the phosphorylation level of Ca 2+related signaling pathway protein Akt is down-regulated,and the phosphorylation level of 4EBP1 protein is significantly increased,suggesting that the knockdown of Piezo1 gene inhibiting the proliferation,migration and invasion of A549 cells.This provides a new molecular target for the treatment of non-small cell lung cancer.

关 键 词：非小细胞肺癌 Piezo1 A549细胞系 增殖 迁移 侵袭 

分 类 号：R734.2[医药卫生—肿瘤]