沉默LIMK1抑制人结肠癌SW480细胞上皮-间质转化

目的:研究沉默LIMK1对人结肠癌SW480细胞上皮-间质转化（epithelial-mesenchymal transformation，EMT）的影响。方法:RT-PCR、Western blot、免疫荧光与免疫组化检测沉默LIMK1对EMT相关分子表达的影响。裸鼠实验检测沉默LIMK1对移植瘤生长的影响。结果:RT-PCR检测显示，沉默LIMK1可下调Vimentin和上调E-cadherin mRNA表达(P<0.05)。Western blot显示，沉默LIMK1可明显下调SW480细胞Snail及Vimentin蛋白表达和上调E-cadherin表达(P<0.05)。免疫荧光显示，LIMK1沉默细胞Snail与Vimentin阳性信号较对照组明显减弱，而E-cadherin阳性信号显著增强。裸鼠实验结果显示，沉默组肿瘤生长速度较SW480组明显减慢(P<0.05)。沉默组移植瘤瘤重（0.16±0.079）g较SW480组（0.86±0.165）g明显减少，瘤重抑制率为97.7%(P<0.05)。LIMK1沉默组移植瘤较对照组的Ki-67、Vimentin、Snail与CD34阳性表达明显减弱，而E-cadherin表达明显增加。结论:沉默LIMK1可体外抑制人结肠癌SW480细胞EMT。     

Akt/GSK-3β/Snail通路在TGF-β_1诱导A549/DDP细胞上皮间质转化中的作用

目的:通过观察转化生长因子β_1(TGF-β_1)诱导前后Akt/GSK-3β/Snail信号通路相关分子的表达情况,探讨A549/DDP细胞发生上皮-间充质转化(epithelial-mesenchymal transition,EMT)的分子机制。方法:TGF-β_1(5μg/L)作用48 h,观察A549/DDP细胞的形态变化,并测定EMT标志物上皮型钙黏蛋白(E-cadherin)和神经型钙黏蛋白(N-cadherin)的表达情况,评估TGF-β_1是否成功诱导A549/DDP细胞发生EMT。进一步将A549/DDP细胞分为TGF-β_1(+)组、TGF-β_1(-)组和LY294002组,应用Western blot法检测各组Akt/GSK-3β/Snail通路相关分子Akt、p-Akt、GSK-3β、p-GSK-3βSer9和Snail的蛋白水平。结果:形态学观察可见TGF-β_1(+)组的细胞变得更加狭长,呈纺锤形,细胞间较为疏散,形态与间充质细胞相似。与TGF-β_1(-)组比较,TGF-β_1(+)组E-cadherin蛋白表达下调,N-cadherin蛋白表达上调(P0.05);各组间Akt和GSK-3β的蛋白表达水平并无明显差别,TGF-β_1(+)组的p-Akt、p-GSK-3βSer9和Snail蛋白水平较TGF-β_1(-)组明显增强(P0.05);PI3K抑制剂LY294002与TGF-β_1同时刺激细胞后,p-Akt、p-GSK-3βSer9和Snail的蛋白水平较TGF-β_1(+)组下调(P0.05)。结论:TGF-β_1干预Akt/GSK-3β/Snail信号通路,促进Akt和GSK-3β的磷酸化,提高Snail的表达水平,诱导A549/DDP细胞发生EMT。     

转录抑制因子Snail表达与胃癌Lauren分型的关系研究

目的探讨转录抑制因子Snail表达与胃癌Lauren分型的关系。方法Western印迹检测肠型胃癌细胞系（N87）和弥漫型胃癌细胞系（AGS）中Snail和上皮型E钙黏蛋白（E-cadherin）的表达。将糖原合酶激酶．3B（GSK-3B）质粒转染胃癌细胞系,检测Snail和E-cadherin的表达变化。收集2000年2月至2005年12月间在复旦大学附属中山医院行胃癌根治术的77例术后组织标本．采用免疫组织化学染色检测Snail在胃癌组织中表达．并分析其与胃癌Lauren分型之间的关系。结果Snail在N87中低表达,在AGS中高表达;E-cadherin表达与Snail相反。转染GSK-3B后,胃癌细胞Snail表达显著下调,E-cadherin表达显著上调（P〈0．01）。使用不同浓度的GSK-3B抑制剂氯化锂处理后．胃癌细胞Snail表达显著上调,且具有明显浓度依赖性（P〈0．01）。21例肠型胃癌中Snail低表达16例,高表达5例;56例弥漫型胃癌中Snail低表达21例,高表达35例;肠型胃癌中Snail表达明显弱于弥漫型,差异有统计学意义（P〈0．01）。结论Snail的表达与胃癌Lauren分型有关．是一种潜在的确定胃癌分型的分子标志物。     

Snail与 E－cadherin在口腔鳞癌中的表达及临床意义

目的：研究口腔鳞癌中转录因子Snail和黏附分子E－cadherin表达,并探索二者与口腔鳞癌进展转移关系。方法应用免疫组化的方法,研究38例口腔鳞癌组织中Snail与E－cadherin的表达情况,并与各项临床病理参数进行对照分析。结果口腔鳞癌组织Snail的表达程度随组织分化程度的降低而逐渐增强,伴有淋巴结者高表达;E－cadherin的表达程度随组织分化程度的降低而逐渐降低,伴有淋巴结转移者低表达。结论（1）Snail和E－cadherin蛋白的表达与口腔鳞癌恶性程度相关。（2）Snail蛋白可能在转录水平上调控E－cadher in蛋白的表达。     

Synergistic effects of snail and quercetin on renal cell carcinoma Caki-2 by altering AKT/mTOR/ERK1/2 signaling pathways

Renal cell carcinoma has become the most common subtype of kidney cancer, and has the highest propensity to manifest as metastatic disease. Because of lack of knowledge in events that correlated with tumor cell migration and invasion, few therapeutic options are available. Therefore, in current study, we explore the anti-tumoral effect of a potential chemopreventive natural product, quercetin, combined with anti-sense oligo gene therapy (inhibiting Snail gene). We found that either one of them had the remarkable effects in suppressing cell proliferation and migration, inducing cell cycle arrest and apoptosis in a ccRCC cell line, Caki-2 cells. The combination of both means provides even strong suppressive effects toward these ccRCC cells. Our study, for the first time, provides the possibility of using a novel treatment for renal cancer, by combining natural product and gene therapy.     

辣椒素对人大细胞肺癌NCI-H460细胞侵袭能力及E-钙黏蛋白、Snail蛋白表达的影响

目的观察辣椒素对人大细胞肺癌NCI-H460细胞侵袭能力及E-钙黏蛋白(E-cadherin)和Snail蛋白表达的影响,探讨其抗非小细胞肺癌的可能机制。方法体外培养NCI-H460细胞,以不同终浓度辣椒素进行处理,未加辣椒素为对照组。采用Hoechst33342核染色分析、Transwell小室细胞侵袭实验及Western blot检测,观察辣椒素对NCI-H460细胞凋亡、侵袭能力及E-cadherin、Snail蛋白表达变化的影响。结果与对照组比较,Hoechst33342核染色分析表明,辣椒素各浓度组可明显诱导NCI-H460细胞凋亡(P0.05);Transwell体外侵袭实验结果显示,辣椒素可显著抑制侵袭穿膜细胞数(P0.05);Western blot检测结果表明,辣椒素各浓度组E-cadherin表达水平明显升高、Snail表达水平明显降低(P0.05)。结论辣椒素能明显诱导NCI-H460细胞凋亡;降低Snail表达、刺激E-cadherin的表达从而抑制NCI-H460细胞侵袭能力,可能是其抗非小细胞肺癌的机制之一。     

Cisplatin-selected resistance is associated with increased motility and stem-like properties via activation of STAT3/Snail axis in atypical teratoid/rhabdoid tumor cells

Atypical teratoid/rhabdoid tumor (ATRT) is a malignant pediatric brain tumor with great recurrence after complete surgery and chemotherapy. Here, we demonstrate that cisplatin treatment selects not only for resistance but also for a more oncogenic phenotype characterized by high self-renewal and invasive capabilities. These phenomena are likely due to STAT3 upregulatoin which occurred simultaneously with higher expression of Snail, an activator of epithelial–mesenchymal transition (EMT), in ATRT-CisR cells. STAT3 knockdown effectively suppressed Snail expression and blocked motility and invasion in ATRT-CisR cells, while overexpressing Snail reversed these effects. Chromatin immunoprecipitation assay indicated that STAT3 directly bound to Snail promoter. Moreover, STAT3 knockdown effectively suppressed cancer stem-like properties, synergistically enhanced the chemotherapeutic effect, and significantly improved survival rate in ATRT-CisR-transplanted immunocompromised mice. Finally, immunohistochemistrical analysis showed that STAT3 and Snail were coexpressed at high levels in recurrent ATRT tissues. Thus, the STAT3/Snail pathway plays an important role in oncogenic resistance, rendering cells not only drug-resistant but also increasingly oncogenic (invasion, EMT and recurrence). Therefore, the STAT3/Snail could be a target for ATRT treatment.     

湖沼地区钉螺指数的初步研究

目的利用遥感与地理信息系统(GIS)技术自动确定钉螺孳生地。方法  利用丰水期与枯水期遥感图像，分别自动计算与提取水体指数(NDWI)，两图像水体指数相减，获得“冬陆夏水”区域；在“冬陆夏水”区域计算植被指数(NDVI)，并提取有植被覆盖的区域；将提取的区域与已有螺情资料进行空间叠加对比分析。结果在鄱阳湖区，两时相中NDWI>0. 05的区域之差与NDVI>0. 20的区域的交集，为适宜钉螺孳生的区域。结论根据适宜钉螺孳生的生态条件，利用遥感与GIS技术可以快速获得钉螺的孳生地及其面积。         

常州市有螺环境现状调查分析

目的 调查常州市有螺环境改造现状,为螺情监测提供依据.方法 收集全市有螺环境资料,对所有有螺环境进行现场调查并进行环境分类,建立数据库分析各类环境特点,并应用Google Earth软件绘制分布图.结果 常州市有螺环境共计6 174个,面积3 911.97万m2,分布于7市(县、区)的51个乡(镇)、414个村.根据环境分类,Ⅱ类环境最多,Ⅰ、Ⅱ、Ⅲ、Ⅳ类环境面积的构成比分别为24.8％、54.4％、11.3％、9.5％.金坛市、武进区、溧阳市的有螺环境面积最大,分别占全市总面积的31.0％、24.4％、24.0％.金坛市、溧阳市、武进区、新北区以现况Ⅱ类环境居多,天宁区、钟楼区、戚墅堰区以现况Ⅰ类环境居多.Ⅳ类环境主要分布在武进区和新北区,分别占68.3％、25.9％.江湖滩、内陆和山丘3种类型流行区有螺环境面积的构成比分别为9.9％、49.8％、40.3％.3种类型流行区均以Ⅱ类环境居多,面积分别占52.3％、50.1％、60.2％,Ⅳ类环境所占比例分别为22.5％、14.1％、0.7％.结论 常州市有螺环境分布广,面积大,应重点加强对Ⅲ类、ⅣV类环境的监测.