Wang Penghui, He Xiaodong
Department of General Surgery, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College Beijing, China.
Department of General Surgery, Beijing Tiantan Hospital, Capital Medical University Beijing, China.
Int J Clin Exp Pathol. 2020 May 1;13(5):923-933. eCollection 2020.
Cytoskeleton-associated protein 2-like (CKAP2L) exerts crucial function in the cell-cycle progression and mitotic spindle formation of neural stem/progenitor cells. However, in hepatocellular carcinoma (HCC), the expression pattern, clinical significance and biologic role of CKAP2L remain unexplored. We analysed The Cancer Genome Atlas (TCGA) database and found that CKAP2L was dramatically upregulated in HCC tissues at the mRNA level compared to adjacent tissues, which was validated in 48 paired HCC and para-tumor tissues using quantitative real-time PCR (qRT-PCR). Immunohistochemical analysis of tissue microarray revealed that CKAP2L was also significantly overexpressed at the protein level. Further clinical and survival analysis of the TCGA cohort revealed that increased CKAP2L expression was strongly associated with reduced overall survival. We further validated that higher CKAP2L protein expression was associated with worse prognosis in the Peking Union Medical College Hospital (PUMCH) cohort. Univariate and multivariate Cox regression analyses in the TCGA and PUMCH cohort suggested that CKAP2L overexpression was an independent risk factor for poor prognosis in HCC patients. Then, we validated that CKAP2L silencing inhibited HCC cell proliferation, migration, and invasion abilities. Knockdown of CKAP2L in Huh7 cells suppressed the growth of xenograft tumors in vivo. Furthermore, qRT-PCR and western blotting results demonstrated that the expression of Class I Phosphoinositide 3-Kinase PIK3CA/p110α and PIK3CB/p110β isoforms reduced obviously in Huh7 cells after depleting CKAP2L. This study demonstrated for the first time that high CKAP2L expression in HCC tissues is significantly correlated with poor prognosis in HCC patients and greatly facilitate the malignancy of HCC, thus providing a new prognostic biomarker and potential therapeutic target.
细胞骨架相关蛋白2样蛋白（CKAP2L）在神经干/祖细胞的细胞周期进程和有丝分裂纺锤体形成中发挥关键作用。然而，在肝细胞癌（HCC）中，CKAP2L的表达模式、临床意义和生物学作用仍未得到探索。我们分析了癌症基因组图谱（TCGA）数据库，发现与邻近组织相比，HCC组织中CKAP2L在mRNA水平上显著上调，这在48对HCC和癌旁组织中通过定量实时PCR（qRT-PCR）得到了验证。组织芯片的免疫组织化学分析显示，CKAP2L在蛋白水平上也显著过表达。对TCGA队列的进一步临床和生存分析表明，CKAP2L表达增加与总生存期缩短密切相关。我们进一步验证了在北京协和医院（PUMCH）队列中，较高的CKAP2L蛋白表达与较差的预后相关。在TCGA和PUMCH队列中的单因素和多因素Cox回归分析表明，CKAP2L过表达是HCC患者预后不良的独立危险因素。然后，我们验证了CKAP2L沉默可抑制HCC细胞的增殖、迁移和侵袭能力。在Huh7细胞中敲低CKAP2L可抑制体内异种移植肿瘤的生长。此外，qRT-PCR和蛋白质印迹结果表明，在耗尽CKAP2L后，Huh7细胞中I类磷脂酰肌醇3激酶PIK3CA/p110α和PIK3CB/p110β亚型的表达明显降低。本研究首次表明，HCC组织中高CKAP2L表达与HCC患者的不良预后显著相关，并极大地促进了HCC的恶性程度，从而提供了一种新的预后生物标志物和潜在的治疗靶点。
