FTO在胃癌组织中的表达降低及其对细胞系MGC-803功能的影响

目的探讨RNA甲基化相关蛋白FTO在胃癌中的表达变化情况,及其对胃癌细胞功能的影响。方法用realtime PCR检测54例胃癌组织及对应癌旁组织中FTO的表达;用质粒(p CMV6-FTO)实现FTO在胃癌细胞系MGC-803中的过表达;通过增殖实验、划痕试验和Transwell实验,检测FTO对MGC-803细胞功能的影响。结果 FTO mRNA在胃癌组织的表达水平显著低于癌旁组织(P0.05),且其表达与胃癌样本的临床分期密切相关(P0.05);在MGC-803细胞系中过表达FTO,可抑制胃癌细胞增殖(P0.05)、抑制细胞迁移(P0.05)和侵袭(P0.05)。结论 FTO在胃癌组织中低表达,并且抑制胃癌细胞系的增殖、迁移和侵袭,可能与胃癌发生发展密切相关。     

DADS通过miR-222抑制人胃癌细胞系的增殖与侵袭

目的通过miRNA途径研究二烯丙基二硫(DADS)的抑瘤机制,以进一步阐明DADS抑制胃癌细胞增殖与转移的分子机制。方法将胃癌细胞系MGC-803细胞分为DADS处理组、miR-222模拟物组、miR-222抑制物组和阴性对照组;分别采用0、25、50、100、200和400μmol/L DADS处理MGC-803细胞;分别将miR-222模拟物、miR-222抑制物和scramble转染MGC-803细胞。qRT-PCR检测MGC-803细胞中miR-222表达;MTT法和Transwell侵袭实验检测MGC-803细胞增殖与侵袭能力;蛋白质印迹试验检测MGC-803细胞中TIMP3蛋白表达。结果 DADS可呈剂量依赖性下调MGC-803细胞中miR-222表达(P0.05);DADS能抑制MGC-803细胞的增殖与侵袭,外源高表达miR-222能促进MGC-803细胞的增殖与侵袭,而miR-222抑制物与DADS共同处理,MGC-803细胞的增殖与侵袭抑制作用最为显著(P0.05);DADS可下调MGC-803细胞中TIMP3蛋白的表达,外源高表达miR-222能上调MGC-803细胞中TIMP3蛋白的表达,而miR-222抑制物与DADS共同处理,MGC-803细胞中TIMP3蛋白的表达下调最为显著(P0.05)。结论 DADS通过下调miR-222的表达,靶向TIMP3抑制胃癌细胞的增殖与侵袭。     

miR-216a-5p通过抑制HMGB1降低胃癌细胞系MGC-803的增殖和迁移

目的探讨miR-216a-5p对胃癌细胞增殖和迁移的影响及其分子机制。方法用Western blot检测胃癌和癌旁组织中高迁移率族蛋白1(HMGB1)的表达,RT-qPCR检测miR-216a-5p的表达。用人工合成的miR-216a-5p模拟物和其抑制物转染胃癌细胞系MGC-803。MTT检测细胞增殖,划痕法检测细胞迁移。双荧光素酶报告实验和Western blot检测miR-216a-5p和HMGB1之间的靶向关系。结果与癌旁组织相比,HMGB1在胃癌组织中高表达,miR-216a-5p在胃癌组织中低表达(P0.05)。miR-216a-5p模拟物上调胃癌细胞miR-216a-5p的表达水平,抑制细胞的增殖和迁移;miR-216a-5p抑制物下调miR-216a-5p的表达水平,促进胃癌细胞的增殖和迁移(P0.05)。miR-216a-5p可作用于HMGB1的3′UTR并抑制其表达。结论 miR-216a-5p通过靶向抑制HMGB1的表达降低胃癌细胞系MGC-803的增殖和迁移。     

癌基因HuR调节胃癌功能的研究

目的:探讨癌基因HuR在胃癌组织的表达和对胃癌MGC-803细胞功能的影响。方法:使用RT-qPCR分析80例临床确诊的胃癌患者胃癌组织样本中HuR的表达水平;利用pSIH载体构建敲减HuR的重组质粒,实现HuR在MGC-803细胞中的低表达,以空载体pSIH作为对照;使用划痕实验检测细胞迁移能力,使用Transwell实验检测细胞侵袭能力,使用CCK-8法检测细胞活力。结果:RT-qPCR结果显示,与癌旁对照相比,67例(84%)胃癌组织中出现HuR的表达上调;低表达HuR能显著抑制MGC-803细胞的侵袭能力、迁移能力和活力(P0.05),提示HuR作为癌基因起作用。结论:HuR的异常表达可能与胃癌的发生发展有关,低表达HuR可以抑制胃癌MGC-803细胞的侵袭、迁移和活力。     

Mi R-31在胃癌组织中低表达并抑制HGC-27细胞增殖、迁移与侵袭

目的研究miR-31在胃癌患者中的表达,探讨miR-31对胃癌HGC-27细胞增殖、侵袭和转移的影响。方法收集胃癌患者标本39例,同时收取相应的癌旁组织20例。通过荧光定量PCR方法检测各组织中miR-31的表达水平。以脂质体为载体将miR-31模拟物转染至HGC-27细胞,CCK-8方法检测细胞增殖能力变化,Transwell实验检测HGC-27细胞侵袭和迁移能力变化。结果 miR-31在胃癌组织中的表达水平明显低于癌旁组织,P0.01。过表达miR-31能够显著抑制HGC-27细胞的增殖,显著抑制HGC-27细胞的侵袭和迁移能力(P0.01)。结论 MiR-31在胃癌中低表达,并抑制胃癌细胞系HGC-27的增殖、迁移和侵袭。     

胃癌中MCU的表达及对胃癌细胞转移的作用

目的探讨线粒体钙单向转运蛋白(mitochondrial calcium uniporter, MCU)在胃癌组织中的表达以及对胃癌细胞转移的影响。方法采用免疫组化SP法检测60例原发性胃癌和癌旁组织中MCU的表达;采用免疫荧光法和Western blot法检测20例新鲜胃癌组织及癌旁组织中MCU的表达。采用CCK-8实验检测MCU诱导剂(Spermine)对人胃癌细胞(MGC-803)增殖的影响。应用Transwell和细胞划痕实验检测对照组、Spermine组、siRNA MCU组对MGC-803细胞侵袭和迁移能力的影响。根据不同分组,干预48 h,通过线粒体膜电位检测钙离子,Western blot法检测MCU蛋白的表达水平。结果胃癌组织中MCU表达水平显著高于癌旁组织。细胞实验结果显示,Spermine对MGC-803细胞增殖的影响呈剂量依赖性。Spermine显著增加MGC-803细胞迁移和侵袭能力,siRNA MCU组显著降低细胞侵袭和迁移的细胞数量。线粒体膜电位结果表明,MCU调控胃癌细胞线粒体膜电位钙离子的水平。siRNA MCU组显著降低HIF-1α、TGF-β和增加E-cadherin蛋白的表达。结论 MCU在胃癌组织中高表达并促进胃癌细胞侵袭和迁移,对HIF-1α和TGF-β表达具有调控作用。     

miR-195抑制舌鳞癌细胞系CAL27增殖、迁移和侵袭

目的研究miR-195对舌鳞癌CAL27细胞增殖、迁移和侵袭的影响及其机制。方法用RT-qPCR检测舌鳞癌组织及癌旁正常组织中miR-195 mRNA的表达; CAL27细胞的分组及处理:miR-195组(转染miR-195 mimics)、miR-con组(转染miR-NC)、si-con组(转染si-con)、si-激酶12抑制剂SUZ12组(转染si-SUZ12)、miR-195+Ctrl组(miR-195 mimics和pc DNA 3. 1共转染)、miR-195+SUZ12组(miR-195 mimics和pc DNA 3. 1-SUZ12共转染); MTT法检测细胞增殖; Transwell检测细胞迁移和侵袭; Western blot检测细胞SUZ12蛋白表达;双荧光素酶报告基因检测细胞荧光活性。结果舌鳞癌组织中miR-195 mRNA表达与癌旁正常组织及与健康口腔角质细胞NHOK相比显著降低(P0. 05);过表达miR-195及敲减SUZ12均可显著抑制CAL27细胞增殖、迁移及侵袭(P0. 05);过表达SUZ12可逆转miR-195对CAL27细胞增殖、迁移和侵袭的抑制。结论 miR-195可抑制CAL27细胞的增殖、迁移和侵袭,其机制可能与靶向SUZ12有关。     

miR-551b-3p在胃癌组织中低表达并抑制胃癌细胞系HGC-27的增殖、迁移和侵袭

目的探讨miR-551b-3p在胃癌中的表达变化情况,及其对胃癌细胞功能的影响。方法用real-time PCR的方法检测60例胃癌组织及其对应的癌旁组织中miR-551b-3p的表达量,使用miR-551b-3p模拟物(miR-551b-3p mimic)转染胃癌细胞系HGC-27,CCK-8法检测细胞增殖;划痕愈合实验检测细胞迁移;Transwell法检测细胞侵袭。结果 1)miR-551b-3p在胃癌癌症组织中的表达明显低于癌旁组织(P0.05)。2)过表达miR-551b-3p mimic可以明显减弱胃癌细胞系HGC-27的增殖、迁移和侵袭能力。结论 miR-551b-3p在胃癌组织中低表达,并且抑制胃癌细胞系HGC-27的增殖、迁移和侵袭,可能与胃癌发生发展密切相关。     

miR-26a下调COX-2的表达抑制胃癌AGS细胞增殖和侵袭

目的探讨miR-26a在胃癌组织和胃癌AGS细胞中表达量的改变及其对胃癌AGS细胞增殖和侵袭的影响。方法采用Real-time PCR法检测18例患者胃癌组织及对应的癌旁组织中miR-26a的表达。胃癌AGS细胞分别转染miR-NC和miR-26a,采用Western blot法检测转染前后细胞中COX-2的表达水平,同时采用CCK-8和克隆形成实验检测转染后AGS细胞的增殖和侵袭情况。结果 miR-26a在胃癌组织中的表达明显低于癌旁组织(P0.01);COX-2在乳腺癌组织和细胞中的表达明显高于癌旁组织(P0.01)。转染miR-26a后,胃癌AGS细胞内COX-2的表达量显著下调。CCK-8和克隆形成实验结果显示,过表达miR-26a能明显抑制胃癌AGS细胞的增殖和侵袭(P0.01)。结论 miR-26a抑制胃癌AGS细胞的增殖和侵袭可能与下调COX-2的表达相关。     

hsa-miR-21-5p/ZNF367分子轴通过PI3K/Akt通路影响胃癌细胞的增殖和迁移

目的探讨ZNF367通过PI3K/Akt通路抑制胃癌(GC)细胞的增殖和迁移,并初步探讨其作用分子机制。方法收集2015年8月–2018年10月上海市第八人民医院普外科手术切除的GC组织(非坏死部分)和相邻癌旁组织(距肿瘤组织3 cm)标本46例,同时选取正常人胃黏膜上皮细胞GES1及GC细胞系MGC-803和MKN-45。采用qRT-PCR、Western blot实验检测ZNF367和hsa-miR-21-5p(mi R-21-5p)在组织和细胞系中的表达情况,进行Kaplan-Meier生存分析得出总生存期;利用脂质体LipofectamineTM 2000转染寡核苷酸片段mi RNA-NC、miR-21-5pmimic和miR-21-5pinhibitor至MGC-803细胞,通过CCK8法和划痕实验检测细胞的增殖和迁移能力;采用双荧光素酶报告载体系统验证ZNF367和miR-21-5p的作用靶点,采用qRT-PCR、 Westernblot实验检测miR-21-5pmimic和miR-21-5pinhibitor转染MGC-803细胞中ZNF367、E-cadherin、N-cadherin、Vimentin以及PI3K/Akt通路中p-PI3K和p-Akt的表达情况。结果 ZNF367在胃癌组织和胃癌细胞系中的表达水平分别显著低于癌旁组织和GES-1细胞(P0.01),且在MGC-803细胞中表达水平最低,而miR-21-5p的表达正相反;在MGC-803细胞中,OE-ZNF367和miR-21-5p inhibitor转染组的细胞增殖和迁移能力显著低于对照组,而si RNA-ZNF367和miR-21-5pmimic转染组的细胞增殖和迁移能力显著高于对照组;生物信息学和荧光素酶活性实验结果表明ZNF367与miR-21-5p具有直接靶向关系;qRT-PCR、Western blot实验结果表明在miR-21-5p inhibitor转染组中,E-cadherin的表达是显著提高的(P 0.01),而N-cadherin、Vimentin、p-PI3K和p-Akt的表达是显著下降的(P0.01);在miR-21-5pmimic转染组中,结果相反(P 0.01)。结论 hsa-miR-21-5p/ZNF367分子轴通过调控PI3K/Akt通路抑制EMT过程,从而影响胃癌细胞MGC-803的增殖和迁移。     

Clear cell basal cell carcinoma

We describe a case of clear cell basal cell carcinoma of the superficial type, presenting as a crusted eruption on the abdomen. Histological examination showed a solid proliferation of clear cells attached to the under-surface of an atrophied epidermis. In addition, distinct pagetoid infiltration was seen within the overlying epidermis. A focal connection between the clear cell portion and a deeper lying nodular basal cell carcinoma was demonstrated, elucidating the true nature of the lesion. Immunohistochemical studies and electronmicroscopy confirmed the epithelial derivation of the tumour. The clear cell appearance was due to multiple cytoplasmic electronlucent vacuoles which were not surrounded by membranes.     

Absolute dependence of T cell receptorhi cell generation and relative dependence of T cell receptorint cell generation on the thymus

Recent evidence indicates that conventional T cells are generated by the mainstream of T cell differentiation in the thymus and acquire a high density of T cell receptor expression (i.e. TCR^hi). In contrast, primordial T cells (or NK1.1⁺ T cells) are generated by the extrathymic pathways or an alternative intrathymic pathway and express an intermediate density of TCR (i.e. TCR^int). To obtain further evidence, it was examined how thymus grafting influenced the distribution of T cell populations in athymic nude mice. When BALB/c nu/nu mice were engrafted with thymocyte-depleted BALB/c+/+ fetal thymi, two changes emerged after grafting: nude mice generated TCR^hi cells de novo in the periphery as well as in the grafted thymi, and the absolute number of interleukin-2 receptor β chain⁺ TCR^int cells increased prominently in number in the periphery. Among thymic hormones tested, the administration of thymosin α induced a slight expansion of CD3^int cells in nude mice. To examine a possible interaction of TCR^int cells with TCR^hi cells in the periphery, B6 nu/nu mice (Ly5.2⁺) were injected with TCR^hi cells purified from the spleen of B6 Ly5.1 congenic mice. In this case, TCR^int (Ly5.2⁺) cells expanded well in all tested organs of nude mice. These results suggest that the generation of TCR^hi cells is absolutely dependent on the thymus and that TCR^int cells expand under the influence of the thymus (humoral) and due to interaction with thymus-derived conventional T cells.     

树突状细胞对调节性T细胞的调控作用

树突状细胞(DCs)是目前已知的体内功能最强大的专职性抗原提呈细胞,具有启动免疫应答和诱导免疫耐受的双重特性.近年来树突状细胞对调节性T细胞的调控作用是免疫学领域的一个研究热点.Foxp3+ Tregs是一群同时具有免疫低反应性和免疫抑制性功能两大特征的T淋巴细胞,它在维持机体内环境稳定、预防自身免疫性疾病、抑制移植排斥反应等病理生理过程中发挥着重要作用.越来越多的研究结果证实DCs和Tregs二者在维持外周免疫耐受中存在着紧密联系,DCs可以诱导抗原特异性Tregs的生成并增加后者的抑制活性,其中参与该调节机制的分子主要包括相关细胞因子、Toll样受体、共刺激分子及维甲酸等.对DCs在接触共生和致病微生物时诱导和调控Tregs细胞有了一些新发现.     

具有透明细胞乳头状肾细胞癌形态特征的透明细胞性肾细胞癌的临床病理特征

目的探讨具有透明细胞乳头状肾细胞癌形态特征的透明细胞性肾细胞癌(clear cell papillary renal cell carcinoma-like clear cell renal carcinoma, CCPRCC-like CCRCC)的临床和病理特征, 旨在提高对该类肿瘤的认识。方法回顾性分析浙江大学医学院附属第一医院诊断的3例CCPRCC-like CCRCC的临床资料、组织学形态及免疫表型, 随访患者预后并复习相关文献。结果男性2例, 女性1例, 平均年龄43岁。肿块最大径9 cm。镜下观察3例病例均包含透明细胞乳头状肾细胞癌样(CCPRCC-like)及透明细胞性肾细胞癌样(CCRCC-like)两种区域, 前者在整个瘤体所占比例为20%～90%。CCPRCC-like区域含有囊、乳头、腺管状结构。细胞核远离基底膜, 细胞核WHO/国际泌尿病理协会(ISUP)分级1级。在CCPRCC-like区域, 细胞角蛋白(CK)7表达范围10%～80%, 中等及以上强度表达。CD10表达均为100%, 主要为顶端胞膜表达。在CCRCC-like区域, 1例CK7阴性, 其余...     

梭形细胞间变性大细胞淋巴瘤

1.病例简介:患者男,40岁.2个月前自觉右腹股沟区疼痛,行走时加剧,于2009年6月在外院行B超检查发现腹股沟区多枚淋巴结肿大,直径约4.0～5.0 cm,胰周、腹主动脉及髂血管旁多发低回声实性结节,最大直径为4.5 cm;行CT检查,提示腹膜后、肝门部多发肿大淋巴结.血常规检查:白细胞、中性粒细胞升高,淋巴细胞减少.2009年7月23日手术切除右腹股沟淋巴结.原单位病理诊断:考虑为指状突树突状细胞肿瘤.因多家医院诊断结果存在分歧,遂来我院会诊,详细询问病史,患者既往无其他脏器肿瘤病史.     

Galectins: versatile modulators of cell adhesion, cell proliferation, and cell death

 Lectins, or carbohydrate binding proteins, recognize specific oligosaccharide structures on glycoproteins and glycolipids. Several families of animal lectins have been identified; for some of these lectins, functions such as leukocyte adhesion and microbial opsonization have been described. The galectins are a family of lectins found in species ranging from sponges and nematodes to humans. Members of the galectin family have been proposed to mediate cell adhesion, to regulate cell growth, and to trigger or inhibit apoptosis. The expression pattern of different galectins changes during development, and this pattern is also altered at sites of inflammation and in breast, colon, prostate, and thyroid carcinomas. In addition, the level of expression of some galectins by tumor cells has been shown to be correlated with metastatic potential. The mechanisms by which galectins exert these diverse effects remain largely unknown. Some glycoprotein counterreceptors recognized by certain galectins have been identified; this is an important first step in understanding the cell-type specific effects of different galectins. This review discusses the way in which the modulation of galectin activity may affect strategies for treatment of a variety of human diseases, including autoimmunity and cancer. Received: 4 April 1997 / Accepted: 24 September 1997     

Between-observer repeatability of sputum differential cell counts. Influence of cell viability and squamous cell contamination

BACKGROUND: Induced sputum differential cell counts have been advocated as a method of noninvasively assessing airway inflammation in asthma and other airway diseases. Relatively little is known about the between-observer repeatability of sputum differential cell counts and the factors that influence it. OBJECTIVE: To assess the between-observer variability of induced sputum cell counts. METHODS: Sputum was induced and processed using standard techniques. Forty-two slides from 38 patients (31 with asthma, seven normal subjects) were randomly selected. Slides were classified as good (<20% squamous cells and >50% viability; n = 24); low viability (<50% viability; n = 10) and high squamous cell contamination (>20% squamous cells; n = 8). Two blinded observers counted between 200 and 400 nonsquamous cells and agreement was assessed by the intraclass correlation coefficient (ICC) and the standard deviation of between-observer differences (SD). RESULTS: The overall ICC were 0.9, 0.89, 0.9 for eosinophils, neutrophils and macrophages and 0.29 and 0.69 for lymphocytes and epithelial cells. Repeatability was greater in slides classified as good compared with slides with low cell viability and particularly excess squamous cell contamination. CONCLUSIONS: We have shown that the overall between-observer repeatability of the differential eosinophil, neutrophil and macrophage cell counts is good. Low cell viability and particularly excess squamous cell contamination reduce between-observer repeatability suggesting that techniques that ensure high cell viability and reduce squamous contamination would be an advantage.