Song Zhiqi, Lin Jingrong, Sun Zhe, Ni Jing, Sha Yang
Department of Dermatology, The 1st Affiliated Hospital of Dalian Medical University, No. 222, Zhongshan Road, Dalian 116011, China.
Department of Dermatology, The 1st Affiliated Hospital of Dalian Medical University, No. 222, Zhongshan Road, Dalian 116011, China.
J Dermatol Sci. 2015 Jul;79(1):57-63. doi: 10.1016/j.jdermsci.2015.03.020. Epub 2015 Apr 9.
Cyclin-dependent kinase-like 1 (CDKL1) is a member of cell division control protein 2 (CDC-2)-related serine threonine protein kinase family, and is reported to be overexpressed in malignant tumors such as breast cancer and gastric cancer.
This study aimed to evaluate the whether CDKL1 can serve as a potential molecular target for melanoma gene therapy.
CDKL1 expression in two melanoma cell lines, A375 and MV3 was measured by real-time PCR. To investigate the role of CDKL1 in cell growth of melanoma, we constructed CDKL1-siRNA expressing lentivirus and infected A375 and MV3 cells. The effects of RNAi-mediated CDKL1 downregulation on A375 and MV3 cell proliferation and colony-formation ability were detected by methylthiazoletetrazolium (MTT) assay and colony-formation assay. The effects of CDKL1 downregulation on A375 and MV3 cell cycle and apoptosis were analyzed by FACS analysis.
Human melanoma cell lines A375 and MV3 expressed CDKL1 mRNA. Knockdown of CDKL1 in A375 and MV3 by CDKL1-siRNA lentivirus infection significantly inhibited cell growth and colony formation ability, promoted cell apoptosis, and arrested cells in G1 phase.
CDKL1 is associated with melanoma cell growth, colony formation, apoptosis, and cell cycle regulation. It may be considered as a valuable target for anti-melanoma therapeutic strategies.
细胞周期蛋白依赖性激酶样1（CDKL1）是细胞分裂控制蛋白2（CDC-2）相关的丝氨酸苏氨酸蛋白激酶家族成员，据报道在乳腺癌和胃癌等恶性肿瘤中过表达。
本研究旨在评估CDKL1是否可作为黑色素瘤基因治疗的潜在分子靶点。
采用实时荧光定量PCR检测两种黑色素瘤细胞系A375和MV3中CDKL1的表达。为研究CDKL1在黑色素瘤细胞生长中的作用，构建表达CDKL1-siRNA的慢病毒并感染A375和MV3细胞。采用甲基噻唑基四氮唑蓝（MTT）法和集落形成试验检测RNA干扰介导的CDKL1下调对A375和MV3细胞增殖及集落形成能力的影响。通过流式细胞术分析CDKL1下调对A375和MV3细胞周期及凋亡的影响。
人黑色素瘤细胞系A375和MV3表达CDKL1 mRNA。通过CDKL1-siRNA慢病毒感染敲低A375和MV3中的CDKL1可显著抑制细胞生长和集落形成能力，促进细胞凋亡，并使细胞停滞于G1期。
CDKL1与黑色素瘤细胞生长、集落形成、凋亡及细胞周期调控相关。它可能被视为抗黑色素瘤治疗策略的一个有价值的靶点。
