School of Medicine, Yunnan University, Kunming, Yunnan, China.
Eur Rev Med Pharmacol Sci. 2020 Sep;24(18):9497-9510. doi: 10.26355/eurrev_202009_23035.
Orthodenticle Homeobox 1 (OTX1) has been found to be closely related to the development of several human tumours. However, the function and underlying molecular mechanisms of OTX1 in non-small cell lung cancer (NSCLC) are unclear. This research was performed to investigate the effects of downregulating OTX1 gene expression on the proliferation, migration, invasion, cell cycle and apoptosis of human NSCLC cell lines.
Cultured NCI-H292 and XWLC cells were transfected with control small interfering RNA (siNC) or experimental siRNA (siOTX1). The mRNA levels were detected using a quantitative real-time PCR (RT-qPCR) assay. A Cell Counting Kit-8 (CCK-8) and a Real Time Cell Analyzer (RTCA) were used to determine cell activity. The RTCA and transwell chambers were used to assess cell migration and invasion. In addition, cell cycle progression and apoptosis were measured using flow cytometry, and the expression levels of key signalling pathway proteins were examined by Western blotting.
The results revealed that compared with the control group, the experimental group exhibited significantly decreased cell activity (***p<0.001), significantly decreased migration and invasion abilities (***p<0.001), and cell cycle arrest in G2/M phase (*p<0.05). However, the number of apoptotic cells was higher in the experimental group than in the control group (*p<0.05). The Western blotting results were consistent with the functional experiment results.
Silencing the OTX1 gene suppressed the proliferation, migration and invasion of NCI-H292 and XWLC cells, impeded the cell cycle transition from G2 to M phase, and accelerated apoptosis, revealing OTX1, a regulator of NSCLC, as a potential new therapeutic target.
Orthodenticle Homeobox 1(OTX1)已被发现与多种人类肿瘤的发生密切相关。然而,OTX1 在非小细胞肺癌(NSCLC)中的功能和潜在分子机制尚不清楚。本研究旨在探讨下调 OTX1 基因表达对人 NSCLC 细胞系增殖、迁移、侵袭、细胞周期和凋亡的影响。
培养 NCI-H292 和 XWLC 细胞,用对照小干扰 RNA(siNC)或实验性 siRNA(siOTX1)转染。采用实时定量 PCR(RT-qPCR)检测 mRNA 水平。用细胞计数试剂盒-8(CCK-8)和实时细胞分析(RTCA)测定细胞活性。用 RTCA 和 Transwell 室评估细胞迁移和侵袭。此外,用流式细胞术检测细胞周期进程和凋亡,并用 Western blot 检测关键信号通路蛋白的表达水平。
与对照组相比,实验组细胞活性显著降低(***p<0.001),迁移和侵袭能力显著降低(***p<0.001),G2/M 期细胞周期阻滞(*p<0.05)。然而,实验组细胞凋亡数量高于对照组(*p<0.05)。Western blot 结果与功能实验结果一致。
沉默 OTX1 基因抑制了 NCI-H292 和 XWLC 细胞的增殖、迁移和侵袭,阻碍了细胞周期从 G2 向 M 期的过渡,并加速了凋亡,提示 OTX1 作为 NSCLC 的调节因子,可能成为一个新的潜在治疗靶点。