敲低DCLK1抑制神经胶质瘤细胞增殖、迁移与侵袭

余杰; 杨锋; 邓健; 胡林旺

doi:10.3971/j.issn.1000-8578.2020.20.0127

敲低DCLK1抑制神经胶质瘤细胞增殖、迁移与侵袭

Knockdown of DCLK1 Inhibits Proliferation, Migration and Invasion of Glioma Cells

摘要

摘要:
目的探讨双肾上腺皮质激素样激酶1(DCLK1)对胶质瘤细胞增殖、迁移与侵袭的影响及其机制。
方法 RT-qPCR、免疫组织化学染色和Western blot检测胶质瘤组织和细胞(U87和A172)中DCLK1 mRNA和蛋白的表达情况; sh-DCLK1和阴性对照(sh-Con)转染U87和A172细胞; MTT和Transwell法分析敲低DCLK1对神经胶质瘤细胞增殖、迁移和侵袭能力的影响; 蛋白印迹法检测敲低DCLK1对神经胶质瘤细胞中TGF-β/Smads信号通路相关蛋白TGF-β1、p-Smad2、p-Smad7表达的影响; 将已转染sh-DCLK1或sh-Con的U87细胞注射到BALB/c裸鼠颈部皮下，定期测量瘤体体积，收集瘤体并称重。
结果胶质瘤组织和细胞中DCLK1 mRNA和蛋白表达较癌旁正常组织和正常神经胶质细胞均显著升高(P < 0.001)。与sh-Con组比，sh-DCLK1组的胶质瘤细胞增殖、迁移和侵袭能力及细胞中TGF-β1、p-Smad2和p-Smad7表达显著降低(P < 0.01; P < 0.001);sh-DCLK1组裸鼠体内的瘤体体积和瘤体质量明显降低(P < 0.001)。
结论敲低DCLK1能抑制胶质瘤细胞增殖、迁移和侵袭，其机制可能与抑制TGF-β/Smads信号活性有关。

Abstract:
Objective To investigate the effects of DCLK1 on the proliferation, migration and invasion of glioma cells and their mechanisms.
Methods The mRNA and protein expression of DCLK1 in glioma tissues and glioma cell lines U87 and A172 were detected by RT-qPCR, immunohistochemical staining and Western blot. sh-DCLK1 and the corresponding negative control (sh-Con) were transfected into glioma cell lines U87 and A172 by Lipofectamine 2000 to silence DCLK1 expression. The effects of DCLK1 knockdown on the proliferation, migration and invasion of glioma cells were analyzed by MTT assay and Transwell assay. The effects of DCLK1 knockdown on the expression of TGF-β/Smads signaling pathway-related proteins TGF-β1, p-Smad2 and p-Smad7 in glioma cells were detected by Western blot. U87 cells transfected with sh-DCLK1 or sh-Con were injected into the subcutaneous neck of BALB/c nude mice, the tumor volume and weight were measured.
Results Compared with normal adjacent tissues and normal glial cells, the mRNA and protein expression levels of DCLK1 were significantly increased in glioma tissues and cells (P < 0.001). Compared with sh-Con group, the proliferation, migration and invasion abilities, and the expressions of TGF-β1, p-Smad2 and p-Smad7 proteins of glioma cells in sh-DCLK1 group were significantly decreased (P < 0.01; P < 0.001); in addition, tumor volume and weight of sh-DCLK1 group were significantly reduced in vivo (P < 0.001).
Conclusion DCLK1 knockdown could inhibit the proliferation, migration and invasion of glioma cells, which may be related to the inhibition of TGF-β/Smads signal activity.

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