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Hh信号通路相关因子Shh、Ptch1、Gli1 mRNA的表达及意义
编辑人员丨4天前
目的:探讨前列腺癌、癌旁前列腺组织以及正常前列腺增生组织中Hedgehog(Hh)信号通路相关因子Shh、Ptch1、Gli1 mRNA的表达及意义。方法:选择2017年2月至2019年2月本院收治的42例前列腺癌患者作为研究对象,设为观察组。所有患者均拟手术治疗,术中取癌组织与癌旁组织(距离病灶≥3 cm);选择同期手术治疗的39例前列腺增生患者的手术标本,设为对照组。利用免疫组化和qRT-PCR技术检测前列腺增生、前列腺癌和癌旁组织中Hh信号通路相关因子Shh、Ptch1和Gli1蛋白及mRNA的表达情况,分析比较Hh信号通路在前列腺增生、前列腺癌和癌旁组织中表达的差异及其机制。结果:观察组的癌旁组织与对照组的Shh、Ptch1和Gli1蛋白阳性率比较,差异均无统计学意义(均 P>0.05);观察组的癌组织中Shh、Ptch1和Gli1蛋白阳性率均高于对照组(均 P<0.05)。观察组的癌旁组织与对照组的Shh、Ptch1和Gli1 mRNA表达水平比较,差异均无统计学意义(均 P>0.05);观察组的癌组织中Shh、Ptch1和Gli1 mRNA表达水平均高于观察组的癌旁组织和对照组(均 P<0.05);Spearman秩相关系数结果显示,Hh信号通路中Shh、Ptch1和Gli1与术前前列腺特异抗原(PSA)均无相关性(均 P>0.05);与Gleason评分、临床分期、病理分级均呈正相关性(均 P<0.05)。 结论:Hh信号通路中Shh、Ptch1和Gli1在前列腺癌患者中呈高表达,其在前列腺癌疾病的发生发展中可能起到一定的作用。
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编辑人员丨4天前
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孕期邻苯二甲酸二丁酯暴露抑制Hedgehog信号导致后代肾脏自噬
编辑人员丨4天前
目的:观察孕期邻苯二甲酸二丁酯(DBP)暴露对后代肾脏的毒理作用,并探讨其机制。方法:将6只孕鼠在孕期14~18 d分别随机予以850 mg/(kg·d)的DBP处理3只孕鼠为染毒组,另外3只孕鼠予以等剂量的玉米油处理为对照组。在产后第1天分别随机收集各组6只子代鼠的肾脏组织,采用免疫组织化学(IHC)染色检测肾脏自噬水平,蛋白质印迹法(Western blot)及定量分析验证自噬相关基因Beclin-1在肾脏中的表达。用Western blot及实时定量反转录聚合酶链反应(RT-qPCR)检测肾脏组织中Hedgehog信号通路上的相关标志物的表达水平。在体外实验中,分别予以Hedgehog信号通路的抑制剂索尼德吉Sonidegib及激动剂重组音刺猬蛋白(SHH)处理NRK52E肾小管上皮细胞,采用Western blot分析各组的自噬水平。组间比较采用独立 t检验。 结果:IHC提示染毒组Beclin-1的表达高于对照组(2.37±0.12比1.00±0.10, t=15.190, P<0.05),蛋白印记及其定量分析显示染毒组Beclin-1蛋白表达量高于对照组(1.20±0.104比1.00±0.01, t=3.317, P<0.01)。同时,Western blot及其定量分析表明,暴露组Hedgehog信号通路标记物Gli1和Ptch1表达低于对照组(1.00±0.15比0.43±0.09, t=5.644, P<0.01;1.00±0.01比0.35±0.10, t=11.20, P<0.01)。RT-qPCR也提示暴露组Gli1和Ptch1表达低于对照组(1.00±0.18比0.51±0.10, t=4.122, P<0.01;1.00±0.10比0.62±0.08, t=5.140, P<0.01)。体外研究证实当加入Hedgehog抑制剂Sonidegib后,实验组的NRK52E细胞自噬指数(LC3Ⅱ/LC3Ⅰ)高于对照组(0.49±0.11比1.25±0.05, t=10.890, P<0.01),而当加入其激动剂SHH后,实验组的自噬指数低于对照组(0.49±0.11比0.36±0.09, t=11.260, P<0.01)。 结论:孕期DBP暴露通过抑制Hedgehog信号引起后代肾脏发生自噬,从而可能引起肾脏相关疾病。
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编辑人员丨4天前
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Genetic Predisposition to Numerous Large Ulcerating Basal Cell Carcinomas and Response to Immune Therapy
编辑人员丨4天前
Objective::Well-defined germ-line mutations in the PTCH1 gene are associated with syndromic multiple basal cell carcinomas (BCCs). Here, we used whole exome sequencing (WES) to identify the role of patched-1 in patients with multiple, unusually large BCCs. Methods::A 72-year old patient presenting with numerous BCCs progressing to large ulcerating lesions was enrolled. WES was used to identify the pathogenic gene locus.Results::Genetic work-up by WES identified a homozygous PTCH1 nonsense mutation in the tumor tissue but not present in her blood cells or in non-lesional skin. In addition, heterozygous missense mutations were identified in three cancer-associated genes ( EPHB2, RET, and GALNT12) in blood cells as well as in lesional and non-lesional skin. We also tested systemic immune therapy as a potentially beneficial approach to treat patients with numerous large BCCs on scatted areas of involvement. A rapid and sustained response to nivolumab was noted, suggesting that it is an efficacious drug for long-term therapeutic outcome. Conclusion::PTCH1, EPHB2, RET, and GALNT12 may potentially contribute to the synergistic oncogene driven malignant transformation manifesting as multiple, unusually large BCCs.
immune therapy malignant transformation gene-susceptibility non-syndromic basal cell carcinoma PTCH1...不再出现此类内容
编辑人员丨4天前
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Genetic insights into thymic carcinomas and thymic neuroendocrine neoplasms denote prognosis signatures and pathways
编辑人员丨4天前
Background::Thymic carcinomas (TCs) and thymic neuroendocrine neoplasms (TNENs) are two aggressive subtypes of thymic malignancy. Traditional therapy for advanced TCs and TNENs has limited outcome. New genomic profiling of TCs and TNENs might provide insights that contribute to the development of new treatment approaches.Methods::We used gene panel sequencing technologies to investigate the genetic aberrations of 32 TC patients and 15 TNEN patients who underwent surgery at Shanghai Chest Hospital between 2015 and 2017. Patient samples were sequenced using a 324-gene platform with licensed technologies. In this study, we focused on clinically relevant genomic alterations (CRGAs), which are previously proven to be pathogenic alterations, to identify the pathology-specific mutational patterns, prognostic signatures of TCs and TNENs.Results::The mutational profiles between TCs and TNENs were diverse. The genetic alterations that ranked highest in TCs were in CDKN2A, TP53, ASXL1, CDKN2B, PIK3C2G, PTCH1, and ROS1, while those in TNENs were in MEN1, MLL2, APC, RB1, and TSC2. Prognostic analysis showed that mutations of ROS1, CDKN2A, CDKN2B, BRAF, and BAP1 were significantly associated with worse outcomes in TC patients, and that mutation of ERBB2 indicated shortened disease-free survival (DFS) and overall survival (OS) in TNEN patients. Further investigation found that the prognosis-related genes were focused on signal pathways of cell cycle control, chromatin remodeling/DNA methylation, phosphoinositide 3-kinases (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR), and receptor tyrosine kinase (RTK)/RAS/mitogen-activated protein kinase (MAPK) signaling. Conclusion::We profiled the mutational features of 47 Chinese patients with thymic malignancy of diverse pathologic phenotypes to uncover the integrated genomic landscape of these rare tumors, and identified the pathology-specific mutational patterns, prognostic signatures, and potential therapeutic targets for TCs and TNENs.
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编辑人员丨4天前
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牙源性角化囊肿SMO基因突变检测
编辑人员丨4天前
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。
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编辑人员丨4天前
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SETD2在恶性胸膜间皮瘤中的表达与临床意义研究
编辑人员丨4天前
目的:分析恶性胸膜间皮瘤(MPM)基因突变概况和高频突变基因的表达水平,筛选出影响MPM患者预后的关键分子和临床病理因素。方法:于2020年3月,分别从癌症基因组图谱(TCGA)数据库和人类肿瘤相关基因表达汇编(GEO)数据库下载86例MPM组织的基因二代测序数据和89例MPM组织样本的基因芯片表达数据,收集相关体细胞突变信息及对应的临床病理资料。汇总TCGA数据库中组织样本基因突变概况并分析高频突变基因表达水平与MPM患者临床病理学特征、石棉接触史和预后的关系,筛选出与MPM预后显著相关的基因进行基因集富集分析(GSEA)。利用GEO数据库的芯片表达数据对筛选出的关键基因和临床病理特征进行生存分析和GSEA验证。结果:TCGA数据集中单核苷酸变异(SNV)频率最高的10个基因是BAP1、NF2、TP53、TTN、SETD2、LATS2、CCDC168、FAT4、PTCH1和ZNF469。野生型NF2、TP53、SETD2和CCDC168基因的高表达率高于突变型,差异均具有统计学意义( P<0.05)。TCGA数据集Cox多因素分析结果显示,上皮型( HR=0.425,95% CI:0.235~0.767, P<0.01)和SETD2低表达( HR=0.516,95% CI:0.307~0.868, P=0.011)MPM患者的死亡风险较低。GEO数据集生存分析验证发现,上皮型MPM患者生存时间更长,肉瘤型MPM患者生存时间最短( P<0.01)。TCGA和GEO数据集富集结果显示STED2可能与G2M细胞周期检查点、E2F靶标基因、MYC靶标基因、蛋白分泌、有丝分裂纺锤体、MTORC1通路、TGF-β通路、雄激素反应和紫外线反应等通路相关。 结论:MPM伴随着以BAP1、NF2、TP53、TTN、SETD2和LATS2等基因为代表的较高频率的基因突变。SETD2表达水平和组织类型中的上皮型可能是影响MPM预后的因素。
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编辑人员丨4天前
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胶质瘤相关癌基因1特异性阻断剂对结直肠癌肿瘤干细胞分化的影响及对Hedgehog信号通路的调控作用
编辑人员丨4天前
目的:观察胶质瘤相关癌基因1特异性阻断剂(GANT-61)对结直肠癌肿瘤干细胞(CRC-CSCs)分化及对Hedgehog信号通路的调控作用的影响。方法:磁珠分选法获得CD133 + CRC-CSCs,用含不同浓度(0、10、20、40 μmol/L)的GANT-61的干细胞培养液干预,设为对照组、低、中、高剂量组,测定干预不同时刻细胞活性、CRC-CSCs球形成率、CD133 +占比、核转录因子1(Gli1)、Hh受体蛋白(Ptch1)、c-Myc、细胞角蛋白20(CK20) mRNA和蛋白表达。多样本计量资料比较采用单因素方差分析,两两样本比较采用SNK- q检验。 结果:与对照组比较,GANT-61各剂量组不同时刻CCK-8试验吸光度( A)值均降低(24 h: t=2.445、4.985、6.903;48 h: t=2.859、5.270、7.488;72 h: t=3.324、8.053、12.042, P<0.05),且随GANT-61干预剂量升高而降低( F=13.382、16.842、41.053, P<0.05),差异有统计学意义;对照组、低、中剂量组CCK-8试验 A值随干预时间延长升高,且干预不同时刻CCK-8试验 A值比较差异均有统计学意义( F=44.598、31.844, P<0.05),而高剂量组随干预时间延长无明显变化( F=1.585, P>0.05),差异无统计学意义。对照组、GANT-61低、中、高剂量组CRC-CSCs球形成率分别为(13.57±1.55)%、(11.22±1.01)%、(5.46±0.67)%、(1.60±0.30)%,CD133 +占比分别为(96.20±3.07)%、(81.34±5.84)%、(62.01±5.03)%、(45.38±4.21)%;与对照组比较,GANT-61各剂量组CRC-CSCs球形成率及CD133 +占比均降低(CRC-CSCs球形成率: t=2.840、10.739、16.954, P<0.05;CD133 +占比: t=5.036、12.974、21.809, P<0.05),且随GANT-61干预剂量升高而降低,各剂量组组间比较差异均有统计学意义(CRC-CSCs球形成率: F=225.500, P<0.05;CD133 +占比: F=62.988, P<0.05)。与对照组比较,GANT-61各剂量组Gli1、Ptch1、c-Myc、CK20 mRNA和蛋白相对表达量均降低(mRNA:低剂量组 t=2.577、2.659、2.524、4.031;中剂量组 t=5.271、5.587、6.163、6.500;高剂量组 t=7.442、8.976、8.750、9.632, P<0.05;蛋白:低剂量组 t=3.008、2.517、2.385、2.981;中剂量组 t=5.664、8.923、9.500、11.180;高剂量组 t=14.435、14.137、15.727、14.142, P<0.05),且随GANT-61干预剂量升高而降低(mRNA: F=14.825、21.057、22.389、27.160, P<0.05;蛋白: F=65.340、95.789、151.230、119.877, P<0.05),差异有统计学意义。 结论:GANT-61可能通过下调Hedgehog信号通路基因Gli1、Ptch1、靶基因c-Myc、分化基因CK20 mRNA和蛋白的表达,从而起到抑制CRC-CSCs细胞自我更新能力、促进细胞分化作用。
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编辑人员丨4天前
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Association between PTCH1 gene polymorphisms and chronic obstructive pulmonary disease susceptibility in a Chinese Han population: a case-control study
编辑人员丨4天前
Background::Chronic obstructive pulmonary disease (COPD) is a leading cause of morbidity and mortality worldwide. Genome-wide association studies in non-Asian population revealed a link between COPD and mutations in the PTCH1 gene encoding Patched1, a receptor in the Hedgehog signaling pathway important for lung morphogenesis and pulmonary function. The aim of this study was to investigate the association between PTCH1 polymorphisms and the COPD risk in the Chinese Han population. Methods::We performed a case-control study including 296 patients with COPD and 300 healthy individuals. Single-nucleotide polymorphisms in the PTCH1 gene were identified and genotyped based on the linkage disequilibrium analysis in all participants. Odds ratios (ORs) and 95% confidence intervals (95% CIs) were estimated using logistic regression analysis after adjustment for age, gender, and smoking. Results::In total, 28 single-nucleotide polymorphisms were identified in patients with COPD. Among them, "A" allele of rs28491365 (OR: 1.388, 95% CI: 1.055-1.827, P = 0.018), and "G" alleles of rs10512248 (OR: 1.299, 95% CI: 1.021-1.653, P = 0.033) and rs28705285 (OR: 1.359, 95% CI: 1.024-1.803, P = 0.033; respectively) were significantly associated with an increased COPD risk. Genetic model analysis revealed that the "T/T" genotype of rs34695652 was associated with a decreased COPD risk under the recessive model (OR: 0.490, 95% CI: 0.270-0.880, P = 0.010), whereas rs28504650/rs10512248 haplotype CG was significantly associated with an increased COPD risk after adjustment for age, gender, and smoking status (OR: 6.364, 95% CI: 1.220-33.292, P = 0.028). Conclusions::The study provides a new insight into the role of PTCH1 polymorphisms in the susceptibility to COPD in the Chinese Han population.
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编辑人员丨4天前
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髓母细胞瘤相关遗传综合征的研究进展
编辑人员丨4天前
髓母细胞瘤是儿童期颅内高发肿瘤之一,分为Wnt亚型、Shh亚型、Group 3和Group 4亚型。手术切除肿瘤后可以辅以放疗和化疗,患者的5年无进展生存率可达50%~70%以上。部分患者伴有一些遗传性疾病,例如Gorlin-Goltz综合征、Li-Fraumeni综合征、家族性腺瘤性息肉病综合征等,不同综合征伴随着不同的胚系基因突变,其治疗方案、临床预后与患者的年龄、病理、分子亚型有密切关系。Shh亚型是合并基因胚系突变的高危亚型,需要常规行基因检测,以排除 TP53、 PTCH1、 SUFU、 GPR161、 ELP1等基因胚系突变,其他亚型的发病率相对较低,但需要排除 APC、 PALB2、 BRCA2等基因胚系突变。
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编辑人员丨4天前
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错配修复缺陷与完整的结直肠癌相关基因体细胞突变差异分析
编辑人员丨4天前
目的:分析DNA错配修复(MMR)蛋白缺陷(dMMR)和完整(pMMR)的结直肠癌(CRC)患者相关基因体细胞突变差异。方法:收集2015年1月至2017年1月于浙江大学医学院附属第二医院行手术治疗、术后常规免疫组化检测报告为dMMR的93例CRC患者石蜡病理组织行二代测序技术检测。采用免疫组化法检测93例CRC患者肿瘤组织中4种MMR蛋白(MLH1、MSH2、MSH6和PMS2)的表达情况,依据美国病理学家协会(CAP)标准对免疫组化检测结果进行重新判读。采用二代测序技术检测93例CRC患者石蜡病理组织中41个基因的体细胞突变情况,采用Fisher精确检验分析组间基因突变差异。结果:依据CAP标准重新判读后,93例CRC患者中pMMR组31例,dMMR组62例。dMMR组基因突变中位数为9.5个,高于pMMR组(3.0个, P<0.001)。dMMR组和pMMR组中17个基因存在体细胞突变差异,分别为乳腺癌易感基因1(BRCA1)、BRCA2、MLH1、PDGFRA、PIK3CA、APC、ATM、KIT、MET、PMS2、MSH6、POLE、MSH2、PTCH1、表皮生长因子受体(EGFR)、TP53和ERBB2基因。dMMR组致病体细胞BRAF、MLH1、MSH2和MSH6基因突变率高于pMMR组[分别为21.0%(13/62)和9.7%(3/31),9.7%(6/62)和0(0/31),21.0%(13/62)和0(0/31),22.6%(14/62)和0(0/31),均 P<0.05]。BLM N515fs、BRAF V600E、PTCH1 R1308fs和KRAS G13D位点突变率在dMMR组和pMMR组CRC中差异均有统计学意义[分别为22.6%(14/62)和0(0/31),19.4%(12/62)和3.2%(1/31),11.3%(7/62)和0(0/31),16.1%(10/62)和3.2%(1/31),均 P<0.05]。在MLH1和PMS2共同缺失的dMMR患者中3个不常见位点BLM N515fs、MSH6 F1088fs和PTCH1 R1308fs的突变率分别为28.2%(11/39)、15.4%(6/39)和15.4%(6/39),与pMMR患者[均为0(0/31)]比较,差异均有统计学意义(均 P<0.05)。 结论:dMMR结直肠癌患者有更多的相关基因发生体细胞突变。BRAF V600E突变与dMMR密切相关。KRAS G13D、BLM N515fs和PTCH1 R1308fs突变位点也与MMR蛋白的表达有关。
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编辑人员丨4天前
