Background: Colorectal cancer (CRC) is a prevalent form of cancer globally, characterized by a high mortality rate. Therefore, discovering effective therapeutic approaches for CRC treatment is critical. Methods: The levels of KIF20A in CRC clinical samples were determined using Western Blot and immunofluorescence assay. SW480 cells were transfected with siRNA targeting KIF20A, while HT-29 cells were transfected with a KIF20A overexpression vector. Cell viability and apoptosis of CRC cells were assessed using CCK-8 and TUNEL analysis. Migration ability was investigated using Transwell. The levels of pyruvate, lactate and ATP were determined through corresponding assay kits. Western Blot was applied to confirm the level of proteins associated with glycolysis, c- Myc, HIF-1α, PKM2 and LDHA. Subsequently, functional rescue experiments were conducted to investigate further the regulatory relationship between KIF20A, c-Myc, and HIF-1α in colorectal cancer (CRC), employing the c-Myc inhibitor 10058-F4 and c-Myc overexpression plasmids. Results: KIF20A was up-regulated in vivo and in vitro in CRC. KIF20A knockdown inhibited cell viability and migration while promoting cell apoptosis in SW480 cells. Conversely, overexpression of KIF20A yielded contrasting effects in HT-29 cells. Moreover, inhibition of KIF20A restrained the pyruvate, lactate production and ATP level, whereas overexpression of KIF20A enhanced the Warburg effect. Western Blot indicated that knockdown KIF20A attenuated the levels of c-Myc, HIF-1α, PKM2 and LDHA. In addition, rescue experiments further verified that KIF20A enhanced the Warburg effect by the KIF20A/c-Myc/HIF-1α axis in CRC. Conclusion: KIF20A, being a crucial regulator in the progression of CRC, has the potential to be a promising therapeutic target for the treatment of CRC.

中文翻译：

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KIF20A 通过 C-Myc/HIF-1α 轴促进 CRC 进展和 Warburg 效应

背景：结直肠癌（CRC）是全球流行的癌症形式，其特点是死亡率高。因此，寻找有效的CRC治疗方法至关重要。方法：采用Western Blot和免疫荧光法测定CRC临床样本中KIF20A的水平。SW480细胞用靶向KIF20A的siRNA转染，而HT-29细胞用KIF20A过表达载体转染。使用 CCK-8 和 TUNEL 分析评估 CRC 细胞的细胞活力和凋亡。使用 Transwell 研究迁移能力。通过相应的检测试剂盒测定丙酮酸、乳酸和ATP的水平。应用蛋白质印迹来确认与糖酵解、c-Myc、HIF-1α、PKM2 和 LDHA 相关的蛋白质水平。随后，利用 c-Myc 抑制剂 10058-F4 和 c-Myc 过表达质粒进行功能拯救实验，进一步研究结直肠癌 (CRC) 中 KIF20A、c-Myc 和 HIF-1α 之间的调控关系。结果：KIF20A 在 CRC 体内和体外均上调。KIF20A 敲低抑制 SW480 细胞中的细胞活力和迁移，同时促进细胞凋亡。相反，KIF20A 的过度表达在 HT-29 细胞中产生了相反的效果。此外，KIF20A 的抑制抑制了丙酮酸、乳酸的产生和 ATP 水平，而 KIF20A 的过度表达增强了 Warburg 效应。Western Blot 表明敲低 KIF20A 会减弱 c-Myc、HIF-1α、PKM2 和 LDHA 的水平。此外，挽救实验进一步证实KIF20A通过KIF20A/c-Myc/HIF-1α轴增强CRC中的Warburg效应。结论：KIF20A是CRC进展的关键调节因子，有可能成为CRC治疗的有前景的治疗靶点。