结直肠癌PTEN基因表达及与p-ERK相关性研究

目的 研究结直肠癌中的PTEN、p-ERK蛋白的表达及相互关系,初步探讨它们在结直肠癌的发生发展中的生物学意义.方法 应用免疫组织化学染色快捷法,检测40例结直肠癌组织、18例结直肠腺瘤、13例结直肠正常黏膜中PTEN蛋白、和p-ERK蛋白的表达情况,比较PTEN蛋白表达与临床病理指标的关系,及其与p-ERK蛋白表达的相关性.结果 1.结直肠癌癌组织PTEN蛋白表达的阳性率(57.5％)明显弱于腺瘤(72.2％)及正常组织(100％),组间比较差异有显著性(P＜0.05),其表达水平与结直肠癌的分化程度、淋巴结转移、浸润深度、Dukes分期有关,与患者的性别、年龄,肿瘤大小及位置无关(P＞0.05)2.结直肠癌组织p-ERK蛋白表达的阳性率(72.5％)明显高于正常结直肠黏膜组(0.00％)及腺瘤组(66.6％),组间比较差异有显著性(P＜0.01),其表达随结直肠癌侵润深度增加、淋巴结转移、Duke分期的进展而增高.3.PTEN蛋白表达强度与p-ERK蛋白表达强度之间呈负相关(r=-0.452,P＜0.05).结论 提示抑癌基因PTEN的表达与结直肠癌生物学行为密切相关;在结直肠癌发生、发展过程中,可能由于PTEN蛋白的低表达或失表达不能有效抑制ERK磷酸化,使细胞发生癌变,并促进癌变细胞的浸润、转移.     

组织微阵列技术研究肿瘤转移相关基因在鼻咽癌中的表达与临床意义

利用高通量组织微阵列结合免疫组化检测MT1-MMP、MT2-MMP、Ezrin、nm23-H1、E-cad和TIMP-2在鼻咽癌组织中的蛋白质表达,探讨肿瘤转移相关基因异常表达在鼻咽癌侵袭转移中的作用,筛选鼻咽癌转移相关分子标志物.结果发现,鼻咽癌组织存在MT1-MMP、Ezrin蛋白高表达(P<0.01)和nm23-H1、TIMP-2蛋白低表达(P<0.05).临床Ⅱ期、Ⅲ期和Ⅳ期鼻咽癌和淋巴结转移鼻咽癌中MT1-MMP、MT2-MMP和Ezrin蛋白阳性表达显著高于临床Ⅰ期鼻咽癌和无转移癌(P<0.05,P<0.01),但临床Ⅱ期、Ⅲ期和Ⅳ期鼻咽癌和淋巴结转移鼻咽癌中nm23-H1蛋白的阳性表达显著低于临床Ⅰ期鼻咽癌和无转移癌(P<0.05).鼻咽癌组织中MT1-MMP与MT2-MMP r=0.308,P<0.001),nm23-H1与E-cad(r=0.167,P<0.05)及TIMP-2(r=0.279,P=0.001),E-cad与TIMP-2(r=0.279,P=0.001)的蛋白质表达旱显著正相关.MT1-MMP与E-cad(r=-0.188,P<0.05)及TIMP-2(r=-0.233,P<0.05),Ezrin与E-cad(r=-0.204,P<0.05)的蛋白质表达呈显著性负相关.聚类分析显示,鼻咽癌MT1-MMP、MT2-MMP和Ezrin蛋白共同阳性表达显著高于慢性炎性鼻咽上皮(P<0.05),但nm23-H1、E-cad和TIMP-2蛋白在鼻咽癌组织中的共同阴性显著高于癌旁上皮和慢性炎性鼻咽上皮(P<0.05,P<0.01).多因素分析和有效性评估发现,MT1-MMP蛋白能较好地独立预测鼻咽癌淋巴结转移和临床进展.上述研究结果提示,多个肿瘤转移基因的蛋白质高表达,转移抑制基因的低表达和这些基因的蛋白质表达失平衡在鼻咽癌淋巴结转移和临床进展过程中起重要作用.MTI-MMP蛋白可作为预测鼻咽癌淋巴结转移的较好分子标志.     

胃癌及相应癌旁组织中VEGF、CD34、CD44v6的检测及临床意义

目的:探讨血管内皮生长因子(Vascular endothelial growth factor,VEGF)、CD34和CD44v6在胃癌及相应癌旁组织中的表达及其与临床病理意义.方法:应用免疫组化技术检测60例胃癌以及相应癌旁组织中VEGF、CD34和CD44v6的表达.结果:胃癌组织VEGF阳性率为71.67%(43/60)明显低于癌旁组织88.34%(53/60),两组间有显著性差异(P=0.022);VEGF阳性表达与胃癌病理分级、浸润深度、淋巴结转移和临床分期密切相关(P＜0.05).癌旁组织中微血管密度(MVD)明显高于癌组织MVD(P=0.000),有淋巴结转移癌组织中MVD高于无淋巴结转移者(P=0.043),并随浸润深度、临床分期MVD升高(P=0.046,P=0.000).癌旁组织中CD44v6阳性率为48.34%(29/60)低于癌组织的61.67%(37/60),两者无明显差别.CD44v6的阳性率随胃癌浸润的加深而升高,与淋巴结转移和TNM分期呈正相关.VEGF、CD34及CD44v6三指标间两两相关(P＜0.05).结论:VEGF、CD34和CD44v6三者联合检测有助于判断胃癌的浸润、转移及预后情况.     

结直肠癌组织BCL9和VEGF的表达与临床意义

探讨结直肠癌中B细胞淋巴瘤因子9(B-cell lymphoma 9,BCL9)和血管内皮生长因子(vascular endothelial growth factor,VEGF)表达与临床病理的关系及其临床意义.采用免疫组织化学SABC染色法,检测83例结直肠癌组织和10例癌旁正常组织中BCL9及VEGF蛋白的表达情况,并且分析其与临床病理特征的关系.BCL9和VEGF在癌组织中表达率分别为67.47%(56/83)和69.88%(58/83),而癌旁正常组织中均不表达.BCL9在结直肠癌中的表达与肿瘤分化程度、浸润深度、Dukes分期、有无淋巴结转移相关(P0.05).VEGF在结直肠癌中的表达与肿瘤分化程度、浸润深度无关,而与肿瘤Dukes分期、有无淋巴结转移密切相关(P0.01).结直肠癌中VEGF的表达随BCL9表达的升高而升高(P0.01).BCL9、VEGF可能在结肠癌的发生发展过程中起重要作用.BCL9可能通过上调VEGF的表达,促进肿瘤浸润和转移.     

胃癌组织中COX-2和nm23的表达及意义

目的:探讨环氧合酶-2(COX-2)和nm23蛋白在胃癌组织中的表达及意义.方法:应用免疫组化方法检测了52胃癌组织及20正常胃黏膜中COX-2和nm23的表达,并在显微镜下计数阳性细胞数,统计比较胃癌组织和正常组织中COX-2和nm23的阳性表达率.结果:COX-2蛋白在胃癌组织和正常胃黏膜组织中阳性表达率分别为67.3%和5.0%,胃癌组织中COX-2蛋白阳性率显著高于正常胃黏膜(P＜0.01).nm23蛋白在胃癌组织和正常胃黏膜组织中阳性表达率分别为30.7%和75.0%,胃癌组织中nm23蛋白阳性率显著低于正常胃黏膜(P＜0.01).COX-2的异常表达与肿瘤细胞分化程度、肿瘤浸润深度、淋巴结转移、TNM分期显著相关(P＜0.05).nm23阳性表达率与肿瘤浸润深度、淋巴结转移、TNM分期显著相关(P＜0.05),而与肿瘤细胞分化程度无显著性相关(P＞0.05).结论:COX-2与nm23表达与胃癌的临床病理特征密切相关.COX-2与nm23可作为反映胃癌侵袭转移、判断预后的生物学指标.     

Snail基因及其蛋白、E-cadherin蛋白在胃癌中的表达及其临床意义

目的观察Snail mRNA及其蛋白、E-cadherin蛋白在胃癌组织中的表达及其与胃癌临床病理特征的关系,并探讨它们在胃癌发生、发展中的作用及其临床应用价值。方法收集96例手术切除胃癌标本,同时取80例癌旁组织作为对照。应用免疫组织化学S-P法检测胃癌组织、癌旁组织中snail蛋白、E-cadherin蛋白的表达;运用原位分子杂交技术检测胃癌组织、癌旁组织中Snail mRNA的表达。结果（1）Snail蛋白在胃癌组织阳性率（83.3%）显著高于癌旁组织（41.25%）（P〈0.05）;高、中分化组Snail蛋白阳性表达率显著低于低分化组（P〈0.05）;Snail蛋白的阳性表达率在乳头状腺癌、管状腺癌及低分化腺癌与黏液癌之间差异有显著性（P〈0.05）;Snail蛋白的表达与胃癌浸润深度、淋巴结转移及远处转移有关（P〈0.05）,与性别、年龄、肿瘤大小、肿瘤部位及临床分期无关（P〉0.05）;（2）胃癌组织中snail mR-NA的阳性率（76%）显著高于癌旁组织（30%）（P〈0.05）;高、中分化组Snail mRNA阳性表达率显著低于低分化组（P〈0.05）;Snail mRNA的阳性表达率在乳头状腺癌、管状腺癌及低分化腺癌与黏液癌之间差异有显著性（P〈0.05）;Snail mRNA的表达与浸润深度及淋巴结转移有关（P〈0.05）,与性别、年龄、肿瘤大小、肿瘤部位、临床分期及远处转移无关（P〉0.05）;（3）E-cadherin蛋白在胃癌组织阳性率（37.5%）显著低于癌旁组织（100%）（P〈0.05）;高、中分化组E-cadherin蛋白阳性率显著高于低分化组（P〈0.05）;E-cadherin蛋白阳性率在乳头状腺癌、管状腺癌及低分化腺癌与黏液癌之间差异有显著性（P〈0.05）;E-cadherin蛋白的表达与胃癌浸润深度、淋巴结转移、临床分期及远处转移有关（P〈0.05）,与性别、年龄、肿瘤大小、肿瘤部位均无关（P〉0.05）;（4）胃癌组织中snail mRNA和snail蛋白的表达呈正相关（r=0.594,P〈0.05）;Snail蛋白和E-cadherin蛋白的表达呈负相关（r=-0.234,P〈0.05）。结论（1）E-cadher-in蛋白低表达与Snail蛋白高表达可能是胃黏膜恶性转变以及胃癌发生浸润转移的重要生物学标志;联合检测E-cadherin蛋白与Snail蛋白对预测胃癌浸润转移有重要意义。（2）Snail蛋白可能在转录水平上调控E-cadherin蛋白的表达。     

结直肠癌COX-2表达及其与临床病理特征的相关性分析

目的:探讨结直肠癌环氧化酶-2(cyclooxygenase-2,COX-2)的表达及其与临床病理特征的相关性。方法:选择2017年8月～2019年6月在本院外科手术诊治的结直肠癌患者60例,取所有患者的病灶组织标本与癌旁组织标本,采用PCR与免疫组化法检测COX-2 mRNA与蛋白表达情况,分析其与患者的临床病理特征的相关性。结果:直肠癌组织COX-2 mRNA与蛋白表达阳性率分别为63.3%和55.0%,显著高于癌旁组织的20.0%和16.7%(P0.05)。随着结直肠癌的病理分期及分化程度的增加、淋巴结转移的发生,直肠癌组织的COX-2 mRNA、蛋白表达阳性率显著升高(P0.05)。Spearman等级相关分析显示直肠癌组织的COX-2 mRNA、蛋白表达阳性率与临床分期、组织学分化与淋巴结转移都存在显著相关性(P0.05)。Cox模型多因素分析显示临床分期、组织学分化与淋巴结转移都是影响COX-2蛋白表达的主要因素(P0.05)。结论:结直肠癌COX-2的mRNA与蛋白都呈现高表达,与患者的临床分期、组织学分化与淋巴结转移显著相关。     

CCNB2在结直肠癌中的表达及临床意义

目的:探讨细胞周期蛋白B2(Cyclin B2,CCNB2)在结直肠癌组织中的表达及其临床意义。方法:选择45对结直肠癌组织及癌旁正常结直肠组织样本,分别采用实时定量PCR(qRT-PCR)方法和免疫组织化学技术检测CCNB2的mRNA和蛋白表达,并进一步分析CCNB2的表达与结直肠癌临床病理特征之间的关系。结果:结直肠癌组织中CCNB2 mRNA的表达显著高于癌旁正常结直肠组织,差异有统计学意义(P0.001),且CCNB2的mRNA表达与结直肠癌的肿瘤大小、浸润深度及TNM分期显著相关(P0.05),与年龄、性别、肿瘤位置、分化程度、脉管神经浸润、淋巴结转移和远处转移均无关(P0.05)。45例结直肠癌标本中39例表达(+~+++),6例表达(-)。CCNB2蛋白主要表达于结直肠癌细胞质中,少量见于细胞核。结直肠癌组织中CCNB2蛋白的阳性表达率为86.7%,显著高于癌旁正常结直肠组织,并与患者的性别、年龄、分化程度和肿瘤转移均无显著相关性(P0.05),但与肿瘤分期、浸润程度均显著相关(P0.05)。结论:CCNB2在结直肠癌中呈异常高表达,且与结直肠癌的发生发展相关,有望作为结直肠癌的诊断和预后预测参考指标。     

Med1/TRAP220和uPAR在非小细胞肺癌中表达的临床意义

目的:探讨Med1/TRAP220和uPAR在非小细胞肺癌中表达的临床病理意义.方法:用免疫组织化学技术检测120例非小细胞肺癌组织中Med1/TRAP220和uPAR的表达,并分析其与患者临床病理指标的关系及其和预后的关系.结果:免疫组化结果显示在120例非小细胞肺癌中Med1/TRAP220的阳性表达率为53.3％,uPAR的阳性表达率为70％,癌旁正常肺组织uPAR阳性表达率为12.5％(15/120),Med1/TRAP220的阳性表达率为80％(96/120).uPAR在肺癌中的阳性表达率显著高于癌旁正常肺组织(P＜0.01),Med1/TRAP220在癌旁正常肺组织中的阳性表达率显著高于肺癌组织(P＜0.01).Med1/TRAP220和uPAR在肺癌中的表达与性别,年龄,肿瘤大小无关(P＞0.05).有淋巴结转移组uPAR的阳性表达率显著高于无淋巴结转移组(P＜0.05),而无淋巴结转移组Med1/TRAP220的阳性表达率显著高于有淋巴结转移组(P＜0.05).uPAR的阳性表达率与非小细胞肺癌的临床分期无显著相关(P＞0.05),而Ⅰ-Ⅱ期非小细胞肺癌组Med1/TRAP220的阳性表达率显著高于Ⅲ-Ⅳ期组(P＜0.05).uPAR在肺鳞癌和腺癌中的表达率有显著差异(P＜0.05),而Med1/TRAP220在肺鳞癌和腺癌中的表达率无显著差异(P＞0.05).uPAR阳性组生存率显著低于uPAR阴性组(P＜0.05),而Med1/TRAP220阳性组生存率显著高于Med1/TRAP220阴性组(P＜0.05).Cox比例危险度模型显示uPAR阳性表达反映不良预后的危险性最大,风险度exp(β)为2.012(P＜0.01),其次为肿瘤的TNM分期,风险度exp(β)为1.521,而Med1/TRAP220为预后有利因素,风险度exp(β)为0.386(P＜0.01).结论:Med1/TRAP220和uPAR联合应用在非小细胞肺癌预后预测中有重要意义.     

Mina53在胃癌组织中的表达及临床意义

目的:探讨mina53基因在胃癌组织中的表达及其与临床分期的相关性.方法:分别应用RT-PCR和免疫组化技术,检测71例胃癌患者组织标本以及20例癌旁组织中mina53基因和蛋白的表达水平,并分析其与胃癌临床分期的相关性.结果:(1)RT-PCR结果显示20例癌旁组织中仅2例mina53基因呈弱阳性表达,阳性率为10.00％;71例胃癌组织标本中有43例mina53基因表达阳性,阳性率为60.56％ (P＜0.05);其中Ⅰ、Ⅱ、Ⅲ、Ⅳ期胃癌组织标本中的阳性比例分别为11/23、15/29、12/14及5/5(P＜0.05).(2)免疫组化结果显示20例癌旁组织中仅3例mina53蛋白呈弱阳性表达,阳性率为15.00％;71例胃癌组织标本中有35例mina53蛋白表达阳性,阳性率为49.30％(P＜0.05);其中Ⅰ、Ⅱ、Ⅲ、Ⅳ期胃癌组织标本中的阳性比例分别为7/23、13/29、11/14及4/5 (P＜0.05).结论:与癌旁组织相比,mina53在胃癌中的基因和蛋白水平存在高表达,并且随肿瘤分期的进展而表达量增高,检测mina53表达水平有助于胃癌早期诊断和预测肿瘤的侵袭程度.     

NGX6a Is Degraded through a Proteasome-dependent Pathway without Ubiquitination Mediated by Ezrin,a Cytoskeleton-Membrane Linker

Our previous study demonstrated that the NGX6b gene acts as a suppressor in the invasion and migration of nasopharyngeal carcinoma (NPC). Recently, we identified the novel isoform NGX6a, which is longer than NGX6b. In this study, we first found that NGX6a was degraded in NPC cells and that this degradation was mediated by ezrin, a linker between membrane proteins and the cytoskeleton. Specific siRNAs against ezrin increase the protein level of NGX6a in these cells. During degradation, NGX6a is not ubiquitinated but is degraded through a proteasome-dependent pathway. The distribution pattern of ezrin was negatively associated with NGX6a in an immunochemistry analysis of a nasopharyngeal carcinoma tissue microarray and fetus multiple organ tissues and Western blot analysis in nasopharyngeal and NPC cell lines, suggesting that ezrin and NGX6a are associated and are involved in the progression and invasion of NPC. By mapping the interacting binding sites, the seven-transmembrane domain of NGX6a was found to be the critical region for the degradation of NGX6a, and the amino terminus of ezrin is required for the induction of NGX6a degradation. The knockdown of ezrin or transfection of the NGX6a mutant CO, which has an EGF-like domain and a transmembrane 1 domain, resulted in no degradation, significantly reducing the ability of invasion and migration of NPC cells. This study provides a novel molecular mechanism for the low expression of NGX6a in NPC cells and an important molecular event in the process of invasion and metastasis of nasopharyngeal carcinoma cells.     

一种新型细胞微阵列的制作和应用

为了高通量地检测大量培养细胞中基因原位表达, 发明了一种制作细胞微阵列的新方法,成功地制作含20种细胞系共100个供体细胞石蜡混合物点阵的细胞微阵列.免疫组化检测P53, P21, PTEN、P16基因在细胞微阵列中的蛋白质表达.原位杂交检测BRD7、NGX6 基因在细胞微阵列中mRNA原位表达.建立了P53、P21、PTEN、P16蛋白和BRD7、NGX6 mRNA 在不同培养细胞中的原位表达谱.细胞微阵列为基因功能研究提供一种新的高通量工具.细胞微阵列可广泛用于DNA、RNA和蛋白质水平上的基因原位表达研究.细胞微阵列还可用于筛选药物作用靶标的研究.     

Identification of a New Seven-span Transmembrane Protein: NGX6a Is Downregulated in Nasopharyngeal Carcinoma and Is Associated With Tumor Metastasis

Nasopharyngeal carcinoma (NPC)-associated gene 6 (NGX6) is a novel candidate metastasis suppressor gene that can significantly decrease the growth, motility, and invasion of NPC cells. In this study, we generated a highly specific NGX6 polyclonal antibody and analyzed its distribution in the human fetus by Western blot and immunohistochemistry. The result of the Western blot showed the protein of NGX6 had two types of isoforms, isoform a (NGX6a) and isoform b (NGX6b). Isoform a is composed of 472 amino acids with a calculated molecular mass of 52 kDa, whereas isoform b is composed of 338 amino acids with a calculated molecular mass of 37 kDa. It is predicated that there is an epidermal growth factor domain in the N terminal of both a and b isoforms, and seven transmembrane domains in NGX6a, but only two transmembrane domains in NGX6b. The expression level of NGX6a was higher than that of NGX6b in human fetal tissue. Obvious high expression of NGX6a protein presents in the nervous system and epithelial tissues of the human fetus, but the NGX6b protein (37 kDa) is mainly expressed in the nervous system. We further analyzed the tissue microarray, which contained 154 NPC biopsies and 70 non-NPC biopsies, and found that NGX6a was significantly downregulated in NPC and associated with tumor metastasis. (J Histochem Cytochem 58:41–51, 2010)     

NGX6基因抑制高转移鼻咽癌细胞的侵袭运动能力

 NGX6 基因是我室利用定位候选克隆策略,在鼻咽癌的高频杂合性缺失区9p21-22克隆的新基因.前期研究结果提示,它与鼻咽癌细胞的侵袭转移密切相关.为了进一步阐明其作用的结构基础,本研究成功构建了含NGX6基因及突变体的pCMV-myc瞬时和pcDNA3.1-his-myc(-)B稳定表达载体.通过脂质体转染技术,构建了NGX6及突变体的稳定表达细胞系5-8F.用免疫荧光试验和免疫电镜观察了NGX6在5-8F细胞中的亚细胞定位主要位于胞膜、核膜以及胞浆中的膜质结构,缺失突变的功能域对其定位没有明显的影响.基质胶侵袭试验和划痕试验研究了NGX6及突变体对细胞运动的影响.NGX6能抑制高转移潜能的鼻咽癌细胞5-8F的运动和侵袭能力,缺失胞内区（CYTO）后NGX6不能抑制5-8F细胞的运动和侵袭,提示CYTO可能是其发挥作用的重要功能域.     

NGX6基因转染对鼻咽癌细胞基因表达谱的影响

鼻咽癌是我国南方的常见肿瘤 .NGX6是新近克隆的定位于 9p2 1 2 2 ,在鼻咽癌组织中表达下调的基因 .初步的研究结果显示 ,NGX6基因转染鼻咽癌细胞HNE1能够延缓其生长速度 ,使肿瘤细胞更多地停留在G0 G1期 .为探索NGX6基因在鼻咽癌发病机制中的作用 ,建立了高表达NGX6的鼻咽癌细胞系 .利用包含 14 0 0 0个基因的cDNA微阵列分析了NGX6基因转染对HNE1细胞基因表达谱的影响 ,发现NGX6基因的转染能够上调p2 9、APC7、NEU1、RNasek6、αE catenin和TFⅡEα等基因的表达 ;同时也下调properdinP因子、G0S2、BAZ2B、ZHX1,OS4和PBX3等基因的表达 .研究结果提示 ,NGX6基因对鼻咽癌细胞的生物学行为的影响可能与它对一些细胞周期调控因子和转录调控因子的影响相关 .作为一种高通量的分析技术 ,cDNA微阵列为新基因的功能研究提供了重要的线索     

miR‐346 promotes migration and invasion of nasopharyngeal carcinoma cells via targeting BRMS1

The aim of this study is to determine the expression and roles of miR‐346 in nasopharyngeal carcinoma (NPC). We showed that miR‐346 was upregulated in NPC tissues compared with adjacent non‐tumorous nasopharyngeal tissues. Inhibition of miR‐346 significantly attenuated the migration and invasion of NPC cells. Luciferase reporter assay showed that miR‐346 targeted the 3′‐untranslated region (3′‐UTR) of breast cancer metastasis suppressor 1 (BRMS1). Overexpression of miR‐346 suppressed the endogenous expression of BRMS1 in NPC cells. There was a significant negative correlation between miR‐346 and BRMS1 protein expression in NPC tissues (r = ?0.372, P = 0.008). Rescue experiments demonstrated that overexpression of BRMS1 lacking the 3′‐UTR impaired the invasiveness of NPC cells transfected with miR‐346 mimic. Taken together, miR‐346 shows the ability to promote the migration and invasion of nasopharyngeal cancer cells via targeting BRMS1 and represents a potential therapeutic target for NPC.     

The tumor supressor function of STGC3 and its reduced expression in nasopharyngeal carcinoma

STGC3 is a novel candidate tumor suppressor gene that was found to be associated with nasopharyngeal carcinoma (NPC) via the cDNA cloning and RACE processes. The biological function of the STGC3 protein and its expression level in nasopharyngeal carcinoma remain unknown. This study aimed to evaluate the STGC3 protein expression level in NPC and to investigate the inhibitory function of STGC3 as a candidate tumor suppressor gene. We assessed the expression of the STGC3 protein in NPC biopsies and normal control specimens via Western blot and immunohistochemical analysis. The expression of STGC3 as induced by doxycycline (Dox) via a tetracycline (Tet)-regulated system in human nasopharyngeal carcinoma cell line CNE2 was also established, and the effect of STGC3 restoration on the biological behavior of CNE2 was observed. A reduced level of STGC3 expression (0.978 ± 0.213 versus 0.324 ± 0.185, P < 0.05) was detected in NPC versus normal nasopharyngeal tissue by Western blot assay. Immunohistochemical assays for STGC3 detected positive staining in the nuclei and cytoplasm of epithelial cells, and the positive expression rate in NPC, 8 of 21 (38%), was lower than that in normal nasopharynx samples, 16 of 22 (72%). After STGC3 expression was restored, the growth capacity and clone formation potential of CNE2 cells in soft agar were significantly suppressed, and the cell percentage in G₀/G₁ phase increased, while the percentage of cells entering the S and G₂ phases decreased. This indicates that an abnormality in STGC3 expression is associated with nasopharyngeal carcinogenesis and that it may play an important role in controlling cell growth and regulating the cell cycle.     

程序性死亡配体-1在鼻咽癌组织中的表达及临床意义

目的:探讨程序性死亡配体-1(PD-L1)在鼻咽癌组织中的表达及临床意义。方法:选取2015年7月-2017年6月期间在我院接受治疗的鼻咽癌患者333例,收集其鼻咽癌组织作为观察组标本,另选取同期在我院接受治疗的慢性鼻咽炎患者102例的鼻咽炎组织作为对照组标本。采用免疫组化法和逆转录-聚合酶链反应(RT-PCR)检测两组患者鼻咽组织中PD-L1蛋白和PD-L1 mRNA的表达,并分析观察组患者鼻咽癌组织中PD-L1蛋白和PD-L1 mRNA的表达与临床病理参数的关系。结果:对照组患者鼻咽炎组织中PD-L1蛋白的阳性率为0.00%(0/102),观察组患者鼻咽癌组织中PD-L1蛋白的阳性率为69.37%(231/333),两组PD-L1蛋白的阳性率比较差异有统计学意义(P0.05)。RT-PCR结果显示,对照组患者鼻咽炎组织中未见PD-L1 mRNA表达,观察组患者鼻咽癌组织中PD-L1 mRNA的相对表达水平为(0.82±0.27),差异有统计学意义(P0.05)。观察组患者鼻咽癌组织中PD-L1蛋白和PD-L1 mRNA的表达与年龄、性别无关(P0.05),而TNM分期为Ⅲ-Ⅳ期、有淋巴结转移、有吸烟史患者鼻咽癌组织中PD-L1蛋白阳性率和PD-L1 mRNA的表达均高于TNM分期Ⅰ-Ⅱ期、无淋巴结转移、无吸烟史患者(P0.05)。结论:在鼻咽癌组织中PD-L1蛋白和PD-L1 mRNA呈现高表达,且其表达与TNM分期、淋巴结转移、吸烟史有关。     

Expression of LINC00312, a long intergenic non-coding RNA, is negatively correlated with tumor size but positively correlated with lymph node metastasis in nasopharyngeal carcinoma

The long intergenic non-coding RNA LINC00312, also called NAG7, was first cloned by our group. Our previous studies have found that LINC00312 could inhibit proliferation and induce apoptosis in nasopharyngeal carcinoma (NPC) cells but also stimulate NPC cell invasion. However, the relevance of LINC00312 in NPC progression or in patient outcomes has not been reported. This study aims to assess the possible correlations of LINC00312 expression with NPC progression and its potential prognostic predictive ability in NPC outcomes. A NPC tissue microarray, which included 561 normal and NPC tissue cores, was used to detect LINC00312 expression, and we found that LINC00312 was significantly down-regulated in NPC tissues compared with non-cancerous nasopharyngeal epithelium tissues. Positive expression of LINC00312 was negatively correlated with tumor size (P < 0.001) but positively correlated with lymph node metastasis (P = 0.002). A receiver operating characteristic (ROC) analysis revealed that LINC00312 expression could distinguish non-cancerous patients from NPC patients (P < 0.001, sensitivity: 72.1 %, specificity: 87.7 %). We also found that LINC00312 was strongly negatively correlated with EBER-1, a non-coding RNA transcribed by Epstein-Barr Virus, in NPC (r = ?0.384, P < 0.001). In the final logistic regression analysis model, the abnormal expression of LINC00312 and EBER-1 were found to be independent contributors to nasopharyngeal carcinogenesis (P < 0.001, P < 0.001, respectively). A survival analysis revealed that LINC00312 could predict a good prognosis of no lymph node metastasis (Disease Free Survival (DFS): P = 0.005, Overall Survival (OS): P = 0.001) and a poor prognosis of lymph node metastasis (DFS: P = 0.011, OS: P = 0.001) in NPC patients. Low expression of LINC00312 was an independent risk factor for OS in multivariate analyses (P = 0.017). These observations indicated that LINC00312 could represent a potential biomarker for metastasis, progression and prognosis in NPC.