TRPV2基因敲除对胶质瘤U87MG细胞增殖能力的影响  

The Effect of TRPV2 Knocking-Out on U87MG Cells Proliferation in vitro

作　　者：胡争[1] 芮路[2] 何丹[3] 

机构地区：[1]华中科技大学同济医学院附属同济医院妇产科,武汉430000 [2]阜外心血管病医院心外科,北京100037 [3]华中科技大学同济医学院附属同济医院神经内科,武汉430000

出　　处：《神经损伤与功能重建》2012年第2期84-87,共4页Neural Injury and Functional Reconstruction

摘　　要：目的:探讨瞬时感受器电位离子通道蛋白V亚族2(TRPV2)基因敲除对胶质瘤U87MG细胞增殖及细胞周期进展的影响。方法:通过锌指核酶敲除U87MG细胞的TRPV2基因;Western Blot测定TRPV2蛋白的表达;Brdu掺入实验及Western Blot测定TRPV2基因敲除对U87MG细胞增殖的影响;流式细胞技术测定TRPV2基因敲除对U87MG细胞细胞周期进展的影响。结果:TRPV2蛋白在U87MG细胞中转录表达;靶向TRPV2的锌指核酶能有效地阻断U87MG细胞内TRPV2基因在蛋白水平上的表达。TRV2-/-U87MG细胞Brdu阳性率为(47.37±4.03)%,处于S期细胞的百分率为(20.14±0.47)%,均显著高于TRV2+/+U87MG细胞(P<0.01);PCNA表达量为U87MG TRV2+/+细胞的(1.47±0.19)倍(P<0.01)。结论:TRPV2基因敲除能明显促进U87MG细胞的增殖和细胞周期的进展。Objective： To observe the effect of TRPV2 knocking-out on cultured U87MG cell＇s proliferation in vitro.Methods： The TRPV2 gene of U87MG cells was knocked-out by zinc finger nuclease.The expression of TRPV2 was detected by western blot,and the effect of TRPV2 knocking-out on cultured U87MG cell＇s proliferation and cell cycle progression were measured by Brdu incorporation assay,western blot and flow cytometry respectively.Results： TRPV2 protein expressed on U87MG cells and the expression of TRPV2 protein were decreased by zinc finger nuclease.In TRV2-/-U87MG cells,the percentage of Brdu（＋） cells was（47.37±4.03）% and the percentage of cells in S phase was（20.14±0.47）%,significantly higher than those in TRV2＋/＋ U87MG cells（P0.01）.The expression of PCNA protein in TRV2-/-U87MG cells was（1.47±0.19） fold relative to that in TRV2＋/＋ U87MG cells（P0.01）.Conclusion： TRPV2 knocking-out can significantly promote U87MG cells proliferation and cell cycle progression in vitro.

关 键 词：瞬时感受器电位离子通道蛋白V亚族2 锌指核酶 U87-MG细胞 细胞增殖 细胞周期 

分 类 号：R741[医药卫生—神经病学与精神病学] Q813[医药卫生—临床医学]