S100A8基因在喉鳞癌发生中作用机制的研究

目的：研究S100A8与喉癌发生的关系,揭示炎症在喉癌发生中的作用.方法：应用RT-PCR、West-ern Blot、免疫组化检测S100A8表达,RNAi方法检测其功能、Transwell检测细胞侵袭力,TUNEL和流式细胞仪检测细胞凋亡,MTT检测细胞生长活性.结果：36例喉鳞癌组织中17例出现S100A8 mRNA表达上调（47.2%）,而7例喉部良性肿瘤中2例出现表达上调（28.5%）.13例病人喉鳞癌组织中7例S100A8蛋白表达增高.免疫组化分析喉鳞癌组织S100A8蛋白阳性率为54.3%.在高分化喉鳞癌组织为71.4%,而分化程度差的喉癌组织中阳性率为12.8%（P=0.023）.RNAi抑制S100A8基因使Hep2细胞凋亡率增高近9倍,使Hep2细胞侵袭力下降,影响Bel-2基因表达,但对Hep2细胞的IKKα表达无明显影响.结论：S100A8与喉鳞癌发生相关并参与喉鳞癌分化,S100A8基因具有抑制喉癌细胞凋亡,促进Hep2细胞侵袭的作用.S100A8基因可能部分通过NF-κB通路参与喉癌发生、发展,在此通路中S100A8基因位于p65下游.     

RNAi抑制NF—κB通路相关基因对喉鳞癌凋亡及转移的影响

目的：用RNA干涉方法探讨NF—κB通路参与喉癌发生、发展的可能机制。方法：RT—PCR、West—ern Blot检测喉鳞癌组织中NF—κB通路中相关基因表达，RNA干涉方法抑制p65，p50表达，流式细胞仪，TUNEL方法检测细胞凋亡，Transwell检测细胞侵袭力。结果：36例喉鳞癌组织中21例p65 mRNA表达上调（58．33％）高于癌旁组织（P=0，012），13例出现p50 mRNA表达上调（36.11％），但与癌旁组织比较无显著差异（P=0．602）。喉鳞癌组织的p65蛋白表达明显较癌旁组织增强（P=0．044），而p50蛋白表达无显著差异。特异siRNA转染Hep2细胞明显抑制p50，p65基因表达，该作用可持续10天。RNAi抑制p65基因表达使Hep2细胞凋亡率增加，在转染后第7天最高达29．89％，与对照组比较升高超过10倍（P=0．020）；同时使Hep2细胞侵袭力明显下降（P=0．003），而干涉p50基因对细胞凋亡及细胞侵袭性影响不明显。结论：RNAi抑制p65表达诱导Hep2细胞凋亡、抑制细胞侵袭力，提示抑制p65表达与常规治疗结合可能成为改善喉癌预后的选择。     

S100A4在食管鳞癌组织中的表达及意义

目的:研究S100A4在食管鳞癌组织中的表达及意义,探讨它在食管鳞癌的侵袭和转移中的作用。方法:采用SP免疫组织化学方法,检测53例食管癌患者手术切除标本中癌灶及34例癌远旁食管组织的S100A4蛋白的表达,利用统计软件SPSS13.0进行统计学分析,探讨其在性别、年龄、肿瘤大小、浸润程度、淋巴结转移、分化程度及临床分期中的关系。结果:S100A4在食管鳞癌中表达显著高于癌远旁正常食管组织(P<0.05)。S100A4表达与食管鳞癌临床病理特征关系密切,在有淋巴结转移组与无转移组中有显著性差异(P<0.05)。在食管癌不同临床TNM分期中,S100A4阳性表达随临床分期的升高而明显增加,各期间均有显著性差异(P<0.05)。S100A4表达与食管鳞癌分化程度相关,随分化程度的降低,S100A4阳性率升高,但只有在高、中分化组与低分化组进行比较有显著差异(P<0.05)。S100A4的表达与肿瘤浸润有密切的正相关关系,随肿瘤浸润程度的增加,S100A4的阳性率升高(P<0.05)。S100A4在食管鳞癌中表达与患者性别、年龄及肿瘤大小无关。结论:S100A4与食管鳞癌的侵袭和转移有关;S100A4的表达与淋巴结转移密切相关,有可能为食管癌的靶向治疗开辟新的途径。     

S100A8和S100A9在子宫颈鳞癌组织中的表达

目的:探讨S100A8、S100A9在子宫颈鳞癌及正常宫颈组织中的表达及其意义。方法:采用半定量的RT-PCR技术和免疫组织化学的方法,从mRNA及蛋白水平检测32例子宫颈鳞癌和15例正常宫颈组织中S100A8、S100A9的表达,及与不同临床病理参数的关系。结果:子宫颈鳞癌组织中,S100A8、S100A9在mRNA及蛋白水平上均低表达(P<0.05),其中,低分化肿瘤的表达明显低于高、中分化肿瘤(P<0.05)。结论:S100A8、S100A9在子宫颈鳞癌组织中低表达,且可能与鳞状细胞的分化程度相关。提示S100A8、S100A9低表达可能与子宫颈鳞癌的发生发展有关系。     

miRNA-24下调S100A8蛋白表达能够抑制人喉鳞癌Hep2细胞侵袭

目的：探讨microRNA-24（miRNA-24）对喉鳞状细胞癌（laryngeal squamous carcinoma,LSCC）Hep2细胞侵袭影响的分子机制。方法：应用miRanda和RNA22靶基因预测软件预测miR-24的靶基因及其结合位点,根据预测结果通过转染miR-24前体上调其在Hep2细胞中的表达,应用荧光定量PCR和Western blot分别检测miR-24过表达对S100钙结合蛋白A8（S100 calcium binding protein A8,S100A8）mRNA和蛋白表达变化的影响。S100A8抗体阻断方法,检测转染miR-24前体后Hep2细胞侵袭能力的变化。结果：S100A83＇非翻译区（3＇-untranslation region,3＇UTR）含有一个miR-24结合位点,而且该位点与大鼠的同源性高达95%;miR-24基因转染的Hep2细胞中miR-24的表达显著升高,S100 A8蛋白的表达显著降低,（P均〈0.05）,而S100A8 mRNA表达变化未见显著性差异,（P〉0.05）。与未阻断组相比,miR-24前体转染能明显降低阻断S100A8组的Hep2细胞侵袭能力,（P〈0.05）。结论：LSCC Hep2细胞中,miR-24可结合到S100A8基因的3＇UTR,在转录后水平上负性调控S100A8基因的表达,从而抑制Hep2细胞侵袭。     

miRNA-24下调S100A8蛋白表达能够抑制人喉鳞癌Hep2细胞侵袭

目的:探讨microRNA-24(miRNA-24)对喉鳞状细胞癌(laryngeal squamous carcinoma,LSCC)Hep2细胞侵袭影响的分子机制。方法:应用miRanda和RNA22靶基因预测软件预测miR-24的靶基因及其结合位点,根据预测结果通过转染miR-24前体上调其在Hep2细胞中的表达,应用荧光定量PCR和Western blot分别检测miR-24过表达对S100钙结合蛋白A8(S100 calcium binding protein A8,S100A8)mRNA和蛋白表达变化的影响。S100A8抗体阻断方法,检测转染miR-24前体后Hep2细胞侵袭能力的变化。结果:S100A83’非翻译区(3’-untranslation region,3’UTR)含有一个miR-24结合位点,而且该位点与大鼠的同源性高达95%;miR-24基因转染的Hep2细胞中miR-24的表达显著升高,S100 A8蛋白的表达显著降低,(P均<0.05),而S100A8 mRNA表达变化未见显著性差异,(P>0.05)。与未阻断组相比,miR-24前体转染能明显降低阻断S100A8组的Hep2细胞侵袭能力,(P<0.05)。结论:LSCC Hep2细胞中,miR-24可结合到S100A8基因的3’UTR,在转录后水平上负性调控S100A8基因的表达,从而抑制Hep2细胞侵袭。     

胰腺癌患者血清中S100A8与S100A9水平及其临床意义

目的检测胰腺癌患者血清S100A8与S100A9的水平及探讨其临床意义。方法收集胰腺癌患者90例,同期纳入体检健康者50例为对照组。检测两组研究者血清中S100A8和S100A9的水平。分析胰腺癌患者血清S100A8和S100A9水平与其临床病理特征的相关性。对所有患者疗后随访6个月,比较死亡组与存活组患者血清中S100A8和S100A9水平。结果胰腺癌患者血清中S100A8和S100A9水平显著高于对照组(P<0.01)。胰腺癌患者血清中S100A8和S100A9水平与患者年龄、性别、肿瘤部位无显著相关(P>0.05),与其分化程度、临床分期呈明显相关(P<0.01)。6个月随访结果:27例死亡,18例存活,其中死亡组患者入院时血清S100A8和S100A9水平明显高于存活组,比较有显著性差异(P<0.01)。结论胰腺癌患者血清中S100A8和S100A9水平明显提高,且与其恶性程度密切相关,可能对临床评估胰腺癌预后有一定参考价值。     

S100A16在食管鳞癌中的表达及其对肿瘤细胞迁移和侵袭能力的影响

目的 探讨S100A16在食管鳞状细胞癌(ESCC)组织中的表达及其对ESCC细胞迁移、侵袭能力的影响。方法 利用生物信息学分析S100A16在ESCC中的表达趋势及生存情况,利用免疫组化进一步验证S100A16在ESCC患者癌组织中的表达,构建S100A16-overexpression和S100A16-siRNA细胞模型,采用细胞划痕实验及细胞侵袭实验观察S100A16对TE12细胞迁移能力及侵袭能力的影响。结果 GEO数据库中分析ESCC GSE26886芯片,发现S100A16在ESCC中表达上调;而在TCGA和GTEx数据库中分析发现,S100A16在ESCC中表达下调。利用基因表达谱动态分析(GEPIA)在线分析网站进行生存分析,结果显示S100A16对食管癌患者的总生存期及无病生存期均无显著影响(P>0.05)。S100A16在ESCC癌组织中的低表达率和高表达率分别为65.0%(52/80)、35.0%(28/80),S100A16在癌旁组织中的低表达率和高表达率分别为40.0%(32/80)、60.0%(48/80);S100A16在ESCC癌组织中低表达,与癌...     

S100A4与MMP-9在喉鳞癌中的表达和相关性研究

分析正常喉组织、喉鳞癌组织中S100A4和MMP-9的表达及临床意义,探讨其在喉鳞癌发生发展中的作用及相关性。方法：采用免疫组织化学SP法检测S100A4和MMP-9蛋白的表达并分析二者的相关性及其与临床病理因素的关系;采用Western blot法检测S100A4和MMP-9蛋白的表达。结果：免疫组化结果表明S100A4和MMP-9蛋白在癌旁喉组织和喉鳞癌中的阳性表达率均呈升高趋势,分别为8.6%和14.3%,63.8%和71.3%,并且二者表达均与喉鳞癌淋巴结转移相关。统计学结果显示80例喉鳞癌组织中,S100A4和MMP-9蛋白表达呈正相关（P<0.01,r=0.440）。Western blot结果表明S100A4和MMP-9蛋白在喉鳞癌组织中表达高于癌旁喉组织。在喉癌细胞系Hep-2中干扰S100A4蛋白的表达后,MMP-9的蛋白水平下调。结论：S100A4和MMP-9在喉鳞癌发生发展中均存在异常,并且S100A4基因抑制后能够显著下调MMP-9的表达。     

血清S100A8和S100A9对原发性肝癌的临床诊断价值

目的 探讨原发性肝癌患者血清S100A8和S100A9的表达水平及其与临床病理特征的相关性.方法 收集原发性肝癌患者共90例,同期选择在我院进行体检健康者50例作为对照组;检测2组血清S100A8和S100A9的水平;分析肝癌患者血清S100A8和S100A9水平与临床病理特征的相关性;比较随访期间死亡组与存活组患者血清S100A8和S100A9表达.结果 与对照组比较,肝癌组患者血清S100A8和S100A9均显著升高(P＜0.01).肝癌患者血清S100A8和S100A9的高表达与患者的年龄、性别、大小、病理分型无关(P＞0.05),而与TNM分期、分化程度、淋巴结转移以及血行转移有关(P＜0.01).所有患者6个月定期随访,31例死亡,59例存活;死亡组患者入院时血清S100A8和S100A9表达均显著高于存活组,差异有统计学意义(P＜0.01).结论 原发性肝癌患者血清S100A8和S100A9水平均显著升高,与肝癌的恶性程度关系密切,有望成为原发性肝癌预后的重要评判指标.     

HLA-B gene participates in the NF-kappaB signal pathway partly by regulating S100A8 in the laryngeal carcinoma cell line Hep2

Human leukocyte antigen B (HLA-B), a novel member of the NF-kappaB signal pathway in laryngeal squamous cell carcinoma (LSCC), mediates immunological surveillance of tumor cells by presenting peptides to cytotoxic T-lymphocytes (CTLs) together with S100 calcium binding protein A8 (S100A8). The objective of this study was to investigate the molecular mechanism of HLA-B and S100A8 in laryngeal carcinogenesis. Flow cytometry, 3-(4,5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium bromide (MTT) assay and cell invasion in vitro were used to detect the biological effect of the Hep2 cell line induced by HLA-B RNA interference. RT-PCR and Western blotting were applied to evaluate the expression level of the S100A8 gene after HLA-B RNA interference. Our results showed that HLA-B had negative effects on Hep2 cells by inhibiting apoptosis and cell invasion while decreasing cell proliferation. Additionally, the expression level of HLA-B and S100A8 in LSCC were down-regulated after HLA-B RNA interference. The abnormal expression of HLA-B is thus relevant to the biological effect of laryngeal carcinoma and participates in the NF-kappaB signal pathway partly by regulating the expression of the S100A8 gene.     

High S100A8 and S100A12 protein expression is a favorable prognostic factor for survival of oropharyngeal squamous cell carcinoma

S100/calgranulins (S100A8, S100A9 and S100A12) are key players of innate immune function and elevated levels are a characteristic feature of acute and chronic inflammation, and inflammation‐associated carcinogenesis. However, reduced S100A8 and S100A9 expression has been detected for squamous cell carcinoma, including the head and neck region (HNSCC), which originate from mucosal epithelia with abundant expression of both proteins under physiological conditions. In contrast to S100A8 and S100A9, only sparse information is available for S100A12 and a comparative study of all three S100/calgranulins in HNSCC is still missing. We analyzed S100/calgranulin protein levels in a retrospective patient cohort (n = 131) of oropharyngeal squamous cell carcinoma (OPSCC) by immunohistochemical staining of tissue microarrays. Common characteristics of all three S100/calgranulins were: (i) abundant expression in supra‐basal keratinocytes of normal mucosa with predominant nuclear staining, (ii) low expression in 30.4–51.9% of primary OPSCCs and (iii) variable accumulation of S100/calgranulin‐positive immune cells in the tumor stroma. These features were associated with histopathological characteristics, such as tumor grade, lymph node metastasis and tumor stage. Furthermore, univariate and multivariate analysis revealed worse overall survival of OPSCC patients with simultaneous reduction of S100A8 and S100A12 expression, while expression of S100A9 or presence of the S100A8/S100A9 heterodimer had no impact, suggesting distinct regulation and function of individual S100/calgranulins in the pathogenesis of HNSCCs.     

Distant metastases in laryngeal squamous cell carcinoma

Distant metastases (DM) is the point of concern and seems to be on the rise with the improved control of the laryngeal cancer in the primary site and neck regions. Prognostic factors must be evaluated to improve the detection of DM at early stage of the disease. Therefore, we have analyzed our cases of laryngeal squamous cell carcinoma with DM to find out the risk factors in these patients. We analyzed the records of laryngeal squamous cell carcinoma patients with DM. The records were evaluated according to distant metastases site, TNM staging, the metastases at the neck, treatment and survival. The incidence of DM was 7.2% in our series. Lung is the most common site of DM in laryngeal squamous cell carcinoma. Staging grouping has been helpful in predicting DM, most of the cases were in stage III and IV (85%). Supraglottic lesions is the most common site in patients with DM. The recurrence in the locoregional site was observed in 47% of cases. The overall survival with DM is 28 months, without DM 22 months. The patients with DM in laryngeal squamous cell carcinoma were from the group with supraglottic lesions, stage 3, 4a and 4b. Stage grouping seems to be a better indicator of DM rather than T or N stage alone. The most common site of metastasis is the lung.     

喉鳞状细胞癌中PIN1基因扩增及表达异常

目的：研究PIN1基因在喉癌组织中的表达情况。方法：采用差异PCR、核型分析方法检测喉鳞癌及癌旁对照组织中PIN1基因扩增情况;采用RT-PCR检测同批组织中PIN1 mRNA表达情况;采用免疫组织化学、Western Blot方法检测PIN1蛋白表达情况。结果：与癌旁对照组织相比,40例喉癌组织中有9例（9/40,22.5%）存在PIN1基因扩增,27例（27/40,67.5%）PIN1 mRNA过表达,26例（26/40,65%）PIN1蛋白过表达（P〈0.05）。40例中17例原代培养成功,可制备中期染色体标本,其中3例（3/17,17.65%）存在19号染色体增加。结论：喉鳞状细胞中PIN1基因拷贝数增加,PIN1 mRNA和蛋白质过表达,可能与喉癌发生发展有关。