大鼠增殖抑制基因过表达对C6胶质瘤细胞侵袭的抑制作用及其机制探讨

发布时间：2018-03-08 08:25

本文选题：神经胶质瘤　切入点：基因表达调控　出处：《肿瘤》2017年02期 　论文类型：期刊论文

【摘要】：目的 :研究大鼠增殖抑制基因(rat hyperplasia suppressor gene,r HSG)过表达对胶质瘤C6细胞侵袭的影响,并探讨其可能的作用机制。方法 :用携带r HSG基因的重组腺病毒Adv-r HSG-GFP感染胶质瘤C6细胞,同时设置Adv-GFP感染的阴性对照组以及仅PBS处理的空白对照组。采用实时荧光定量PCR和蛋白质印迹法检测各组细胞中r HSG的表达变化。然后,采用细胞划痕愈合实验和Transwell迁移及侵袭实验分别检测r HSG过表达对胶质瘤C6细胞迁移和侵袭的影响,细胞免疫荧光法和细胞黏附实验分别分析r HSG过表达后C6细胞骨架形成和黏附能力的变化,实时荧光定量PCR和蛋白质印迹法测定各组细胞中Rho/Rock信号通路关键分子Ras同源基因家族成员A(Ras homolog gene family,member A,RhoA)、Ras相关的C3肉毒底物1(Ras-related C3 botulinum toxin substrate 1,Rac1)、细胞分裂周期蛋白42(cell division cycle 42,Cdc42)、Rho相关卷曲螺旋形成蛋白激酶1(Rho associated coiled-coil forming protein kinase 1,Rock1)和Rock2的表达水平。结果 :重组腺病毒Adv-r HSG-GFP感染48和72 h后,胶质瘤C6细胞中r HSG的表达水平明显升高(P值均0.01)。与2个对照组相比,Advr HSG-GFP组细胞的划痕愈合率明显降低(P值均0.01),迁移、侵袭和黏附细胞数均明显减少(P值均0.05)。r HSG过表达后C6细胞形态发生变化,丝状伪足缩短,部分呈片状;而且细胞中Rho A、Rac1、Cdc42、Rock1和Rock2的m RNA及蛋白表达水平均明显降低(P值均0.05)。结论 :r HSG过表达可以抑制胶质瘤C6细胞的侵袭,其作用机制可能与阻滞Rho/Rock信号通路有关。
[Abstract]:Aim: to study the effect of rat hyperplasia suppressor gene HSG overexpression on the invasion of glioma C6 cells and its possible mechanism. Methods: glioma C6 cells were infected with Adv-r HSG-GFP, a recombinant adenovirus carrying r HSG gene. At the same time, the negative control group of Adv-GFP infection and the blank control group treated only with PBS were set up. The expression of r HSG was detected by real-time fluorescence quantitative PCR and Western blotting. The effects of r HSG overexpression on the migration and invasion of glioma C6 cells were detected by scratch healing assay and Transwell migration and invasion assay, respectively. The changes of cytoskeleton formation and adhesion ability of C6 cells after r HSG overexpression were analyzed by cell immunofluorescence assay and cell adhesion assay, respectively. Real-time fluorescence quantitative PCR and Western blotting assay for the detection of Ras homolog gene member of Ras homologous gene family member of Rho/Rock signaling pathway in each group of cells, including Ras C 3 botulinum toxin substrate 1 Rac1a, division cycle 42Cdc42C42rho, a member of the Ras homologous gene family member, member of the Ras homolog gene family member, Ras-related C 3 botulinum toxin substrate 1, Rho, and the cell cycle protein 42T cell, division cycle 42Cdc42a, a member of the Ras homologous gene family. Results: the expression levels of Rho associated coiled-coil forming protein kinase 1Rock1) and Rock2 were detected. Results: the recombinant adenovirus Adv-r HSG-GFP was infected 48 and 72 hours after infection. The expression level of r HSG in C6 glioma cells was significantly increased, P value was 0.01 and P value was 0.01, compared with two control groups, the rate of scar healing was significantly decreased and migration was also observed in the Avr HSG-GFP group, and the expression level of r HSG in C6 glioma cells was significantly higher than that in the control group. The number of invasiveness and adhesion cells decreased significantly (P < 0.05). After overexpression of HSG, the morphology of C6 cells was changed, the filamentous pseudopodia was shortened, and some of them were flaky. Moreover, the expression levels of m RNA and protein in Rho AroRac1CDc42C42Rock1 and Rock2 were significantly decreased by 0.050.Conclusion the overexpression of Rho HSG can inhibit the invasion of glioma C6 cells, and its mechanism may be related to blocking the Rho/Rock signaling pathway.
【作者单位】：宁夏医科大学临床医学院;宁夏医科大学宁夏颅脑疾病重点实验室;宁夏医科大学总医院神经外科;榆林市第一医院神经外科;滨州市人民医院神经外科;徐州市肿瘤医院神经外科;
【分类号】：R739.41

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