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。     

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。     

Gli1抑制剂GANT61对小鼠牙胚体外发育的影响

目的:研究外源性Gli1抑制剂GANT61对小鼠磨牙胚体外发育的影响。方法:采用小鼠磨牙胚体外培养系统,运用体视镜、组织学HE染色方式观察小鼠磨牙胚被Gli1抑制剂GANT61影响后的发育情况,并对牙胚构成部分进行测量。结果:在Gli1抑制剂GANT61作用下,实验组相对于对照组出现下磨牙牙胚发育速度减慢,成釉器凹陷且形态不规则,内釉上皮细胞呈现不规则排列,成釉器大小在高浓度GANT61组明显变小。结论:Gli1抑制剂GANT61可能通过扰乱成釉器的发育阻碍胎鼠磨牙胚的发育过程。     

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.     

肝细胞肝癌中Hedgehog信号通路效应蛋白的功能分析

目的 探讨Gli1、Gli3蛋白在肝细胞癌组织中的表达及临床意义.方法 免疫组化法检测36例肝细胞癌组织及其癌旁肝组织中Gli1、Gli3蛋白的表达,并与临床因素进行相关性分析.结果 在肝细胞癌组织及癌旁肝组织中Gli1的阳性表达率分别为75％和36.1％,Gli3的阳性表达率分别为58.3％和30.6％.Gli1和Gli3在肝细胞癌组织中的阳性表达率显著高于癌旁肝组织(P＜0.05).Gli1和Gli3两种蛋白的表达呈正相关(r=0.423,P＜0.05).Gli3的表达与临床指标(年龄、肿瘤大小、分化和淋巴结转移)无关.Gli1的表达与年龄、肿瘤大小无关,但与肿瘤分化程度和淋巴结转移相关(P＜0.05).结论 Gli1、Gli3蛋白在肝细胞癌组织中有较高的阳性表达率且具有相关性,且Gli1的表达率与肿瘤分化程度和淋巴结转移相关.检测Gli1、Gli3蛋白表达有助于肝细胞癌的诊断,可能作为判断肝细胞癌恶性程度及预后的指标.     

Shh/Gli1信号通路在异氟醚后处理减轻大鼠脑缺血-再灌注损伤中的作用

目的评价Shh/Gli1信号通路在异氟醚后处理减轻大鼠脑缺血-再灌注损伤中的作用。方法清洁级健康雄性SD大鼠44只,6~8周龄,体重220~280 g,采用随机数字表法将其分为四组:假手术组(S组)、缺血-再灌注组(IR组)、缺血-再灌注+异氟醚后处理组(ISO组)和环巴胺+缺血-再灌注+异氟醚后处理组(CYC组),每组11只。采用线栓法栓塞大脑中动脉90 min、再灌注24 h制备脑缺血-再灌注损伤模型。ISO组大鼠在再灌注即刻吸入1.5%异氟醚。CYC组大鼠在缺血前30 min腹腔注射Shh/Gli1信号通路特异性抑制剂环巴胺10 mg/kg。再灌注24 h后,所有大鼠进行神经行为学评分,采用TTC法测定脑梗死体积,HE染色和尼氏染色观察病理学改变,TUNEL染色观察海马CA1区细胞凋亡,免疫荧光和Western blot法测定Shh和Gli1蛋白含量。结果与S组比较,IR组、ISO组和CYC组大鼠神经行为学评分明显升高,脑梗死体积明显增大,坏死和凋亡细胞明显增多,组织病理学损伤严重,Shh和Gli1蛋白含量明显增加(P<0.05)。与IR组比较,ISO组神经行为学评分和脑梗死体积明显降低,坏死和凋亡明显减少,组织病理学损伤明显减轻,Shh和Gli1蛋白含量明显增加(P<0.05)。与ISO组比较,CYC组神经行为学评分明显升高,脑梗死体积明显增大,坏死和凋亡细胞明显增多,组织病理学损伤明显加重,Shh和Gli1蛋白含量明显减少(P<0.05)。结论 Shh/Gli1信号通路激活参与了异氟醚后处理减轻大鼠脑缺血-再灌注损伤的过程。