ERK/MAPK信号通路活性表达与鼻咽癌细胞增殖凋亡的关系

目的:探讨鼻咽癌细胞中ERK/MAPK信号传导通路异常激活与细胞增殖凋亡的关系.方法:SP法检测36例鼻咽癌、42例对照鼻咽黏膜上皮中p-ERK,Cyelin D1 和Ki-67蛋白的表达.用不同浓度阻滞刺PD98059处理CNE-2Z细胞,Hoechst33342/PI荧光双染法观察细胞凋亡,流式细胞仪检测凋亡,蛋白质印迹法检测Cyclin D1蛋白表达,免疫细胞化学法检测tyERK、Cyclin D1 和Ki-67蛋白的表达.结果:鼻咽癌组织中p-ERK、Cyclin D1和Ki-67的阳性表达率分别为69.44%(25/36)、80.56%(29/36)和80.56%(29/36),鼻咽黏膜上皮组织中则分别为23.80%(10/42)、19.05%(8/42)和9.52%(4/42),P<0.01.鼻咽癌组织中p-ERK-与Cyelin D1、Ki-67蛋白的表达呈正相关,r分别为0.604和0.668,P均<0.01.不同浓度PD98059作用于CNE-2Z细胞后,可观察到凋亡细胞,检测到凋亡峰,Cyclin D1蛋白表达的减少呈剂量依赖性,p-ERK、Cyclin D1和Ki-67蛋白的阳性表达细胞数减少.结论:鼻咽癌组织及CNE-2Z细胞中存在p-ERK蛋白的高表达和活化异常,p-ERK可能通过上调Cyclin D1和Ki-67的表达促进鼻咽癌细胞增殖并抑制凋亡.ERK信号通路有可能成为鼻咽癌治疗的新靶点.     

Cyclin D1、Ki-67在宫颈鳞状细胞癌中的表达及临床意义

目的 探索细胞周期蛋白Cyclin D1和细胞增殖活性标记物Ki-67在宫颈鳞状细胞癌中的表达及其与临床病理特征的相关性.方法 采用免疫组织化学方法检测宫颈鳞状细胞癌中Cyclin D1和Ki-67的表达，并进行相关分析。结果 Cyclin D1在宫颈鳞状细胞癌组织中阳性率为46．7％（14／30）；Ki-67在宫颈鳞状细胞癌组织中阳性率为73．3％（22／30）。Cyclin D1表达与有无转移呈正相关（P〈0．05），与病理分级及临床分期无明显相关性（P〉0．05）。Ki-67表达与病理分级及临床分期呈正相关（P〈0．05），与有无转移无明显相关性（P〉0．05）。结论 Cyclin D1和Ki-67的过表达可能与宫颈鳞状细胞癌的高侵袭性和不良预后密切相关。     

TGF-β1及其信号转导通路分子在鼻咽癌组织芯片中的表达及意义

目的探讨转化生长因子TGF-β1及其信号转导通路分子在鼻咽癌中的表达情况及临床病理意义。方法利用组织芯片及免疫组织化学技术检测163例鼻咽癌和42例鼻咽黏膜慢性炎组织中TGF-β1、TGF-βRⅠ、TGF-βRⅡ和Smad4蛋白的表达,分析其与各临床病理因素的关系。结果（1）TGF-β1在鼻咽癌组织中的阳性表达率为52.8％(86／163) ,明显高于鼻咽慢性炎组织中的阳性表达率19.0%（8/42）(P＜0．01);TGF-βRⅠ在鼻咽慢性炎组织中阳性表达率为57.1％(24／42),与鼻咽癌组织中的阳性表达率66.3％(108／163)比较差异无统计学意义（P＞0．05）;TGF-βRⅡ在鼻咽癌组织中的阳性表达率为49.1％(80／163),明显低于慢性鼻咽炎组织的阳性表达率71.4％(30／42) (P＜0．01);Smad4蛋白表达于胞质和(或)胞核中,在鼻咽癌组织中的阳性率为63.8％(104／163),明显低于鼻咽慢性炎组织的阳性表达率［83.3％(35／42) ］(P＜0．05)。（2）TGF-β1与Smad4的表达与鼻咽癌的临床分期和5年生存率有一定相关性(P＜0．05);TGF-βRⅠ在非角化性分化型鳞状细胞癌中的阳性表达明显低于非角化性未分化型鳞状细胞癌的表达（P＜0．05)。（3）TGF-β1、TGF-βRⅡ和Smad4蛋白的表达与鼻咽癌患者的年龄、性别和组织学类型无关,TGF-β1、TGF-βRⅠ、TGF-βRⅡ和Smad4蛋白在鼻咽癌组织中的表达不存在相关性(P>0.05)。结论TGF-β1、TGF-βRⅠ、TGF-βRⅡ和Smad4在鼻咽癌的发生、发展中起一定的作用。     

62例鼻咽癌中Ki67和CD44v6表达及意义

[目的]研究鼻咽癌中Ki67和CD44v6的表达及其关系。[方法]采用免疫组化S-P法检测23例鼻咽慢性炎、62例鼻咽癌（38例伴有淋巴结转移）组织中Ki67和CD44v6的表达情况。[结果]鼻咽慢性炎和鼻咽癌中Ki67阳性表达率分别为71.0%、13.0%,差异有统计学意义（P〈0.05）。CD44v6在鼻咽癌和鼻咽慢性炎中的阳性率分别为77.4%、17.4%,差异有统计学意义（P〈0.05）。鼻咽癌转移组Ki67和CD44v6的阳性率分别为84.2%、89.5%,明显高于非转移组的50.0%、58.3%（P〈0.05）。鼻咽癌中Ki67和CD44v6表达呈正相关（r=0.452,P〈0.05）。[结论]Ki67和CD44v6在鼻咽癌中的高表达协同参与了鼻咽癌的增殖和转移。     

Ki-67表达与鼻咽癌放射敏感性的关系

目的探讨Ki-67表达与鼻咽癌放射敏感性的关系。方法应用免疫组织化学法检测159例鼻咽癌组织中Ki-67蛋白表达的情况。结果159例鼻咽癌组织中Ki-67阳性表达率为98.7%。Ki-67表达强度与鼻咽癌的临床分期、根治放疗70Gy后鼻咽部肿瘤大小和颈部淋巴结的消退情况无关(P>0.05)。结论Ki-67与鼻咽癌的放射敏感性无关。     

MCM7和Ki-67在乳腺癌中的表达及其与新辅助化疗关系的研究

目的探讨乳腺癌新辅助化疗前后MCM7和Ki-67蛋白的表达状况,分析其与化疗疗效的关系。方法采用免疫组化方法检测55例乳腺癌新辅助化疗前后标本中MCM7和Ki-67的表达。结果新辅助化疗有效率为76.4%。化疗前MCM7和Ki-67蛋白阳性表达均显著高于化疗后（P〈0.01）;化疗前MCM7蛋白阳性表达显著高于Ki-67（P〈0.01）。化疗有效组（42例）MCM7蛋白阳性表达显著高于无效组（13例）（P〈0.01）,而Ki-67蛋白表达差异无统计学意义（P〉0.05）。化疗前MCM7、Ki-67表达呈正相关（r=0.58,P〈0.01）。结论ET方案新辅助化疗有较好的疗效,可能通过抑制MCM7、Ki-67蛋白的表达阻止乳腺癌细胞的增殖。MCM7蛋白高表达者化疗更敏感,MCM7可作为临床指导乳腺癌化疗并预测化疗敏感性的分子生物学指标之一。     

应用组织芯片技术研究人脑胶质瘤中β-Catenin、p53、Ki-67的表达及其相互关系

目的：探讨人脑胶质瘤中β-Catenin、p53的表达及与细胞增殖的相关性。方法：利用组织芯片技术及免疫组化方法检测β-Catenin、p53、Ki-67在正常脑组织、低/高级别星形细胞瘤中的表达,并分析其相关性。结果：正常脑组织中β-Catenin、p53、Ki-67表达均为阴性。β-Catenin、p53、Ki-67表达均随胶质瘤级别增高而增高,与组织分级密切相关（P〈0.01）,β-catenin在Ⅰ级胶质瘤中阳性率33.3%（2/6）,阳性部位为胞质;Ⅱ级阳性率28.6%（8/28）,阳性部位为胞质;Ⅲ级阳性率60.0%（18/30）,阳性部位为胞质,其中3例出现核阳性;Ⅳ级阳性率78.3%（18/23）,其中8例出现核阳性。β-Catenin与p53表达呈正相关（P〈0.01）。结论：β-Catenin的异常表达与胶质瘤的增殖活性增高有关,其与p53异常积聚可能相互协调促进胶质瘤的发生发展。应用组织芯片进行胶质瘤分子病理学研究较常规免疫组化方法有高通量、大样本以及经济快速等优势。     

p21WAF1、ETS-1、血管内皮生长因子-C蛋白在鼻咽癌组织中的表达及鼻咽癌内微淋巴管、微血管密度的临床病理意义

目的探讨细胞周期调控蛋白（p21^WAF1）、血管内皮生长因子-C（VEGF-C）及ETS-1蛋白在鼻咽癌（nasopharyngeal carcinoma,NPC）组织中的表达并以D2-40、CD34作标记测定微淋巴管密度（microlymphaticdensity,MLD）、微血管密度（microvessal density,MVD）,探讨它们之间的相关性及与鼻咽癌淋巴管及血管转移和预后的关系。方法应用免疫组化EnVision二步法及双标免疫组化染色,检测30例经治疗后生存时间〈5年的鼻咽非角化性癌（non-keratinizing carcinoma,NKC）,50例生存时间≥5年的NKC及对照组20例鼻咽黏膜慢性炎症组织（NP）内p21^WAF1、VEGF-C、ETS-1蛋白及MLD、MVD的表达水平。结果（1）p21^WAF1在NKC生存期〈5年组、生存期≥5年组和NP组中的阳性表达率依次递增,差异具有统计学意义（P〈0.05）。（2）VEGF-C、ETS-1蛋白表达及MLD、MVD在以上三组间的阳性表达率依次递减,差异有统计学意义（P〈0.05）。（3）本组NKC病例中p21^WAF1和VEGF-C经相关分析发现呈正相关关系,差异有统计学意义（P〈0.05）。（4）在80例鼻咽癌中VEGF-C与MLD、ETS-1与MVD呈正相关关系（P〈0.05）。结论检测鼻咽癌内p21^WAF1、VEGF-C、MLD、ETS-1及MVD可以作为预测NPC转移及判断预后的重要指标。     

ERK2和p-ERK1/2蛋白在宫颈癌中的表达和意义

目的：研究细胞外信号调节激酶2（extracellular regulated kinase 2,ERK2）及其磷酸化状态（p-ERK1/2）在宫颈癌的表达情况,探讨二者与宫颈癌侵袭、转移的关系。方法：采用免疫组化（SP）法,检测80例宫颈癌组织、50例CIN组织及30例正常宫颈组织中ERK2和p-ERK1/2的表达。结果：ERK2和p-ERK1/2在宫颈癌中阳性表达率为75%和52.5%,明显高于CIN组的3 4%和2 8%,以及正常宫颈组的13.33%和20%（P〈0.01）;ERK2和p-ERK1/2阳性表达率随临床分期的增高而增加（P〈0.05）,病理分级的降低而增加（P〈0.01）,有淋巴结转移者ERK2和p-ERK1/2阳性表达率明显高于无转移者（P〈0.01）;ERK2及p-ERK1/2在宫颈癌中的表达呈正相关（r=0.61,P〈0.01）。结论：ERK2及p-ERK1/2的表达与宫颈癌的生长、浸润、转移等生物学行为关系密切,在宫颈癌的发生发展过程中发挥重要作用。     

EZH2、Ki-67在成釉细胞瘤中的表达及其临床意义

目的 研究EZH2、Ki-67在成釉细胞瘤与正常口腔黏膜中的表达及两者之间的关系.方法 采用免疫组织化学SP法检测EZH2、Ki-67在50例成釉细胞瘤（30例原发、20例复发）和20例正常口腔黏膜中的表达.结果 EZH2在复发性成釉细胞瘤中蛋白阳性表达率为（36.25±7.24）％,高于原发性成釉细胞瘤的（25.26±4.28）％（P＜ 0.001）.Ki-67在复发性成釉细胞瘤中蛋白阳性表达率为（34.96±5.28）％,高于原发性成釉细胞瘤的（29.68±3.27）％（P＜0.05）.EZH2和Ki-67两者的表达呈正相关（P＜0.05）,但EZH2和Ki-67的表达与肿瘤的大小及患者性别无关（P＞0.05）.结论 成釉细胞瘤中存在EZH2和Ki-67的高表达,在成釉细胞瘤的发生、发展中起关键作用,与其复发有关,并可作为判定其预后的参考指标.联合检测EZH2和Ki-67蛋白的表达有助于为成釉细胞瘤的进程、复发的可能及其预后判断提供临床依据.     

Upregulated long non-coding RNA AFAP1-AS1 expression is associated with progression and poor prognosis of nasopharyngeal carcinoma

Altered expression of long noncoding RNAs (lncRNAs) associated with human carcinogenesis. We performed a cDNA microarray analysis of lncRNA expression in 12 cases of nasopharyngeal carcinoma (NPC) and 4 non-tumor nasopharyngeal epitheliums. One lncRNA, actin filament associated protein 1 antisense RNA1 (AFAP1-AS1), was identified and selected for further study. AFAP1-AS1 expression was upregulated in NPC and associated with NPC metastasis and poor prognosis. In vitro experiments demonstrated that AFAP1-AS1 knockdown significantly inhibited the NPC cell migration and invasive capability. AFAP1-AS1 knockdown also increased AFAP1 protein expression. Proteomic and bioinformatics analyses suggested that AFAP1-AS1 affected the expression of several small GTPase family members and molecules in the actin cytokeratin signaling pathway. AFAP1-AS1 promoted cancer cell metastasis via regulation of actin filament integrity. AFAP1-AS1 might be a potential novel marker that can predict cancer patient prognosis and as a potential therapeutic target for NPC.     

Insulin-like growth factor-1 content and pattern of expression correlates with histopathologic grade in diffusely infiltrating astrocytomas

Studies of experimental tumorigenesis have strongly implicated signaling of the insulin-like growth factor 1 (IGF-1) as a key component in astrocytic neoplasia; however, its role in the growth of low-grade and malignant human tumors is not well understood. Correlative analyses of IGF-1, p53, and Ki-67 (MIB-1) immunohistochemistry and IGF-1 receptor (IGF-1R) mRNA expression were performed to examine the cellular pattern of IGF-1 signaling in 39 cases of astrocytoma (World Health Organization grades II-IV). Tumor cells expressing IGF-1 and IGF-1R were present in all tumor grades. The proportion of tumor cells that expressed IGF-1 correlated with both histopathologic grade and Ki-67 labeling indices, while expression of IGF-1R mRNA correlated with Ki-67 indices. In cases where stereotactic tissue sampling could be identified with a specific tumor area by neuroimaging features, the numbers of IGF-1 immunoreactive cells correlated with the tumor zones of highest cellularity and Ki-67 labeling. In glioblastomas, the localization of IGF-1 immunoreactivity was notable for several features: frequent accentuation in the perivascular tumor cells surrounding microvascular hyperplasia; increased levels in reactive astrocytes at the margins of tumor infiltration; and selective expression in microvascular cells exhibiting endothelial/pericytic hyperplasia. IGF-1R expression was particularly prominent in tumor cells adjacent to both microvascular hyperplasia and palisading necrosis. These data suggest that IGF-1 signaling occurs early in astroglial tumorigenesis in the setting of cell proliferation. The distinctive correlative patterns of IGF-1 and IGF-1R expression in glioblastomas also suggest that IGF-1 signaling has an association with the development of malignant phenotypes related to aberrant angiogenesis and invasive tumor interactions with reactive brain.     

鼻咽癌发生发展过程中EBER1表达的研究

目的：探讨ＥＢＶ与鼻咽癌发生发展的关系及其病理学意义。方法：应用原位杂交技术检测５０例鼻咽癌石蜡切片组织中ＥＢＥＲ１的表达及分布状况。结果：正常鼻咽粘膜上皮及其单纯性增生、鳞状化生上皮ＥＢＥＲ１表达均阴性,鼻咽癌组织中表达率为９８％（４９／５０）,且在转移灶中不丢失。鼻咽异型增生上皮亦可表达ＥＢＥＲ１阳性信号（１６／２０）,结论：ＥＢＶ与鼻咽癌关系密切,ＥＢＶ可能在鼻咽上皮的恶性转化过程中起重要作用,鼻咽上皮的异型性改变尤其是ＥＢＥＲ１阳性者可认为是癌前病变,随访监测可能有利于早期发现鼻咽癌。     

Apaf-1和Ki-67在乳腺癌组织中的表达及临床意义

目的 探讨凋亡酶激活因子（Apaf-1）和Ki-67在乳腺癌组织中的表达并分析两者表达的临床意义。方法 收集2014年1月至2015年12月60例乳腺癌组织和30例癌旁组织存档蜡块,采用免疫组织化学 EnVision法检测Apaf-1和Ki-67的表达情况,同时分析两者表达的相关性及与临床病理特征（年龄、肿瘤大小、淋巴结转移、临床分期和身体质量指数）的关系。 结果 Apaf-1在乳腺癌组织中的阳性表达率为21.7%（13/60）,低于癌旁组织的70.0%（21/30）,差异有统计学意义（P<0.05）。Ki-67在乳腺癌组织中的阳性表达率为73.3%（44/60）,高于癌旁组织的36.7%（11/30）,差异有统计学意义（P<0.05）。两者表达与年龄、身体质量指数均无关（P>0.05）,与肿瘤大小、淋巴结转移及临床分期有关（P<0.05）。两者在乳腺癌组织中的表达呈负相关（r=-0.413,P=0.018）。结论 Apaf-1在乳腺癌中表达降低,而Ki-67水平升高,两者在乳腺癌的发展和侵袭中发挥促癌作用,有可能作为临床诊断乳腺癌的标志物。     

鼻咽癌变过程中DNA依赖蛋白激酶表达水平研究

Objective:To observe the expression of DNA-dependent protein kinase (DNA-PKcs) in the nasopharyngeal tissues during carcinogenesis.Methods:Patients including 30 cases of nasopharyngitis,42 cases of nasopharyngeal atypical hyperplasia and 34 cases of nasopharyngeal carcinoma (NPC) were chosen from Zhongshan area in which there is high incidence rate of NPC.The expression of DNA-PKcs in the nasopharyngeal tissue was examined by immunohistochemistry.Results:The expression rates of DNA-PKcs in the nasopharyngitis tissue,the nasopharyngeal precancerous lesion and the NPC were 6.7%,64.3% and 55.9%,respectively (P＜0.001).Conclusion:The expression of DNA-dependent protein kinase may play an important role in nasopharyngeal tissue carcinogenesis.     

细胞周期素D1、Ki67在鼻咽癌中的表达及意义

This study was conducted to detect the expression of cyclinD1 and Ki67 in nasopharyngeal carcinoma (NPC) and their correlation with the biological and prognosis. 56 cases of biopsy specimens of NPC which had been embedded with paraffin in 1996 in our hospital were collected and immunostained with cyclinD1 and Ki67 monoclonal antibodies by means of the streptavidin peroxides method. The patients were followed up periodically, and then their biological behaviors and prognosis were statistically analyzed. The percentage of cyclinD1 and Ki67 positive cells in the NPC specimens ranged from 0–54% and 0–31% respectively. The staining was nuclear. Of the 56 cases, 30 cases (56.6%) highly expressed cyclinD1 or Ki67 HPI and 26 cases (46.4%) lowly expressed cyclinD1, while only 16cases (28.6%) showed Ki67 HPI and 40 cases (71.4%) showed Ki67 LPI. Patients who lowly expressed cyclinD1 or highly expressed Ki67 had a higher radiosensitivity and a better prognosis. Based on these findings, cyclinD1 and Ki67 immunohistochemical staining is considered to be useful, not only as an independent factor of radiosensitivity and prognosis respectively, but also as a means of determining the optimum treatment for each individual patient NPC.     

鼻咽癌组织中EGFR和p-ERK蛋白表达的检测及意义

目的 检测鼻咽癌组织中EGFR信号传导通路相关分子EGFR、p-ERK的表达,并对其与鼻咽癌患者预后的关系进行探讨。方法 采用免疫组化LSAB法对鼻咽癌组织中EGFR、p-ERK的表达进行检测,用统计学软件SPSS 10．0进行数据处理,分析蛋白表达与鼻咽癌的临床病理特征及预后的关系。结果 EGFR阳性表达率为70．9％;EGFR蛋白的表达与鼻咽癌临床分期（1992年全国第六届鼻咽癌会议制定）及性别相关。EGFR阳性患者的总生存期（OS）、疾病进展时间（TTP）显著低于阴性患者。p-ERK阳性表达率为52．7％;临床分期Ⅰ、Ⅱ期患者p-ERK蛋白的表达率明显低于Ⅲ、Ⅳ期患者;p-ERK阳性患者的OS、无病生存率（DFS）、TTP皆显著低于阴性患者。鼻咽癌组织中EGFR表达与p-ERK表达呈正相关。Cox多因素分析显示,年龄〉50岁是独立的不良预后因素,而EGFR和p-ERK的表达均不是独立的预后因素。结论 鼻咽癌组织中存在EGFR、p-ERK高表达。EGFR阳性患者的0S、TTP显著低于阴性患者;p-ERK阳性患者的OS、DFS、TTP均显著低于阴性患者。EGFR和p-ERK的表达均不是鼻咽癌独立的预后因素。     

Interferon-induced protein 16 expression in colorectal cancer and its correlation with proliferation and immune signature markers

Interferon-induced protein 16 (IFI16) is important for innate immune recognition of foreign/damaged DNA. Abnormal IFI16 expression is closely related to the occurrence of multiple malignant tumours, but its expression pattern in colorectal cancer (CRC) remains unclear. The present study aimed to investigated IFI16 expression and association with cell proliferation in CRC tissues and adjacent normal tissues. A multiplex immunofluorescence panel of antibodies against IFI16, Ki-67 and phosphorylated (p)-ERK1/2 was applied to assess a tissue microarray (TMA). The TMA included 77 CRC samples and 74 normal adjacent tissue samples which were collected from The First People''s Hospital of Yunnan Province (Kunming, China) (3 paracancerous tissues were lost because of repeated cutting). Immunohistochemistry was used to detect CD8⁺ tumour-infiltrating lymphocyte (TIL) abundance and programmed death-ligand 1 (PD-L1) expression in cancer tissues. The present study demonstrated that IFI16 localized to the nucleus of CRC cells. Although IFI16 was weakly expressed in normal mucosal epithelial cells, absent to strong expression was detectable in different patients with CRC. Typically, IFI16 was not co-localized with Ki-67 within CRC cells. The multiplex immunofluorescence data demonstrated that the proportion of IFI16⁻/Ki-67⁺ cells from CRC tissues was 57.13%; however, that of IFI16⁺/Ki-67⁺ cells was 1.50%. The IFI16⁻/Ki-67⁺ phenotype was significantly positively associated with the tumor-node-metastasis stage and was marginally significantly correlated with lymph node metastasis. p-ERK1/2 protein was primarily localized to the cytoplasm and cell membrane of CRC cells and sometimes to the nucleus. Although, IFI16 demonstrated a strong correlation with p-ERK1/2, IFI16 did not co-localize with p-ERK1/2 and the proportion of IFI16 and p-ERK1/2 double-negative CRC cells was 84.95%. IFI16 expression displayed no significant association with CD8⁺ TILs or PD-L1. However, a strong positive correlation between CD8⁺ TILs and PD-L1 was observed. High CD8⁺ TIL infiltration in CRC tissue was associated with lower lymph node metastasis and tumor-node-metastasis stage. In summary, the results of the present study provided a novel insight for the role of IFI16 in CRC occurrence via the regulation of cancer cell proliferation.