Smo、Gli2及FoxM1对神经胶质瘤诊断和预后的评估价值

目的探讨Smo、胶质瘤相关癌基因同源蛋白2(Gli2)及叉头框转录因子M1(FoxM1)对神经胶质瘤诊断和预后的评估价值。 方法取80例神经胶质瘤标本为神经胶质瘤组,60例正常脑组织标本为正常组。免疫组化法检测两组Smo、Gli2、FoxM1表达水平,分析其与神经胶质瘤患者临床病理特征的关系;Kaplan-Meier生存曲线分析不同Smo、Gli2、FoxM1表达水平神经胶质瘤患者预后的差异,分析影响神经胶质瘤预后的相关因素。 结果神经胶质瘤组织Smo、Gli2、FoxM1高表达率均高于正常脑组织,且随神经胶质瘤患者组织学分级升高而表达率增高(P＜0.05)。不同组织学分级、不同Smo、Gli2、FoxM1表达水平的神经胶质瘤患者5年生存率存在显著差异(P＜0.05)。组织学Ⅲ~Ⅳ级、Smo高表达、Gli2高表达、FoxM1高表达是神经胶质瘤预后的危险因素(P＜0.05)。 结论Smo、Gli2、FoxM1高表达是神经胶质瘤预后的危险因素,三者有望作为神经胶质瘤诊断和预后的判断指标。     

Gli1在鼻咽癌组织中的表达及临床意义

目的:探讨神经胶质瘤相关癌基因1(Gli1)在鼻咽癌组织中的表达及其临床意义.方法:免疫组化方法检测89例鼻咽癌组织及21例正常鼻咽组织中Gli1的表达情况,分析Gli1的表达与与鼻咽癌患者临床病理参数及预后之间的关系.结果:Gli1在鼻咽癌组织的表达水平显著高于正常鼻咽组织(P＜0.01),且Gli1的表达水平与鼻咽癌的T分期、淋巴结转移程度及临床分期显著相关(P＜0.05).此外,Gli1蛋白表达越高,患者的生存时间也明显缩短.结论:Gli1在鼻咽癌组织中呈过表达,可能在鼻咽癌的发生、发展中起重要作用,Gli1有可能成为鼻咽癌治疗的新靶点之一.     

鼻咽癌组织中Gli1的表达及其对5-8F细胞增殖和迁移的影响

目的:研究神经胶质瘤相关癌基因1(Gli1)在鼻咽癌组织中的表达及其对鼻咽癌细胞5-8F增殖和迁移的影响。方法:采用荧光定量PCR和蛋白质印迹法检测40例鼻咽癌组织和12例正常鼻咽组织中Gli1的表达;设计并合成针对Gli1基因的RNAi片段,将其感染5-8F细胞,蛋白质印迹法检测Gli1蛋白表达;MTT实验、平板克隆形成实验、细胞划痕实验和Boyden实验分别研究Gli1基因沉默后对5-8F细胞增殖、迁移和侵袭能力的影响。结果:Gli1在鼻咽癌组织中的表达明显高于正常鼻咽组织,将siRNA-Gli1-01和siRNA-Gli1-02转染入5-8F细胞后,5-8F细胞中Gli1蛋白表达水平明显降低,细胞的增殖、迁移和侵袭能力明显减弱。结论:Gli1基因的表达异常与鼻咽癌的发生发展相关,有望成为鼻咽癌早期诊断和预测预后的生物分子标志物。     

Shh/Gli1信号参与小鼠脊髓损伤后血-脊髓屏障渗透性改变及运动功能恢复

目的:探讨sonic hedgehog/Gli1(Shh/Gli1)信号在小鼠脊髓损伤后病理变化及运动功能恢复中的作用。方法:成年雄性C57野生型、Gli1lz和Gli1lz/lz小鼠行T8节段脊髓夹伤及假手术。Gli1lz小鼠脊髓损伤及假手术后3 d行X-gal染色;7 d行Shh/PDGFr-α、Shh/GFAP、LacZ/GFAP染色,观察Shh/Gli1信号在小鼠脊髓损伤后的激活。C57野生型和Gli1lz/lz小鼠脊髓夹伤后7 d行GFAP染色和实时定量RT-PCR,观察星形胶质细胞反应;术后14 d尾静脉注射伊文氏兰观察血-脊髓屏障改变;术后1、3、5 d和7 d行BMS评分评价小鼠后肢运动功能。结果:脊髓损伤7 d后,损伤区Shh表达升高;Shh/Gli1信号报告基因LacZ表达升高,主要表达于反应性星形胶质细胞;免疫组织化学及实时定量RT-PCR结果显示Gli1基因敲除不影响脊髓损伤后GFAP表达;伊文氏兰染色及其定量分析显示Gli1lz/lz小鼠脊髓损伤后脊髓组织伊文氏兰外渗增加。BMS评分显示Gli1lz/lz小鼠运动功能恢复显著差于野生型小鼠。结论:Shh/Gli1信号在小鼠脊髓损伤后激活,可能参与脊髓损伤后血-脊髓屏障渗透性的改变,并影响脊髓损伤后的运动功能恢复。     

Knockdown of Gli1 by small-interfering RNA enhances the effects of BCNU on the proliferation and apoptosis of glioma U251 cells

Aims: The present study is to investigate the effect of the combination of small-interfering RNA (siRNA) treatment with bis-chloroethylnitrosourea (BCNU) on the proliferation and apoptosis of glioma cells. Methods: According to different treatments, glioma U251 cells were randomly divided into blank group, Lipofectamine group, siRNA-Gli1 group, BCNU group and combination group. After treatments, the morphology of U251 cells was visualized under the microscope. Afterwards, semi-quantitative real-time polymerase chain reaction and Western blotting were used to determine Gli1, Bcl-2, Bax and cyclin D1 mRNA levels and protein expression, respectively. MTT assay was used detect the proliferation of U251 cells, while flow cytometry was performed to determine cell apoptosis and cell cycle. Results: The combination of siRNA-Gli1 and BCNU caused more severe damages to U251 cell shapes compared with siRNA-Gli1 or BCNU alone. The combination of BCNU and siRNA-Gli1 altered mRNA level and protein expression of Bcl-2 and Bax, but not those of Gli1 and cyclin D1. The combination of siRNA-Gli1 and BCNU promoted U251 cell apoptosis. The combination of siRNA-Gli1 and BCNU enhanced the arrestment of U251 cells in G0/G1 phase. The combination of siRNA-Gli1 and BCNU significantly inhibited U251 cell proliferation. Conclusions: The present study demonstrates that combined treatment with siRNA-Gli1 and BCNU significantly inhibits the proliferation and promotes the apoptosis of glioma U251 cells, possibly by the up-regulation of Bax and the down-regulation of Bcl-2. The combination of siRNA-Gli1 and BCNU enhances the inhibition of cell cycles, but does not down-regulate the expression of cell cycle protein cyclin D1.     

Inhibition of SALL4 suppresses carcinogenesis of colorectal cancer via regulating Gli1 expression

Background: SALL4 is a novel oncogene mediating tumorigenesis in multiple carcinomas. However, its actual role and mechanisms participating in the development of colorectal cancer remains unclear. Methods: Immunohistochemical staining and Western blot were conducted to detect the expression of SALL4 and other molecules. siRNA of SALL4 was transfected to silence SALL4 expression in Caco-2 cell line. Flow cytometry was used for cell cycle and apoptosis analysis. Wound healing and transwell assay were used for invasion test. CCK-8 test was employed for cell proliferation and drug sensitivity assessment. Results: By inhibition of SALL4 expression, the proliferation, invasiveness and drug resistance were dramatically reduced while apoptosis rate was up-regulated. Gli1 was found to decrease its expression in SALL4 silencing cells. Moreover, the inhibition on tumorigenesis of Caco-2 by SALL4 silencing was antagonized by Gli1 up-regulation, suggesting Gli1 as a downstream target of SALL4 in cancer development. Conclusion: SALL4 inhibition limited oncogenesis on colorectal cancer by reducing Gli1 expression.