RNAi-mediated downregulation of CDKL1 inhibits growth and colony-formation ability, promotes apoptosis of human melanoma cells.
BACKGROUND
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.


OBJECTIVE
This study aimed to evaluate the whether CDKL1 can serve as a potential molecular target for melanoma gene therapy.


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Department of Dermatology, The 1st Affiliated Hospital of Dalian Medical University, No. 222, Zhongshan Road, Dalian 116011, China.
Zhiqi
Z
Song
Department of Dermatology, The 1st Affiliated Hospital of Dalian Medical University, No. 222, Zhongshan Road, Dalian 116011, China. Electronic address: amy4963@163.com.
Jingrong
J
Lin
Department of Dermatology, The 1st Affiliated Hospital of Dalian Medical University, No. 222, Zhongshan Road, Dalian 116011, China.
Zhe
Z
Sun
Department of Dermatology, The 1st Affiliated Hospital of Dalian Medical University, No. 222, Zhongshan Road, Dalian 116011, China.
Jing
J
Ni
Department of Dermatology, The 1st Affiliated Hospital of Dalian Medical University, No. 222, Zhongshan Road, Dalian 116011, China.
Yang
Y
Sha
D017209:Apoptosis; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D003114:Colony-Forming Units Assay; D018844:Cyclin-Dependent Kinases / Q000378:metabolism*; D015536:Down-Regulation; D059585:G1 Phase Cell Cycle Checkpoints; D015870:Gene Expression; D055785:Gene Knockdown Techniques; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus; D008545:Melanoma / Q000235:genetics* / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism*; D034741:RNA, Small Interfering; D012878:Skin Neoplasms / Q000235:genetics* / Q000378:metabolism; D014410:Tumor Stem Cell Assay
D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; C096702:CDKL1 protein, human; D018844:Cyclin-Dependent Kinases
CDKL1; Lentivirus; Melanoma; siRNA