Wang Yan, Wang XiaoLi, Xu Jun, Zhang Didi, Cao YongFeng
Department of Medical Oncology, Tumor Hospital Affiliated to Nantong University & Nantong Tumor Hospital, Nantong, Jiangsu, China.
Pharmacogenomics. 2025 Apr-Apr;26(5-6):165-170. doi: 10.1080/14622416.2025.2504864. Epub 2025 May 14.
To explore the impact of low miR-369-3p expression on the resistance of PC-9 cells to osimertinib.
The PC-9/AZD9291 cell line was established with osimertinib. Real-time quantitative PCR was employed to measure the expression levels of miR-369-3p in both PC-9 and PC-9/AZD9291 cells, and Western blotting was utilized to detect PTPN12 protein expression. A dual-luciferase reporter assay was conducted to investigate the target relationship between miR-369-3p and PTPN12. CCK8 assays were performed to evaluate the impact of miR-369-3p inhibition on drug resistance.
In comparison to PC-9 cells, there was a significant upregulation of miR-369-3p and downregulation of PTPN12 protein in PC-9/AZD9291 cells ( < 0.05). Transfection with the miR-369-3p inhibitor resulted in decreased levels of miR-369-3p and increased expression of PTPN12 protein in PC-9/AZD9291 cells ( < 0.05). Conversely, transfection with miR-369 mimics led to an increase in miR-369-3p levels accompanied by a decrease in PTPN12 protein ( < 0.05). Notably, treatment with the miR-369-3p inhibitor lowered the IC50 value for PC-9/AZD9291 cells; however, following downregulation of PTPN12 using PTPN12-siRNA, sensitivity due to low expression of miR-369-3p was significantly diminished ( < 0.05).
miR-369-3p plays a crucial role in modulating drug resistance in PC-9/AZD9291 cells against osimertinib through regulation of PTPN12.
探讨低表达的miR-369-3p对PC-9细胞对奥希替尼耐药性的影响。
用奥希替尼建立PC-9/AZD9291细胞系。采用实时定量PCR检测PC-9和PC-9/AZD9291细胞中miR-369-3p的表达水平，并用蛋白质免疫印迹法检测PTPN12蛋白表达。进行双荧光素酶报告基因检测以研究miR-369-3p与PTPN12之间的靶向关系。采用CCK8法评估抑制miR-369-3p对耐药性的影响。
与PC-9细胞相比，PC-9/AZD9291细胞中miR-369-3p显著上调，PTPN12蛋白下调（<0.05）。用miR-369-3p抑制剂转染导致PC-9/AZD9291细胞中miR-369-3p水平降低，PTPN12蛋白表达增加（<0.05）。相反，用miR-369模拟物转染导致miR-369-3p水平升高，同时PTPN12蛋白减少（<0.05）。值得注意的是，用miR-369-3p抑制剂处理降低了PC-9/AZD9291细胞的IC50值；然而，使用PTPN12-siRNA下调PTPN12后，由于miR-369-3p低表达导致的敏感性显著降低（<0.05）。
miR-369-3p通过调节PTPN12在调节PC-9/AZD9291细胞对奥希替尼的耐药性中起关键作用。
