欢迎您！东篱公司
退出
-
申报数据库
-
立项数据库
-
成果数据库
-
项目获奖数据库
中文摘要：
目的观察抑制胶质瘤细胞LN229中水通道蛋白4（AQP4）的表达对胶质瘤细胞凋亡的影响，探讨其分子机制。方法利用小RNA干扰（siRNA）技术稳定转染恶性胶质瘤细胞株LN229并得到细胞克隆（实验组siAQP4／LN229和对照组scr／LN229），Westernblot技术检测AQP4的蛋白表达；噻唑蓝（MTr）比色法检测细胞的存活率；流式细胞术（FCM）检测细胞色素C（Cyt—C）含量的变化．并用Westernblot检测Cyt—C、bcl-2及Bad的变化。建立裸鼠皮下成瘤模型，接种siAQP4／LN229和ser／LN229两组细胞（每组20只），观察AQP4降表达对移植瘤生长的影响。结果稳定转染LN229筛选出AQP4降表达的稳定克隆（clone2），与对照组比较AQP4的表达明显降低，MTr比色法分别检测在6d内两组细胞的存活率，第6天存活率分别为：scr／LN229组（587．00±4．68）％，siAQP4／N229组（317．00±26．30）％，差异有统计学意义（P〈0．05）；应用流式细胞术检测发现Cyt-C的平均荧光强度分别为scr／LN229组（57．20±16．64），siAQP4／LN229组（468．80±46．90），差异有统计学意义（P〈0．05）；在蛋白水平上，siAQP4／LN229组Cyt—C含量的相对灰度值（1．62±0．16）较scr／LN229组（0．83±0．29）显著升高，siAQP4／LN229组Bad含量（1．30±0．24）较scr／LN229组（0．56±0．21）显著增加，而siAQP4／LN229组bcl-2含量（0．53±0．03）较scr／LN229组（0．73±0．12）的表达明显降低（P〈0．05）。裸鼠皮下成瘤实验显示第6周时，成瘤平均体积分别为对照组（402．67±34．27）mm3，实验组（65．15±32．12）mm。，差异有统计学意义（P〈0．05）。结论AQP4能够调控胶质瘤细胞LN229的凋亡，其调节机制可能与Cyt—C的释放及凋亡相关基因Bad和bcl-2有关。
英文摘要：
Objective To investigate the effects of apuaporin 4 （AQP4） on apoposis of LN229 glioblastoma cells by AQP4 small RNA interference （siRNA） technology and the molecular mechanism. Methods AQP4 expression was knocked down in LN229 glioblastoma cells by AQP4 siRNA （scr/LN229 and siAQP4/LN229 cells）. The expression level of AQP4 protein in LN229 cells was detected by Western blotting. Stable clones were used to measure cells survival ratio （6 days ） by methyl thiazol tetrazolium （MTr） assay. Flow cytometry （FCM） was used to examine the content of cytochrome C in siAQP4/LN229 and scr/LN229. Western blotting was used to measure the levels of cytoehrome C, bcl-2 and Bad. Nu/Nu mice were subcutaneously injected with siAQP4/LN229 and scr/LN229 cells to study the growth of tumor in the two groups （20 mice in each group）. Results Western blotting showed that the expression of AQP4 was decreased significantly; MTT assay suggested that the survival ratio was decreased with deficiency of AQP4： the survival ratio of scr/LN229 was （587. 00± 4. 68 ）%, and the ratio of siAQP4/N229 was （317.00 ± 26.30）% at sixth day （P〈 0.05）. The mean fluorescence density of cytochrome C in scr/LN229 and siAQP4/LN229 by FCM was （ 57.2 ± 16. 64 ） and （ 468.8 ± 46. 9 ） respectively （ P 〈 0. 05 ）. The relative intensity of eytochrome C and Bad in siAQP4/N229 cells was （ 1.62 ± 0. 16 ） and （1.30 ±0. 24） respective/y, and that in scr/LN229 cells was （0. 83 ±0. 29） and （0. 56 ±0. 21） respectively （ P 〈 0. 05 ）. The intensity of bcl-2 in siAQP4/N229 cells （ 0. 53 ± 0. 03 ） was decreased compared to scr/LN229 cells （0. 73 ±0. 12） （P 〈0. 05）. The subcutaneous mouse xenograft model showed that the mean tumor volume of scr/LN229 group was （402. 67 ± 34. 27 ） mm3 , and that of siAQP4/N229 group was （65. 15 ± 32. 12 ） mm3 at the 6th week. Conclusion AQP4 can regulate apoptosis of glioblastoma cell line LN229, which may be related with the release of
位置：
同期刊论文项目
