Department of Breast Surgery, Affliliated Sanming First Hospital of Fujian Medical University, Sanming City, Fujian Province, China.
Department of Thyroid and Breast Surgery, Nanping First Hospital Affiliated to Fujian Medical University, Nanping City, Fujian Province, China.
J Obstet Gynaecol Res. 2023 Aug;49(8):2175-2184. doi: 10.1111/jog.15710. Epub 2023 Jun 20.
KIFC1 exerts an important function in centrosome aggregation in breast cancer (BC) cells and a variety of other cancer cells, but its potential mechanisms in BC pathogenesis are yet fully elucidated. The aim of this study was to investigate the effects of KIFC1 on BC progression and its underlying mechanisms.
Expression of ELK1 and KIFC1 in BC was analyzed by The Cancer Genome Atlas database and quantitative real-time polymerase chain reaction. Cell proliferative capacity was examined by CCK-8 and colony formation assays, respectively. Glutathione (GSH)/glutathione disulfide (GSSG) ratio and GSH level were measured using the kit. Expression of GSH metabolism-related enzymes (G6PD, GCLM, and GCLC) was detected by western blot. Intracellular reactive oxygen species (ROS) levels were measured by the ROS Assay Kit. The transcription factor ELK1 upstream of KIFC1 was identified by hTFtarget, KnockTFv2 database and Pearson correlation. Their interaction was validated by dual-luciferase reporter assay and chromatin immunoprecipitation.
This study demonstrated the upregulation of ELK1 and KIFC1 in BC and found that ELK1 could bind to the KIFC1 promoter to promote KIFC1 transcription. KIFC1 overexpression increased cell proliferation and intracellular GSH levels, while decreasing intracellular ROS levels. The addition of the GSH metabolism inhibitor BSO attenuated the promotion of BC cell proliferation induced by KIFC1 overexpression. In addition, KIFC1 overexpression reversed the inhibitory effect of knockdown of ELK1 on BC cell proliferation.
ELK1 was a transcriptional factor of KIFC1. ELK1/KIFC1 axis reduced ROS level by increasing GSH synthesis, thus facilitating BC cell proliferation. Current observations suggest that ELK1/ KIFC1 may be a potential therapeutic target for BC treatment.
KIFC1 在乳腺癌（BC）细胞和多种其他癌细胞的中心体聚集中发挥重要功能，但它在 BC 发病机制中的潜在机制尚未完全阐明。本研究旨在探讨 KIFC1 对 BC 进展的影响及其潜在机制。
通过癌症基因组图谱数据库和实时定量聚合酶链反应分析 BC 中 ELK1 和 KIFC1 的表达。分别通过 CCK-8 和集落形成实验检测细胞增殖能力。使用试剂盒测量谷胱甘肽（GSH）/谷胱甘肽二硫化物（GSSG）比和 GSH 水平。通过 Western blot 检测 GSH 代谢相关酶（G6PD、GCLM 和 GCLC）的表达。通过 ROS 测定试剂盒测量细胞内活性氧（ROS）水平。通过 hTFtarget、KnockTFv2 数据库和 Pearson 相关性鉴定 KIFC1 上游的转录因子 ELK1。通过双荧光素酶报告基因检测和染色质免疫沉淀验证它们的相互作用。
本研究表明 ELK1 和 KIFC1 在 BC 中上调，并发现 ELK1 可以结合到 KIFC1 启动子上，促进 KIFC1 转录。KIFC1 过表达增加细胞增殖和细胞内 GSH 水平，同时降低细胞内 ROS 水平。添加 GSH 代谢抑制剂 BSO 减弱了 KIFC1 过表达诱导的 BC 细胞增殖的促进作用。此外，KIFC1 过表达逆转了 ELK1 敲低对 BC 细胞增殖的抑制作用。
ELK1 是 KIFC1 的转录因子。ELK1/KIFC1 轴通过增加 GSH 合成降低 ROS 水平，从而促进 BC 细胞增殖。目前的观察结果表明，ELK1/KIFC1 可能是 BC 治疗的潜在治疗靶点。
