Wu Ke-Jun, Zeng Da-Tong, He Rong-Quan, Li Dong-Ming, Yao Jin-Lian, Liu Li-Min, Huang Wei-Jian, Qin Di-Yuan, Li Yu-Feng, He Han, Li Shi-De, Wen Jia-Ying, Meng Li, Shi Jia-Rong, Chen Gang, Li Hui
Department of Pathology, The First Affiliated Hospital of Guangxi Medical University, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China.
Department of Medical Oncology, The First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China.
World J Clin Oncol. 2025 Jul 24;16(7):108666. doi: 10.5306/wjco.v16.i7.108666.
The prevalence of colorectal cancer (CRC) in younger people is increasing. Despite advances in precision medicine, the challenges of drug resistance and high costs persist. Nitidine chloride (NC) has pharmacological potential, and kinesin family member 20A (KIF20A) is overexpressed in various tumors; however, their interaction in CRC remains unexplored.
To investigate the KIF20A expression characteristics in CRC cells and determine whether it is a potential target gene for NC in inhibiting CRC treatment.
Single-cell RNA sequencing (scRNA-seq), spatial transcriptomics, and mRNA expression profiling were used to analyze KIF20A expression in CRC cells. Immunohistochemical staining was used to verify KIF20A expression in 416 clinical samples (208 CRC tissue samples and 208 noncancerous control tissue samples). Clustered regularly interspaced short palindromic repeats (CRISPR) technology was used to evaluate the impact of knocking out KIF20A on CRC cell growth. Molecular docking was applied to analyze NC-KIF20A binding. Finally, RNA sequencing and functional enrichment analysis were performed to explore the mechanism of action of NC in CRC cells.
Treating HCT116 cells with NC was found to significantly downregulate KIF20A ( < 0.05), and the molecular docking analysis revealed high-affinity binding between NC and KIF20A (binding energy = -9.6 kcal/mol). The scRNA-seq, spatial transcriptomics, and mRNA expression profiling results confirmed the significantly high expression of KIF20A in CRC tissues (standardized mean difference = 1.33, 95% confidence interval: 0.885-1.77, summary receiver operating characteristic curve area = 0.94). The immunohistochemical analysis of the clinical samples showed high KIF20A expression in the CRC tissues ( < 0.05), with significant correlation between the level of expression and gender, tumor size, and tumor grade ( < 0.05). Knocking out KIF20A significantly inhibited the growth of various CRC cell lines (CRISPR score < -0.3). The functional enrichment analysis indicated that NC may inhibit CRC by disrupting several biological processes, such as mitotic nuclear division, chromosome segregation, and microtubule binding.
Our results indicate that NC binds to KIF20A with high affinity and downregulates its expression in CRC cells, leading to reduced proliferation. Hence, NC has promise as a therapeutic agent in the treatment of CRC, and targeting KIF20A also has potential as a therapeutic strategy. Further KIF20A knockout studies are needed to confirm the binding specificity and mechanistic roles of NC in CRC.
结直肠癌（CRC）在年轻人中的患病率正在上升。尽管精准医学取得了进展，但耐药性和高成本等挑战仍然存在。氯化两面针碱（NC）具有药理潜力，驱动蛋白家族成员20A（KIF20A）在多种肿瘤中过表达；然而，它们在结直肠癌中的相互作用仍未得到探索。
研究KIF20A在结直肠癌细胞中的表达特征，并确定其是否为NC抑制结直肠癌治疗的潜在靶基因。
采用单细胞RNA测序（scRNA-seq）、空间转录组学和mRNA表达谱分析KIF20A在结直肠癌细胞中的表达。免疫组织化学染色用于验证416例临床样本（208例结直肠癌组织样本和208例非癌对照组织样本）中KIF20A的表达。利用成簇规律间隔短回文重复序列（CRISPR）技术评估敲除KIF20A对结直肠癌细胞生长的影响。应用分子对接分析NC与KIF20A的结合情况。最后，进行RNA测序和功能富集分析，以探索NC在结直肠癌细胞中的作用机制。
发现用NC处理HCT116细胞可显著下调KIF20A（<0.05），分子对接分析显示NC与KIF20A之间具有高亲和力结合（结合能=-9.6 kcal/mol）。scRNA-seq、空间转录组学和mRNA表达谱结果证实KIF20A在结直肠癌组织中显著高表达（标准化平均差异=1.33，95%置信区间：0.885-1.77，汇总受试者工作特征曲线面积=0.94）。临床样本的免疫组织化学分析显示结直肠癌组织中KIF20A高表达（<0.05），表达水平与性别、肿瘤大小和肿瘤分级之间存在显著相关性（<0.05）。敲除KIF20A显著抑制了各种结直肠癌细胞系的生长（CRISPR评分<-0.3）。功能富集分析表明，NC可能通过破坏有丝分裂核分裂、染色体分离和微管结合等多种生物学过程来抑制结直肠癌。
我们的结果表明，NC与KIF20A具有高亲和力结合，并下调其在结直肠癌细胞中的表达，导致增殖减少。因此，NC有望成为治疗结直肠癌的治疗药物，靶向KIF20A也具有作为治疗策略的潜力。需要进一步进行KIF20A敲除研究，以确认NC在结直肠癌中的结合特异性和机制作用。
