ABSTRACT

The roles of lncRNA TSLNC8 and its synergetic effects with osimertinib remain unknown in lung cancer. qRT-PCR or western blotting was performed to determine the expression levels of TSLNC8, EGFR and STAT3. Colony formation and MTT assays were used to evaluate cell proliferation. Transwell and wound healing assays were performed to assess migration and invasion abilities. Flow cytometry with Annexin V/PI staining was used to detect changes in cell apoptosis. Nude mice subcutaneous tumor model was constructed and used for validating the effects of TSLNC8 and osimertinib in vivo. Expression of TSLNC8 was down-regulated in clinical lung cancer tissues and cell lines. TSLNC8 overexpression or osimertinib administration led to promotion of apoptosis and inhibition of cell proliferation, migration and invasion, as well as deactivation of the EGFR-STAT3 pathway, whereas TSLNC8 knockdown had opposite effects. Moreover, the above effects of osimertinib were remarkably enhanced by TSLNC8 overexpression and inhibited by TSLNC8 knockdown, respectively. Meanwhile, the effects of TSLNC8 overexpression were reversed by STAT3 activation or EGFR overexpression. In the animal model, combination of TSLNC8 overexpression and osimertinib administration resulted in efficient suppression of tumor growth. In this study, we revealed a TSLNC8-EGFR-STAT3 signaling axis in lung cancer, and TSLNC8 overexpression significantly enhanced the anti-tumor effects of osimertinib via inhibiting EGFR-STAT3 signaling.

摘要

lncRNA TSLNC8 的作用及其与奥希替尼的协同作用在肺癌中仍然未知。进行 qRT-PCR 或蛋白质印迹以确定 TSLNC8、EGFR 和 STAT3 的表达水平。集落形成和 MTT 测定用于评估细胞增殖。进行 Transwell 和伤口愈合试验以评估迁移和侵袭能力。使用 Annexin V/PI 染色的流式细胞术检测细胞凋亡的变化。构建裸鼠皮下肿瘤模型并用于验证TSLNC8和奥希替尼在体内的作用. TSLNC8 的表达在临床肺癌组织和细胞系中被下调。TSLNC8 过表达或奥希替尼给药导致促进细胞凋亡和抑制细胞增殖、迁移和侵袭，以及 EGFR-STAT3 通路的失活，而 TSLNC8 敲低具有相反的效果。此外，奥希替尼的上述作用分别被 TSLNC8 过表达显着增强和被 TSLNC8 敲低抑制。同时，STAT3 激活或 EGFR 过表达逆转了 TSLNC8 过表达的影响。在动物模型中，TSLNC8 过表达和奥希替尼给药的组合导致肿瘤生长的有效抑制。在这项研究中，我们揭示了肺癌中的 TSLNC8-EGFR-STAT3 信号轴，