下调 CDKL1 表达抑制神经母细胞瘤细胞的增殖、迁移和侵袭。
Downregulation of CDKL1 suppresses neuroblastoma cell proliferation, migration and invasion.
机构信息
1Eye Hospital, China Academy of Chinese Medical Sciences, No 33 Lugu Road, Shijingshan district, Beijing, 100040 China.
Yinan Branch of Qilu Hospital of Shandong University, Linyi, Shandong China.
出版信息
Cell Mol Biol Lett. 2019 Mar 7;24:19. doi: 10.1186/s11658-019-0139-z. eCollection 2019.
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.
背景
周期素依赖性激酶样 1（CDKL1）是细胞分裂控制蛋白 2 相关丝氨酸-苏氨酸蛋白激酶家族的成员。已知其存在于各种恶性肿瘤中，但在神经母细胞瘤（NB）中的作用尚不清楚。
方法
我们构建了 CDKL1 沉默的 NB 细胞株（SH-SY5Y），并使用实时 PCR 和 Western blot 验证沉默效果。通过 MTT、集落形成、FACS、划痕愈合和 Transwell 侵袭实验进行功能分析。
结果
与相邻正常组织相比，NB 组织中 CDKL1 的表达明显上调。CDKL1 敲低显著抑制细胞活力和集落形成能力。它还诱导细胞周期 G0/G1 期阻滞和细胞凋亡，并抑制 SH-SY5Y 细胞的迁移和侵袭能力。CDKL1 敲低降低了 SH-SY5Y 细胞中 CDK4、cyclin D1 和波形蛋白的表达水平，增加了 caspase-3、PARP 和 E-钙黏蛋白的表达水平。
结论
我们的研究结果表明，CDKL1 在 NB 细胞增殖、迁移和侵袭中发挥重要作用。它可能成为 NB 治疗的潜在靶点。
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