Han Qian, Cheng Peng, Yang Hongjie, Liang Hengpo, Lin Fengchun
Department of Radiotherapy, Henan Provincial People's Hospital, People's Hospital of Zhengzhou University, Zhengzhou, Henan, China.
J Cell Biochem. 2020 Aug;121(8-9):3901-3912. doi: 10.1002/jcb.29554. Epub 2019 Nov 11.
Epithelial-mesenchymal transformation (EMT) is associated with drug resistance in human lung adenocarcinoma cells, but its specific mechanism has not been clarified. In this study, we investigated the effect of miRNA-146b on EMT in cisplatin (DDP) resistant human lung adenocarcinoma cells and the corresponding mechanism. Cisplatin resistant (CR) human lung adenocarcinoma cells (A549/DDP and H1299/DDP) were established, and the EMT characteristics and invasion and metastasis ability of CR cells were determined by tumor cell-related biological behavior experiments. The role of miR-146b in EMT of CR cells was determined by in vitro functional test. The targeted binding of miR-146b to protein tyrosine phosphatase 1B (PTP1B) was verified by biological information and double luciferin gene reporting experiments. The effect of miR-146b on tumor growth and EMT phenotype in vivo was investigated by establishing the xenotransplantation mouse model. Compared with the control group, H1299/DDP and A549/DDP cells showed the enhanced EMT phenotypes, invasion and migration ability. Besides, miR-146b was lowly expressed in H1299/DDP and A549/DDP cells. More importantly, overexpressed miR-146b could specifically bind to PTP1B, thus inhibiting the EMT process and ultimately reducing CR in H1299/DDP and A549/DDP cells. Finally, overexpressed miR-146b observably inhibited tumor growth in xenograft model mice and inhibited the EMT phenotype of A549/DDP cells in vivo by regulating the expressions of EMT-related proteins. Overexpressed miR-146b could reverse the EMT phenotype of CR lung adenocarcinoma cells by targeting PTP1B, providing new therapeutic directions for CR of lung adenocarcinoma cells.
上皮-间质转化（EMT）与人类肺腺癌细胞的耐药性相关，但其具体机制尚未阐明。在本研究中，我们探究了miRNA-146b对顺铂（DDP）耐药的人类肺腺癌细胞中EMT的影响及其相应机制。建立了顺铂耐药（CR）的人类肺腺癌细胞系（A549/DDP和H1299/DDP），并通过肿瘤细胞相关生物学行为实验确定CR细胞的EMT特征以及侵袭和转移能力。通过体外功能试验确定miR-146b在CR细胞EMT中的作用。通过生物信息学和双荧光素酶基因报告实验验证miR-146b与蛋白酪氨酸磷酸酶1B（PTP1B）的靶向结合。通过建立异种移植小鼠模型研究miR-146b对体内肿瘤生长和EMT表型的影响。与对照组相比，H1299/DDP和A549/DDP细胞表现出增强的EMT表型、侵袭和迁移能力。此外，miR-146b在H1299/DDP和A549/DDP细胞中低表达。更重要的是，过表达的miR-146b可特异性结合PTP1B，从而抑制EMT过程，并最终降低H1299/DDP和A549/DDP细胞中的CR。最后，过表达的miR-146b在异种移植模型小鼠中显著抑制肿瘤生长，并通过调节EMT相关蛋白的表达在体内抑制A549/DDP细胞的EMT表型。过表达的miR-146b可通过靶向PTP1B逆转CR肺腺癌细胞的EMT表型，为肺腺癌细胞的CR提供新的治疗方向。
