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. 2019 Mar 7:24:19.
doi: 10.1186/s11658-019-0139-z. eCollection 2019.

Downregulation of CDKL1 suppresses neuroblastoma cell proliferation, migration and invasion

Weiyi Li  1 , Jing Cao  2 , Jian Liu  1 , Wenli Chu  1 , Congqing Zhang  1 , Shuiling Chen  1 , Zefeng Kang  1
Affiliations

Downregulation of CDKL1 suppresses neuroblastoma cell proliferation, migration and invasion

Weiyi Li et al. Cell Mol Biol Lett. .

Retraction in

Abstract

Background: Cyclin-dependent kinase-like 1 (CDKL1) is a member of the cell division control protein 2-related serine-threonine protein kinase family. It is known to occur in various malignant tumors, but its role in neuroblastoma (NB) remains unclear.

Methods: We constructed a CDKL1-silenced NB cell strain (SH-SY5Y) and used real-time PCR and western blotting to confirm the silencing. Functional analyses were performed using the MTT, colony-formation, FACS, wound-healing and transwell invasion assays.

Results: The expression of CDKL1 was significantly upregulated in NB tissue as compared to the adjacent normal tissue. CDKL1 knockdown significantly suppressed cell viability and colony formation ability. It also induced cell cycle G0/G1 phase arrest and apoptosis, and suppressed the migration and invasion ability of SH-SY5Y cells. CDKL1 knockdown decreased the CDK4, cyclin D1 and vimentin expression levels, and increased the caspase-3, PARP and E-cadherin expression levels in SH-SY5Y cells.

Conclusions: Our findings suggest that CDKL1 plays an important role in NB cell proliferation, migration and invasion. It might serve as a potential target for NB therapy.

Keywords: CDKL1; Invasion; Migration; Neuroblastoma; Proliferation; SH-SY5Y.

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Conflict of interest statement

All the clinical specimens were collected in accordance to the Declaration of Helsinki. This study was approved by the Research Ethics Committee of the Eye Hospital of the China Academy of Chinese Medical Sciences (EHCAC-20170635, date: 2017-6-12).Not applicable.The authors declare that they have no competing interests.Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Figures

Fig. 1
Fig. 1
CDKL1 is upregulated in NB tissues. a Real-time PCR analysis of CDKL1 expression in human NB tissues. b Western blotting analysis of CDKL1 expression in human NB tissues. Data are presented as means ± SD. β-actin and GAPDH were used as the internal control. T indicates NB tissues, N indicates normal nerve tissues
Fig. 2
Fig. 2
The efficiency of CDKL1 using an siRNA lentivirus. a Western blotting analysis of the CDKL1 expression in human NB cell lines. b Lentivirus infection in the SH-SY5Y cell line. Bright and fluorescent photomicrographs of SH-SY5Y cells were taken 48 h after lentivirus infection. c and d Real-time PCR and western blotting were used to determine the CDKL1 expression in SH-SY5Y cells infected with siCDKL1 or the negative control (NC) siRNA lentivirus. Data are presented as means ± SD. **p < 0.01 vs. NC
Fig. 3
Fig. 3
The effect of CDKL1 knockdown on the proliferation, cell cycle distribution and apoptosis of NB cells in vitro. a Analysis of cell viability of siCDKL1-transfected cells based on the MTT assay. b Photomicrographs and statistical analyses of colonies of SH-SY5Y cells after transfection with siCDKL1 or NC. c The effect of CDKL1 knockdown on the cell cycle profile of SH-SY5Y cells by FACS. The percentage of cells in G0/G1, S and G2/M phases are shown. d Lentivirus-infected SH-SY5Y cells were analyzed using flow cytometry analysis for positive annexin V–APC and 7-AAD staining . Data are presented as means ± SD. **p < 0.01, ***p < 0.001 vs. NC
Fig. 4
Fig. 4
The effect of CDKL1 knockdown on cell migration and invasion of NB cells in vitro. a Wound-healing assays were performed to investigate the migratory ability of SH-SY5Y cells. b Representative images are shown for siCDKL1-transfected SH-SY5Y cells in a transwell assay. The invasion was quantified by counting the cells in 3 random microscopic fields. Data are presented as means ± SD. **P < 0.01, ***P < 0.001 vs. NC
Fig. 5
Fig. 5
Effect of CDKL1 knockdown on the regulation of cell cycle, apoptotic and invasion in SH-SY5Y cells, as confirmed using western blot analysis. GAPDH was used as the internal control. Data are presented as means ± SD. **p < 0.01, ***p < 0.001 vs. NC
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