目的探究β2整合素/FLNa相互作用的力学调控机制、磷酸化效应与分子结构基础。方法β2整合素/FLNa-WT和T758P晶体结构取自蛋白质数据库(protein data bank,PDB),进行分子动力学(molecular dynamics,MD)模拟,采用MM/PBSA方法计算复合物...目的探究β2整合素/FLNa相互作用的力学调控机制、磷酸化效应与分子结构基础。方法β2整合素/FLNa-WT和T758P晶体结构取自蛋白质数据库(protein data bank,PDB),进行分子动力学(molecular dynamics,MD)模拟,采用MM/PBSA方法计算复合物结合自由能变化,并分析构象演化与残基相互作用等数据。结果β2整合素THR758磷酸化修饰后,复合物的结合自由能下降,拉力累积降低。单纯的力学信号刺激下,β2整合素/FLNa复合物的解离呈现出双相力依赖特性,而磷酸化后复合物的结合解离过程存在单纯的滑移键机制。结论磷酸化将通过减弱M762-G2269残基相互作用,下调β2整合素/FLNa复合物的结合亲和力,张力将双相调节复合物的解离。研究结果有助于加深对炎症反应过程的认识,并为相关药物靶点的发现和抗体设计提供有益参考。展开更多
AIM:To determine the expression and significance of filamin A(FLNa)in colorectal adenocarcinoma tissue.METHODS:The expression of FLNa in 46 colorectal cancer tissues and normal tissues was detected by immunohistochemi...AIM:To determine the expression and significance of filamin A(FLNa)in colorectal adenocarcinoma tissue.METHODS:The expression of FLNa in 46 colorectal cancer tissues and normal tissues was detected by immunohistochemistry,reverse transcription polymerase chain reaction(RT-PCR)and Western blotting,and its relationship with clinical parameters and prognosis was analyzed.RESULTS:The positive expression of FLNa in cancer tissues was lower than that in normal mucosa,and the difference was statistically significant.The expression of FLNa correlated with liver metastasis,lymph node metastasis and rectal invasion depth,regardless of sex,age,tumor location,tumor size,gross shape and histological type of colorectal carcinoma.Multivariate analysis showed that FLNa was an independent risk factor for postoperative survival of patients with colorectal adenocarcinoma.Moreover,survival analysis showed that the expression level of FLNa was closely related with survival of patients with colorectal adenocarcinoma.The results of RT-PCR and Western blotting were consistent with those of immunohistochemistry.CONCLUSION:FLNa showed low expression in colorectal adenocarcinoma,high correlation with the incidence and development of colorectal cancer,and was considered an indicator of prognosis.展开更多
Filamin A and 14-3-3-σ are closely associated with the development of breast cancer. However, the exact relationship between them is still unknown. The present study aimed to examine the interaction of filamin A with...Filamin A and 14-3-3-σ are closely associated with the development of breast cancer. However, the exact relationship between them is still unknown. The present study aimed to examine the interaction of filamin A with 14-3-3-σ in the invasion and migration of breast cancer. RNA interference technology was employed to silence filamin A in MDA-MB-231 cells. Real-time PCR and Westem blotting were used to detect the expression of filamin A and 14-3-3-σ at mRNA and protein levels, respectively. Double immunofluorescence was applied to show their colocalization morphologically. Wound healing assay and Trans-well assay were used to testify the migration and invasion of MDA-MB-231 cells in filamin A-silenced cells. The results showed that silencing filamin A significantly increased the mRNA and protein levels of 14-3-3σ. In addition, double immunofluorescence displayed that filamin A and 14-3-3σ were predominantly colocalized in the cytoplasm of MDA-MB-231 cells. Silencing filamin A led to the enhanced fluorescence of 14-3-3σ. Furthermore, cell functional experiments showed that silencing filamin A inhibited the migration and invasion of MDA-MB-231 cells in vitro. In conclusion, silencing filamin A may inhibit the invasion and migration of breast cancer cells by upregulating 14-3-3σ.展开更多
文摘目的探究β2整合素/FLNa相互作用的力学调控机制、磷酸化效应与分子结构基础。方法β2整合素/FLNa-WT和T758P晶体结构取自蛋白质数据库(protein data bank,PDB),进行分子动力学(molecular dynamics,MD)模拟,采用MM/PBSA方法计算复合物结合自由能变化,并分析构象演化与残基相互作用等数据。结果β2整合素THR758磷酸化修饰后,复合物的结合自由能下降,拉力累积降低。单纯的力学信号刺激下,β2整合素/FLNa复合物的解离呈现出双相力依赖特性,而磷酸化后复合物的结合解离过程存在单纯的滑移键机制。结论磷酸化将通过减弱M762-G2269残基相互作用,下调β2整合素/FLNa复合物的结合亲和力,张力将双相调节复合物的解离。研究结果有助于加深对炎症反应过程的认识,并为相关药物靶点的发现和抗体设计提供有益参考。
文摘目的研究胃癌组织中细丝蛋白A (filamin A, FLNa)、血管内皮生长因子(vascular endothelial growth factor,VEGF)的表达水平和微血管密度(microvascular density, MVD)的关系,探讨FLNa、 VEGF和MVD之间的相关性及临床意义。方法采用免疫组织化学法检测51例胃癌组织及正常胃黏膜组织中FLNa、 VEGF和CD34的表达情况,结合临床参数分析三者相关性及临床意义。结果胃癌组织中的FLNa阳性表达率低于正常胃黏膜组织[47.06%(24/51) vs 90.20%(46/51),P=0.000];胃癌组织中的VEGF阳性表达率高于正常胃黏膜组织[92.16%(47/51)vs 31.37%(16/51),P=0.000];胃癌组织中的MVD值高于正常胃黏膜组织(18±3 vs 5±1,P=0.001);胃癌组织中FLNa、 VEGF的表达水平及MVD值与患者癌灶的浸润深度、是否存在淋巴结转移、肝转移有关,而与年龄、性别、癌灶部位、组织类型及Borrmann分型无关; FLNa和VEGF在胃癌组织中的表达水平呈负相关(r=-0.490,P=0.000);胃癌组织中FLNa表达阳性组的MVD值低于FLNa表达阴性组(4±1 vs 17±3,P=0.001),而VEGF表达阳性组的MVD值高于VEGF表达阴性组(16±1 vs 3±1,P=0.000)。结论 FLNa、 VEGF的表达水平和MVD值与胃癌的形成和发展相关, FLNa通过抑制VEGF的表达水平,阻遏了肿瘤新生血管的生成,最终抑制胃癌细胞的局部复发和远处转移。
基金Supported by Hebei Province Health Department of China,No.14277710D and No.GL2012050
文摘AIM:To determine the expression and significance of filamin A(FLNa)in colorectal adenocarcinoma tissue.METHODS:The expression of FLNa in 46 colorectal cancer tissues and normal tissues was detected by immunohistochemistry,reverse transcription polymerase chain reaction(RT-PCR)and Western blotting,and its relationship with clinical parameters and prognosis was analyzed.RESULTS:The positive expression of FLNa in cancer tissues was lower than that in normal mucosa,and the difference was statistically significant.The expression of FLNa correlated with liver metastasis,lymph node metastasis and rectal invasion depth,regardless of sex,age,tumor location,tumor size,gross shape and histological type of colorectal carcinoma.Multivariate analysis showed that FLNa was an independent risk factor for postoperative survival of patients with colorectal adenocarcinoma.Moreover,survival analysis showed that the expression level of FLNa was closely related with survival of patients with colorectal adenocarcinoma.The results of RT-PCR and Western blotting were consistent with those of immunohistochemistry.CONCLUSION:FLNa showed low expression in colorectal adenocarcinoma,high correlation with the incidence and development of colorectal cancer,and was considered an indicator of prognosis.
文摘Filamin A and 14-3-3-σ are closely associated with the development of breast cancer. However, the exact relationship between them is still unknown. The present study aimed to examine the interaction of filamin A with 14-3-3-σ in the invasion and migration of breast cancer. RNA interference technology was employed to silence filamin A in MDA-MB-231 cells. Real-time PCR and Westem blotting were used to detect the expression of filamin A and 14-3-3-σ at mRNA and protein levels, respectively. Double immunofluorescence was applied to show their colocalization morphologically. Wound healing assay and Trans-well assay were used to testify the migration and invasion of MDA-MB-231 cells in filamin A-silenced cells. The results showed that silencing filamin A significantly increased the mRNA and protein levels of 14-3-3σ. In addition, double immunofluorescence displayed that filamin A and 14-3-3σ were predominantly colocalized in the cytoplasm of MDA-MB-231 cells. Silencing filamin A led to the enhanced fluorescence of 14-3-3σ. Furthermore, cell functional experiments showed that silencing filamin A inhibited the migration and invasion of MDA-MB-231 cells in vitro. In conclusion, silencing filamin A may inhibit the invasion and migration of breast cancer cells by upregulating 14-3-3σ.
