摘要
目的:研究鞘氨醇激酶1(SphK1)表达下调对膀胱癌T24细胞增殖和细胞迁移的影响。方法:SphK1siRNA和无义siRNA分别被转染至膀胱癌T24细胞后将细胞分为对照组、无义siRNA组和SphK1siRNA组。采用Western blotting检测三组细胞中SphK1蛋白的表达,采用CCK-8分析转染SphK1siRNA前后细胞增殖能力的变化,采用Boyden小室检测三组细胞迁移能力的改变,采用Western blotting分析cdk2和p21以及MMP-2和MMP-9蛋白表达。结果:SphK1siRNA组中T24细胞的SphK1蛋白表达低于对照组和无义siRNA组(P<0.05)。与对照组和无义siRNA组相比,SphK1siRNA组中细胞增殖受到抑制(P<0.05),且SphK1siRNA组中p21蛋白表达高于对照组和无义siRNA组,cdk2蛋白的表达降低(P<0.05)。SphK1siRNA组中细胞穿膜数低于对照组和无义siRNA组(P<0.05),且SphK1siRNA能引起MMP-2和MMP-9表达的下调(P<0.05)。结论:SphK1可能在膀胱癌的发生发展中发挥重要作用,其表达下调引发的细胞增殖抑制和细胞迁移降低可能与p21表达上调和cdk2、MMP-2和MMP-9表达下调密切相关。
Objective To investigate the effect of down-regulation of sphingosine kinase 1 (SphK1) on cell proliferation and cell migration of bladder carcinoma T24 cells. Methods SphK1 siRNA and nonsense siRNA were used to transfect bladder carcinoma T24 cells, which was divided into three groups: control group, nonsense siRNA group and SphK1 siRNA group. Expression of SphK1 protein was detected in three group T24 cells by Western blotting method. The changes of cell proliferation were analyzed by CCK-8 kit after transfection with SphK1 siRNA, and the changes of cell migration ability were detected by Boyden chamber. Expressions of cdk2, p21, MMP-2,and MMP-9 proteins were examined by Western blotting method. Results Expression of SphK1 protein in SphK1 siRNA group was lower than those in control group and nonsense siRNA group (P 〈 0.05 ). The result of cell proliferation demonstrated that compared with control group and nonsense siRNA group, cell proliferation in SphK1 siRNA group was obviously inhibited (P 〈 0.05), coupled with the elevation of p21 protein and decrease of cdk2 protein (P 〈 0.05 ). The number of cells migrated through membrane in SphK1 siRNA group was markedly lower than those in control group and nonsense siRNA group (P 〈 0.05), accompanied by the down-regulation of MMP-2 and MMP-9 proteins (P 〈 0.05). Conclusion SphK1 may play an essential role in the occurrence and development of bladder carcinoma, and its down-regulation mediated proliferation inhibition and cell migration decrease may be closely associated with the elevation of p21 expression and the decrease of cdk2, MMP-2 and MMP-9 proteins.
出处
《实用医学杂志》
CAS
北大核心
2013年第8期1218-1221,共4页
The Journal of Practical Medicine