Lei Quan, Liu Ping, Guan Xinlei, Liu Li, He Wenjuan
Department of Pharmacy, Wuhan Fourth Hospital, No. 473 Hanzheng Street, Qiaokou District, Wuhan, 430030 China.
Cytotechnology. 2025 Feb;77(1):23. doi: 10.1007/s10616-024-00673-8. Epub 2024 Dec 19.
Osimertinib has been demonstrated to be effective for improving the prognosis of patients with epidermal growth factor receptor mutation-positive lung cancer. However, osimertinib resistance inevitably emerges throughout the treatment course. This study explored the function and mechanism of long noncoding RNA LINC01278 in osimertinib-resistant NSCLC cells. Osimertinib-resistant non-small cell lung cancer (NSCLC) cells were established by treating PC9 and HCC827 cells with increasing doses of osimertinib for over 6 months. LINC01278 expression in parental and drug-resistant cells (PC9-OR and HCC827-OR) was measured by polymerase chain reaction. Cell counting kit 8 assays were used to examine cell viability and half-maximal inhibitory concentration values. The effects of LINC01278 knockdown on cell proliferation and apoptosis were measured by colony formation assays and flow cytometry. A luciferase reporter assay was performed to verify the interaction between LINC01278 and miR-324-3p or the binding ability between miR-324-3p and ZFX. Protein levels of ZFX and apoptotic markers in NSCLC cells were measured by western blotting. As shown by experimental results, LINC01278 was highly expressed in osimertinib-resistant NSCLC cells compared to its expression in parental cells. The silencing of LINC01278 improved the sensitivity of drug-resistant cells towards osimertinib. LINC1278 depletion inhibited osimertinib-resistant cell proliferation while promoting cell apoptosis. LINC01278 interacted with miR-324-3p to regulate ZFX expression. ZFX could be targeted by miR-324-3p in PC9-OR and HCC827-OR cells. ZFX overexpression counteracted the suppressive impact of LINC01278 silencing on the malignant behavior of PC9-OR and HCC827-OR cells. In conclusion, LINC01278 knockdown alleviates osimertinib resistance of NSCLC cells by regulating downstream miR-324-3p and ZFX.
The online version contains supplementary material available at 10.1007/s10616-024-00673-8.
奥希替尼已被证明可有效改善表皮生长因子受体突变阳性肺癌患者的预后。然而，在整个治疗过程中不可避免地会出现奥希替尼耐药。本研究探讨了长链非编码RNA LINC01278在奥希替尼耐药非小细胞肺癌细胞中的功能及机制。通过用递增剂量的奥希替尼处理PC9和HCC827细胞超过6个月，建立了奥希替尼耐药的非小细胞肺癌（NSCLC）细胞。通过聚合酶链反应检测亲本细胞和耐药细胞（PC9-OR和HCC827-OR）中LINC01278的表达。使用细胞计数试剂盒8检测来检测细胞活力和半数最大抑制浓度值。通过集落形成试验和流式细胞术检测LINC01278敲低对细胞增殖和凋亡的影响。进行荧光素酶报告基因试验以验证LINC01278与miR-324-3p之间的相互作用或miR-324-3p与ZFX之间的结合能力。通过蛋白质印迹法检测NSCLC细胞中ZFX和凋亡标志物的蛋白水平。实验结果表明，与亲本细胞相比，LINC01278在奥希替尼耐药的NSCLC细胞中高表达。LINC01278的沉默提高了耐药细胞对奥希替尼的敏感性。LINC1278的缺失抑制了奥希替尼耐药细胞的增殖，同时促进了细胞凋亡。LINC01278与miR-324-3p相互作用以调节ZFX的表达。在PC9-OR和HCC827-OR细胞中，ZFX可被miR-324-3p靶向。ZFX的过表达抵消了LINC01278沉默对PC9-OR和HCC827-OR细胞恶性行为的抑制作用。总之，LINC01278的敲低通过调节下游的miR-324-3p和ZFX减轻了NSCLC细胞的奥希替尼耐药性。
在线版本包含可在10.1007/s10616-024-00673-8获取的补充材料。
