SHH和GL11在早期宫颈癌组织中的表达及意义

目的探讨SHH及神经胶质瘤相关基因同源蛋白1（GL11）在宫颈癌前病变、早期宫颈癌组织中的表达及意义。方法选取早期宫颈癌患者36例、CINHI级28例、正常宫颈20例,使用免疫组化S-P法检测上述组织标本中SHH及GLll的表达。结果SHH蛋白阳性表达率分别为正常宫颈组织20．0％（4／20）、宫颈CINIII60．7％（17／28）、早期宫颈癌69．4％（25／36）,两两比较差异有统计学意义（P均〈0．05）。GL11蛋白阳性表达率分别为正常宫颈组织25．0％（5／20）、宫颈CINⅢ53．6％（15／28）、早期宫颈癌58．3％（21／36）,两两比较差异有统计学意义（P均〈0．05）。结论SHH、GL11过度表达可能参与了宫颈癌的发生发展,其检测有助于宫颈癌的早期诊断。     

SHH信号通路对子痫前期患者滋养细胞凋亡和侵袭的影响及其机制

目的：探讨激活SHH信号通路对子痫前期（PE）患者滋养细胞凋亡和侵袭的影响,并阐明其作用机制。方法：将HTR8/SVneo细胞分为正常对照组、缺氧/复氧（H/R）处理组和purmorphamine+H/R处理组。Western blotting法检测PE胎盘组织和正常妊娠晚期胎盘组织及各组HTR8/SVneo细胞中SHH信号通路相关蛋白SHH、Ptch1、Smo、Gli1和Gli3蛋白表达水平,流式细胞术（FCM）检测各组HTR8/SVneo细胞的凋亡率,Transwell小室法检测各组HTR8/SVneo细胞的穿膜细胞数,Western blotting法检测各组HTR8/SVneo细胞中基质金属蛋白酶2（MMP-2）和基质金属蛋白酶9（MMP-9）蛋白表达水平。结果：PE胎盘组织HTR8/SVneo细胞中SHH、Ptch1、Smo、Gli1和Gli3蛋白表达水平均明显低于正常妊娠晚期胎盘组织（P<0.05）;H/R处理组HTR8/SVneo细胞中SHH、Ptch1、Smo、Gli1、Gli3、MMP-2和MMP-9蛋白表达水平及穿膜细胞数均明显低于正常对照组,细胞凋亡率明显高于正常对照组（P<0.05）;purmorphamine+H/R处理组HTR8/SVneo细胞中SHH、Ptch1、Smo、Gli1、Gli3、MMP-2和MMP-9蛋白表达水平及穿膜细胞数均明显高于H/R处理组（P<0.05）,细胞凋亡率明显低于H/R处理组（P<0.05）。结论：激活SHH信号通路可减少PE患者胎盘组织中HTR8/SVneo细胞凋亡,并通过上调MMP-2和MMP-9表达提高细胞的侵袭能力。     

Expression and significance of sonic hedgehog signaling pathway–related components in brainstem and supratentorial astrocytomas

Background: This study was undertaken to investigate the role of the sonic hedgehog (SHH) signaling pathway in the occurrence of brainstem and supratentorial gliomas by examining the expression of SHH-related cascades in normal brain tissue and brainstem and supratentorial astrocytoma. Methods: Real-time quantitative polymerase chain reaction and immunohistochemistry were used to detect the expression of SHH-related components in 5 normal brain tissue, 10 grade II brainstem glioma, and 10 grade II supratentorial glioma specimens. Results: The mRNA expression levels of SHH-related genes were higher in brainstem astrocytomas than in supratentorial astrocytomas and normal brain tissues. The level of PTCH was statistically significantly higher in brainstem astrocytomas than in supratentorial astrocytomas and normal brain tissues (P < 0.01). Immunohistochemistry semi-quantitative analysis was consistent with the QPCR result that PTCH expression was increased statistically significant in brainstem astrocytomas at the protein level (P <0.05). Conclusion: Enhanced expression of PTCH and activation of the SHH pathway are involved in the occurrence of brainstem glioma. This may be related to the malignant biological behavior difference of brainstem and hemispheric glioma and could be an ideal therapeutic target in cases of brainstem glioma. However, it still cannot be considered a determining factor in the worse prognosis of brainstem glioma than of supratentorial glioma.     

SHH和GLI1在早期宫颈癌组织中的表达及意义

目的 探讨SHH及神经胶质瘤相关基因同源蛋白1(GU1)在宫颈癌前病变、早期宫颈癌组织中的表达及意义.方法 选取早期宫颈癌患者36例、CIN Ⅲ级28例、正常宫颈20例,使用免疫组化S-P法检测上述组织标本中SHH及GLI1的表达.结果 SHH蛋白阳性表达率分别为正常宫颈组织20.0％ (4/20)、宫颈CINⅢ60.7％ (17/28)、早期宫颈癌69.4％ (25/36),两两比较差异有统计学意义(P均＜0.05).GLI1蛋白阳性表达率分别为正常宫颈组织25.0％ (5/20)、宫颈CINⅢ53.6％( 15/28)、早期宫颈癌58.3％(21/36),两两比较差异有统计学意义(P均＜0.05).结论 SHH、GLI1过度表达可能参与了宫颈癌的发生发展,其检测有助于宫颈癌的早期诊断.     

BRD4通过SHH信号通路诱导甲状腺癌细胞增殖、侵袭与迁移

背景与目的：甲状腺癌是目前发病率最高的内分泌系统恶性肿瘤之一,目前的综合治疗手段虽然效果较好,但是部分患者在随后的治疗中会出现继发性摄碘率下降,¹³¹I治疗效果差,从而导致复发及远处转移。近年来的研究发现,溴样结构域蛋白4(double bromodomain-containing protein 4,BRD4)可促进多种恶性肿瘤的进展,因此本研究旨在探究BRD4在甲状腺乳头状癌(papillary thyroid cancer,PTC)细胞中的作用,寻找治疗甲状腺癌的特异性靶点。方法：应用实时荧光定量聚合酶链反应(real-time fluorescent quantitative polymerase chain reaction,RTFQ-PCR)检测PTC组织和癌周组织中BRD4的表达差异,应用siBRD4干扰基因转染PTC细胞株TPC-1,应用蛋白［质］印迹法(Western blot)检验沉默效果,通过MTT实验、平板克隆实验、Transwell小室侵袭与迁移实验检验沉默BRD4前后PTC细胞株TPC-1活力、增殖、迁移及侵袭等生物学行为的变化。进一步应用Western blot及RTFQ-PCR检测钠碘转运体(sodium iodide symporter,NIS)基因及蛋白的表达变化,以及SHH信号通路下游基因SHH、GLI1表达变化情况。结果：BRD4在PTC组织中表达明显增高(P＜0.05);在体外实验中BRD4沉默后PTC细胞株TPC-1的细胞活力、增殖、迁移与侵袭能力下降。此外,BRD4沉默后NIS基因及蛋白表达增高,SHH信号通路下游基因SHH、GLI1表达降低(P＜0.05)。结论：BRD4通过上调SHH信号通路相关基因促进PTC细胞的侵袭与迁移,沉默BRD4可以促进NIS的表达,BRD4有望成为治疗甲状腺癌的新靶点。     

Assessment of the stromal contribution to Sonic Hedgehog-dependent pancreatic adenocarcinoma

Pancreatic ductal adenocarcinoma (PDAC) remains one of the most lethal malignancies. It is typically detected at an advanced stage, at which the therapeutic options are very limited. One remarkable feature of PDAC that contributes to its resilience to treatment is the extreme stromal activation seen in these tumors. Often, the vast majority of tumor bulk consists of non-tumor cells that together provide a tumor-promoting environment. One of the signals that maintains and activates the stroma is the developmental protein Sonic Hedgehog (SHH). As the disease progresses, tumor cells produce increasing amounts of SHH, which activates the surrounding stroma to aid in tumor progression. To better understand this response and identify targets for inhibition, we aimed to elucidate the proteins that mediate the SHH-driven stromal response in PDAC. For this a novel mixed-species coculture model was set up in which the cancer cells are human, and the stroma is modeled by mouse fibroblasts. In conjunction with next-generation sequencing we were able to use the sequence difference between these species to genetically distinguish between the epithelial and stromal responses to SHH. The stromal SHH-dependent genes from this analysis were validated and their relevance for human disease was subsequently determined in two independent patient cohorts. In non-microdissected tissue from PDAC patients, in which a large amount of stroma is present, the targets were confirmed to associate with tumor stroma versus normal pancreatic tissue. Patient survival analysis and immunohistochemistry identified CDA, EDIL3, ITGB4, PLAUR and SPOCK1 as SHH-dependent stromal factors that are associated with poor prognosis in PDAC patients. Summarizing, the presented data provide insight into the role of the activated stroma in PDAC, and how SHH acts to mediate this response. In addition, the study has yielded several candidates that are interesting therapeutic targets for a disease for which treatment options are still inadequate.     

牙源性角化囊肿SMO基因突变检测

目的检测牙源性角化囊肿(odontogenic keratocyst,OKC)是否存在SMO基因突变,进一步完善对OKC发病机制的认识。方法收集2012年9月至2017年6月就诊于北京大学口腔医学院·口腔医院口腔颌面外科的OKC患者,10例为痣样基底细胞癌综合征性OKC(女性4例,男性6例),20例为散发性OKC(女性7例,男性13例)。采集患者的病变组织,分离衬里上皮和纤维间质,采用Sanger测序法分别检测上皮与间质DNA中SMO基因突变情况。结果检测发现3个SMO基因突变位点,即1例综合征性OKC携带c.2081C>G(p.P694R)突变,2例散发性OKC分别携带c.907C>T(p.L303F)突变和c.1247_1248delinsAA(p.G416E)突变,前2例突变为未被报道过的SMO新突变,且2例散发性OKC均不伴PTCH1突变。结论除PTCH1突变外,OKC还存在SMO基因突变,可能与OKC的发病机制有关。     

The stromal compartments in pancreatic cancer: Are there any therapeutic targets?

Pancreatic ductal adenocarcinoma (PDAC) is characterised by an abundant stromal response also known as a desmoplastic reaction. Pancreatic Stellate Cells have been identified as playing a key role in pancreatic cancer desmoplasia. There is accumulating evidence that the stroma contributes to tumour progression and to the low therapeutic response of PDAC patients. In this review we described the main actors of the desmoplastic reaction within PDAC and novel therapeutic approaches that are being tested to block the detrimental function of the stroma.