Zhonghua zhong liu za zhi [Chinese journal of oncology] ·23 Nov 2026 ·Added 11 Nov 2026

[Kinesin family member 26A inhibits colorectal cancer cell proliferation, invasion, and migration by regulating the JunB-mediated PI3K-AKT signaling pathway].

M J Li, L L Zheng, N X Ren, R Q Yan, S S Zhang, D X Lin, W Tan

Biochemistry

DOI ↗

Abstract

Objective: To investigate the regulatory role of Kinesin family member 26A (KIF26A) in colorectal cancer (CRC) cell invasion and migration and elucidate its underlying molecular mechanism. Methods: Bioinformatics analysis using TCGA databases was performed to assess the correlation between KIF26A expression and clinic pathological characteristics. Flow cytometry was employed to examine the effect of KIF26A on cell cycle distribution. Transwell assays were conducted to evaluate the impact of KIF26A on CRC cell invasion and migration. RNA-seq, gene set enrichment analysis (GSEA), quantitative polymerase chain reaction (qPCR), Western blot, Co-Immunoprecipitation (Co-IP)and Chromatin Immunoprecipitation followed by qPCR (ChIP-qPCR) were utilized to explore the downstream regulatory network and molecular mechanisms. Results: Following KIF26A knockdown, the G0/G1 phase proportion in HCT116 cells decreased from (75.80±0.64)% in the negative control group to (73.57±0.53)% and (72.17±0.08)% in the shKIF26A#1 and shKIF26A#2 groups, respectively (both P＜0.05); the number of invasive cells increased from 192.60±4.50 to 483.00±6.74 and 471.80±8.01, respectively (both P＜0.001); and the number of migratory cells increased from 288.40±4.39 to 489.20±5.58 and 490.40±7.17, respectively (both P＜0.001). In RKO cells, the G0/G1 phase proportion decreased from (61.17±0.41)% in the negative control group to (57.13±0.23)% and (57.70±0.06)% in the shKIF26A#1 and shKIF26A#2 groups, respectively (both P＜0.001); the number of invasive cells increased from 57.60±4.11 to 160.80±5.48 and 136.20±4.44, respectively (both P＜0.001); and the number of migratory cells increased from 81.00±2.95 to 270.80±4.42 and 299.00±3.63, respectively (both P＜0.001). Following KIF26A overexpression, the G0/G1 phase proportion in HCT116 cells increased from (41.20±0.88)% in the empty vector control group to (54.33±0.33)% in the KIF26A overexpression group (P<0.001); the number of invasive cells decreased from 338.00±5.57 to 194.80±4.90 (P＜0.001); and the number of migratory cells decreased from 408.60±3.33 to 263.60±3.59 (P＜0.001). In RKO cells, the G0/G1 phase proportion increased from (57.63±0.33)% in the empty vector control group to (62.65±0.45)% in the KIF26A overexpression group (P＜0.001); the number of invasive cells decreased from 102.60±3.37 to 54.80±2.01 (P＜0.001); and the number of migratory cells decreased from 180.00±4.18 to 90.40±4.77 (P＜0.001). GSEA of differentially expressed genes between KIF26A-high and KIF26A-low expression groups in TCGA samples, as well as between KIF26A-overexpressing RKO cells and empty vector controls, revealed significant enrichment of differentially expressed genes in pathways related to cell cycle, invasion, and migration (all P＜0.05). Mechanistic studies revealed that KIF26A knockdown suppressed the expression of retinoblastoma protein (RB) and E2F transcription factor 1 (E2F1) in colon cancer cells while promoting the expression of phosphatidylinositol 3-kinase (PI3K) and AKT serine/threonine kinase 1 (AKT1); conversely, KIF26A overexpression promoted RB and E2F1 expression and inhibited PI3K and AKT1 expression. Co-IP combined with mass spectrometry identified JunB as an interacting protein of KIF26A. ChIP assay results showed that JunB binds to the promoter region of PTEN. Rescue experiments demonstrated that JunB knockdown partially reversed the inhibition of invasion and migration mediated by KIF26A overexpression (P＜0.001). Conclusions: KIF26A regulates cell cycle progression through the RB/E2F1 pathway and modulates the PI3K/AKT signaling pathway via interaction with JunB, thereby inhibiting the invasion and migration of colorectal cancer cells. The elucidation of the KIF26A-JunB-PI3K-AKT regulatory axis provides a novel perspective for understanding the mechanisms of tumor metastasis, and KIF26A may serve as a potential predictive biomarker and therapeutic target for tumor metastasis. 目的：探讨驱动蛋白家族成员26A（KIF26A）对结直肠癌细胞侵袭、迁移的调控作用及其分子机制。方法：利用癌症基因组图谱（TCGA）数据库中498例结直肠癌患者的资料，分析KIF26A RNA表达与临床病理特征的关系。在人结直肠癌细胞系HCT116和RKO中敲低和过表达KIF26A，流式细胞术检测细胞周期分布，Transwell实验评估细胞侵袭和迁移能力，通过转录组测序、基因集富集分析（GSEA）、反转录定量聚合酶链反应、蛋白质免疫印迹法、免疫共沉淀（Co-IP）及染色质免疫沉淀（ChIP）等实验技术探究KIF26A的下游分子调控网络及分子机制。结果：敲低KIF26A后，HCT116细胞G0/G1期比例由阴性对照组的（75.80±0.64）%下降至shKIF26A#1组和shKIF26A#2组的（73.57±0.53）%和（72.17±0.08）%（均P＜0.05），侵袭细胞数由（192.60±4.50）个上升至（483.00±6.74）个和（471.80±8.01）个（均P＜0.001），迁移细胞数由（288.40±4.39）个上升至（489.20±5.58）个和（490.40±7.17）个（均P＜0.001）；RKO细胞G0/G1期比例由阴性对照组的（61.17±0.41）%下降至shKIF26A#1组和shKIF26A#2组的（57.13±0.23）%和（57.70±0.06）%（均P＜0.001），侵袭细胞数由（57.60±4.11）个上升至（160.80±5.48）个和（136.20±4.44）个（均P＜0.001），迁移细胞数由（81.00±2.95）个上升至（270.80±4.42）个和（299.00±3.63）个（均P＜0.001）。过表达KIF26A后，HCT116细胞G0/G1期比例由空载体对照组的（41.20±0.88）%上升至KIF26A过表达组的（54.33±0.33）%（P＜0.001），侵袭细胞数由（338.00±5.57）个下降至（194.80±4.90）个（P＜0.001），迁移细胞数由（408.60±3.33）个下降至（263.60±3.59）个（P＜0.001）；RKO细胞G0/G1期比例由空载体对照组的（57.63±0.33）%上升至KIF26A过表达组的（62.65±0.45）%（P＜0.001），侵袭细胞数由（102.60±3.37）个下降至（54.80±2.01）个（P＜0.001），迁移细胞数由（180.00±4.18）个下降至（90.40±4.77）个（P＜0.001）。对TCGA数据库样本KIF26A高表达组和KIF26A低表达组差异表达基因、过表达KIF26A组与空载体对照组RKO细胞差异表达基因的GSEA分析显示，差异表达基因在细胞周期、侵袭和迁移相关通路明显富集（均P＜0.05）。机制研究发现，敲低KIF26A抑制结肠癌细胞中视网膜母细胞瘤蛋白（RB）和E2F转录因子1（E2F1）的表达，促进磷脂酰肌醇3-激酶（PI3K）和AKT丝氨酸/苏氨酸激酶1（AKT1）的表达，而KIF26A过表达则促进RB和E2F1的表达，抑制PI3K和AKT1的表达。Co-IP联合质谱鉴定显示，JunB为KIF26A的相互作用蛋白。ChIP实验结果显示，JunB结合于PTEN的启动子区。回补实验显示，JunB敲低可部分逆转KIF26A高表达所介导的侵袭和迁移能力抑制（P＜0.001）。结论： KIF26A调控RB/E2F1通路影响细胞周期进程，通过与JunB相互作用调控PI3K/AKT信号通路，进而抑制结直肠癌细胞增殖、侵袭和迁移。KIF26A/JunB/PI3K/AKT调控轴的揭示为理解结直肠癌的转移机制提供了新视角，KIF26A有望成为结直肠癌潜在的转移预测标志物及治疗靶点。.