Qin Chunzhi, Ren Li, Ji Meiling, Lv Shixu, Wei Ye, Zhu Dexiang, Lin Qi, Xu Pingping, Chang Wenju, Xu Jianmin
Department of General Surgery, Zhongshan Hospital, Fudan University, Shanghai.
Department of Surgical Oncology, First Affiliated Hospital of Wenzhou Medical University, Wenzhou, People's Republic of China.
Onco Targets Ther. 2017 Mar 16;10:1613-1624. doi: 10.2147/OTT.S133014. eCollection 2017.
CDKL1 is a member of the cell division cycle 2 (CDC2)-related serine threonine protein kinase family and is overexpressed in malignant tumors such as melanoma, breast cancer, and gastric cancer.
This study aimed to evaluate whether CDKL1 can serve as a potential molecular target for colorectal cancer therapy.
Expression of CDKL1 in colorectal cancer tissues and cell lines was measured by immunohistochemistry and Western blot, respectively. To investigate the role of CDKL1 in colorectal cancer, CDKL1-small hairpin RNA-expressing lentivirus was constructed and infected into HCT116 and Caco2 cells. The effects of RNA interference (RNAi)-mediated CDKL1 downregulation on cell proliferation and invasion were assessed by CCK-8, colony formation, transwell, and tumorigenicity assays in nude mice. The effects of CDKL1 downregulation on cell cycle and apoptosis were analyzed by flow cytometry. Furthermore, microarray method and data analysis elucidated the molecular mechanisms underlying the phenomenon.
CDKL1 protein was overexpressed in colorectal cancer tissues compared with paired normal tissues. Knockdown of CDKL1 in HCT116 and Caco2 significantly inhibited cell growth, colony formation ability, tumor invasion, and G1-S phase transition of the cell cycle. The knockdown of stimulated the upregulation of p15 and retinoblastoma protein.
CDKL1 plays a vital role in tumor proliferation and invasion in colorectal cancer in vitro and in vivo and, thus, may be considered as a valuable target for therapeutic intervention.
细胞周期蛋白依赖性激酶样蛋白1（CDKL1）是与细胞分裂周期2（CDC2）相关的丝氨酸/苏氨酸蛋白激酶家族成员，在黑色素瘤、乳腺癌和胃癌等恶性肿瘤中过表达。
本研究旨在评估CDKL1是否可作为结直肠癌治疗的潜在分子靶点。
分别采用免疫组织化学和蛋白质免疫印迹法检测CDKL1在结直肠癌组织及细胞系中的表达。为研究CDKL1在结直肠癌中的作用，构建了表达CDKL1短发夹RNA的慢病毒，并将其感染HCT116和Caco2细胞。通过CCK-8法、集落形成实验、Transwell实验及裸鼠成瘤实验评估RNA干扰（RNAi）介导的CDKL1下调对细胞增殖和侵袭的影响。采用流式细胞术分析CDKL1下调对细胞周期和凋亡的影响。此外，通过基因芯片方法及数据分析阐明该现象的分子机制。
与配对的正常组织相比，CDKL1蛋白在结直肠癌组织中过表达。在HCT116和Caco2细胞中敲低CDKL1可显著抑制细胞生长、集落形成能力、肿瘤侵袭及细胞周期的G1-S期转变。CDKL1敲低可刺激p15和视网膜母细胞瘤蛋白上调。
CDKL1在体外和体内的结直肠癌肿瘤增殖和侵袭中起重要作用，因此，可被视为治疗干预的有价值靶点。
