细胞周期调节蛋白在食管鳞癌组织中的表达

背景与目的:研究表明肿瘤细胞周期分析显示细胞具高增殖率的肿瘤,其临床病情发展快;细胞周期调节蛋白Ki-67、细胞周期蛋白A及p27介入到细胞的增殖,但这些因子与食管癌之间的关系的研究在我国尚未见报道。本研究观察细胞周期调节蛋白Ki-67、细胞周期蛋白A及p27在中国食管癌患者中的表达特征,以探讨这些分子标志物与临床病理因素之间的关系。方法:60例(48例男性、12例女性)食管鳞癌患者行外科手术切除肿瘤,其标本行免疫组织化学染色。Ki-67及细胞周期蛋白A表达程度以染色指数表示,p27以标记指数表示。结果:Ki-67、细胞周期蛋白A及p27免疫组化染色在瘤组织与非瘤组织中均固定于细胞核。Ki-67及细胞周期蛋白A的染色指数在低分化鳞癌(27.2±4.9;15.4±5.3)明显高于高分化鳞癌(20.6±6.3;11.3±6.4,P<0.05);p27免疫组化染色的阳性率在高分化鳞癌(36%)高于其它病理类型(29%及18%),但相互之间的差异无统计学意义(P>0.05)。结论:Ki-67、细胞周期蛋白A及p27的表达程度可反映食管鳞癌细胞的增殖状况。细胞周期调节蛋白Ki-67及细胞周期蛋白A的过度表达提示食管鳞癌细胞分化差。     

食管鳞癌p16和p53蛋白的表达及其意义

目的 :探讨 p16、p5 3蛋白在食管鳞癌 (ESC)中的表达及其意义。方法 :利用 S- P法检测 5 6例 ESC中 p16和 p5 3蛋白的表达。结果 :5 6例食管鳞癌中 ,p16蛋白表达阳性 2 1例 ,占 37.5 % ,p5 3蛋白表达阳性 35例 ,占 6 2 .5 % ,p16和 p5 3蛋白表达与肿瘤分化程度关系密切 ,随分化程度的降低 ,p16蛋白阳性率逐渐降低 (P<0 .0 2 5 ) ,p5 3蛋白阳性率逐渐增加 (P<0 .0 5 ) ,p16阳性表达组 p5 3表达显著低于 p16阴性组 (P<0 .0 1) ,且与淋巴结转移有一定的关系。结论 :联合检测食管鳞癌组织中 p16和 p5 3蛋白的表达有助于综合判断食管鳞癌的恶性程度和转移潜能。     

p53、bcl-2蛋白和p-gp在鼻咽鳞癌中表达的研究

 目的　研究 p5 3、bcl 2蛋白和 p gp在鼻咽鳞癌组织中的表达及其临床病理特征的关系 ,探讨其在预后评估中的意义。方法　采用免疫组化SP法检测 6 4例鼻咽鳞癌组织中p5 3、bcl 2和 p gp的表达。结果　鼻咽鳞癌组织中 p5 3、bcl 2和p gp阳性率分别为76 .5 6 % (4 9/ 6 4 )、89.0 6 % (5 7/ 6 4 )、17.19% (11/6 4 )。p5 3、bcl 2蛋白过表达均与颈部淋巴结转移、组织分化程度有关 (P <0 .0 1)。p gp过表达与bcl 2蛋白过表达呈正相关 (P <0 .0 5 ) ,p5 3蛋白过表达与bcl 2蛋白过表达呈正相关 (P <0 .0 0 1)。结论　p5 3和bcl 2蛋白均参与鼻咽鳞癌的发生与分化 ,其表达水平的不同有助于预后判断 ;未经治疗的鼻咽鳞癌存在原发性耐药     

p27和cyclin E在胆囊癌中表达及与临床病理关系的研究

目的 :探讨p2 7蛋白及细胞周期素E(cyclinE)与胆囊癌发生和发展的关系。方法 :应用免疫组化SABC法检测 5 4例胆囊癌、4 8例胆囊腺瘤及 5 0例胆囊正常黏膜中p2 7及cyclinE表达情况 ,同时结合临床病理资料进行分析。结果 :5 4例胆囊癌中 ,8例 (15 % )p2 7高表达 ,4 4例 (81 4 8% )cyclinE表达 ;4 8例胆囊腺瘤中 ,2 9例 (6 0 % )p2 7高表达 ,2 7例 (5 6 2 5 % )cyclinE表达 ;5 0例胆囊正常黏膜中 ,35例 (70 % )p2 7高表达 ,2 6例 (5 2 % )cyclinE表达。p2 7蛋白在胆囊癌中的表达明显降低 ,三者之间差异有极显著意义 ,P <0 0 0 5。另外 ,p2 7在胆囊癌中表达与癌细胞的分化程度差异有显著意义 ,P <0 0 5 ;低分化癌p2 7表达较低 ,高分化与低分化的p2 7表达差异有显著意义 ,P <0 0 5。p2 7低表达的患者 1年生存率显著降低。cyclinE在胆囊癌中表达与癌细胞的分化程度和肿瘤的病理分期有关。p2 7和cyclinE在胆囊癌中表达存在着相反的联系。结论 :p2 7是细胞周期的负调控因子和潜在的肿瘤抑制因素。p2 7蛋白的降低和cyclinE的过表达在胆囊癌的发生中可能起着重要作用。p2 7是胆囊癌预后的一个可靠指标。     

P16蛋白表达与原发性食管鳞癌患者临床病理相关性研究

目的　探讨抑癌基因p16在食管鳞癌组织中的蛋白表达与临床、病理变化之间的关系。方法　应用改良SABC免疫组化法 ,对 5 4例手术切除的原发食道鳞癌患者的肿瘤组织蜡块进行p16基因蛋白表达产物的测定。结果　 5 4例食管鳞癌病人中p16基因表达缺失 2 1例 ( 38.9% )。p16表达与肿瘤病理分级关系密切 ,p16表达缺失者肿瘤组织分化程度明显低于p16表达阳性者 (P <0 .0 0 0 1)。p16基因表达与区域淋巴结转移也有一定的相关性。结论　原发性食管鳞癌患者的p16基因蛋白表达缺失较常见 ,且与肿瘤组织的分化程度密切相关 ,与淋巴结转移也有一定的关系。利用SABC免疫组化法检则p16基因的表达情况 ,可作为原发性食管鳞癌的辅助诊断手段 ,同时也为选择恰当的治疗方案提供了重要的信息。     

细胞周期蛋白D1和E在食管鳞状细胞癌中的表达及意义

目的　探讨细胞周期蛋白 D1 和 E( cyclin D1 and cyclin E)在食管鳞状上皮癌中的表达及意义。方法　采用 S- P免疫组化方法 ,检测了周期蛋白 D1 和 E在正常食管上皮 ( 2 0例 )、增生上皮 ( 2 1例 )和鳞癌 ( 6 2例 )中的阳性率。结果　正常鳞状上皮中 cyclin E无阳性表达 ,在增生上皮中 cyclin E阳性率为 33.3% ,在鳞癌中阳性率为5 3.2 % ;cyclin D1 在正常鳞状上皮阳性率为 5 .0 % ,在增生和鳞癌中阳性率分别为 4 2 .9%和 4 6 .8% ;cyclin E在低分化鳞癌组阳性率 ( 80 .0 % )明显高于高分化组 ( 2 9.0 % ) ( P<0 .0 5 )。 cyclin D1 和 cyclin E在鳞癌中的阳性表达呈正相关 ( r=0 .782 )。结论　 cyclin D1 和 cyclin E过表达与食管鳞状细胞癌的发生和分化程度有关。     

食管鳞癌与p53及CD44v6表达分析

目的　分析 p5 3、CD4 4v6与食管鳞癌的预后关系。 方法　用免疫组化法测定原发食管鳞癌 p5 3癌基因、CD4 4v6的表达。结果　p5 3的阳性表达为 4 7.8% ,与肿瘤的分化程度、浸润深度呈显著相关 (P <0 .0 5 ) ,与淋巴结有无癌转移及肿瘤大小未见显著相关 (P >0 .0 5 )。CD4 4v6阳性表达为 79.9% ,分化程度高的阳性表达大于分化程度低的 ,侵及外膜、淋巴结有癌转移的表达大于无外膜受侵和无淋巴转移 ,但未见显著性 (P >0 .0 5 )。结论　p5 3阳性表达预示食管鳞癌分化程度低 ,有较强的侵袭能力。CD4 4v6作为对食管癌的浸润转移和预后标记物并不十分理想。     

食管鳞癌中PTEN蛋白的表达及意义

目的　通过对PTEN在食管鳞癌中表达的研究,探讨其与食管鳞癌的发生、浸润、转移的关系。方法　应用免疫组织化学S P法检测49例食管鳞癌组织及40例正常食管黏膜组织中PTEN的表达。结果　食管鳞癌组织中PTEN蛋白表达率为5 3 0 6% (2 6/4 9)。显著低于正常食管黏膜组织的表达10 0 % (4 0 /4 0 ) ,在不同分化程度的癌组织中其阳性表达率有显著性差异(P <0 0 5 ) ,且PTEN蛋白表达率随癌组织浸润深度的加深而明显降低(P <0 0 5 ) ,有淋巴结转移组明显低于无淋巴结转移组(P <0 0 5 )。结论　PTEN基因缺失或蛋白表达降低在食管鳞癌的发生发展中可能起重要作用,且与肿瘤的分化程度、浸润转移密切相关。     

原发性食管鳞癌组织p16基因缺失的检测

目的　研究p16基因缺失与原发性食管鳞癌的相关性。方法　采用PCR方法检测 41例原发性食管鳞癌组织中p16基因的缺失情况。结果　 41例食管鳞癌组织有 6例p16基因缺失 ,缺失率为 14 .6%。低分化食管癌p16基因缺失率为 5 0 % ,高于中分化 (p =0 .0 0 0 9)和高分化食管癌 (p =0 .0 2 2 )。有淋巴结转移缺失率高于无淋巴结转移 (p =0 .0 0 3 )。结论　p16基因缺失与食管鳞癌的发生发展有相关性。     

食管黏膜上皮癌变过程中p27、cyclinD1和bcl-2蛋白的表达及其意义

目的　探讨p2 7、cyclinD1和bcl 2基因在食管鳞癌发生发展中的意义和相互间的作用。方法　采用免疫组化染色检测p2 7、cy clinD1和bcl 2蛋白在正常食管黏膜、非典型增生及浸润癌各 64例中的表达。结果　p2 7在正常食管黏膜表达率最高 (90 6% ) ,随食管黏膜上皮病变的发展表达率呈下降趋势 ,而cyclinD1和bcl 2在食管鳞癌的发生发展中表达呈上升趋势 ,p2 7、cyclinD1和bcl 2的表达率在非典型增生组、浸润癌组与正常组比较均有显著性差异 (P <0 0 1) ;相关分析显示食管鳞癌组织中p2 7与cyclinD1呈负向表达 ;p2 7、cyclinD1和bcl 2的表达与食管癌的淋巴结转移密切相关 (P <0 0 5 )。结论　食管癌的发生发展是多因素作用的结果 ,p2 7、cyclinD1和bcl 2的表达与食管癌的临床病理指标有关 ,可作为食管鳞癌早期诊断和判断预后的重要分子指标。     

食管癌组织上皮细胞黏附分子表达的临床意义

 【摘要】　目的　探讨上皮细胞黏附分子（Ep-CAM）在食管鳞状细胞癌组织中表达的临床意义。方法　应用免疫组织化学方法检测70例食管正常黏膜、癌组织和72枚区域淋巴结内Ep-CAM的表达情况。结果　在正常食管组织中，Ep-CAM未见阳性表达；但在食管癌组织中表达阳性率为94.3 %；其表达强度与食管癌的病变长度、浸润深度无明显相关，但与肿瘤分化程度、淋巴结是否转移明显相关（P＜0.001）；Ep-CAM表达强度与患者术后3年生存率呈明显负相关（P＜0.001）。结论　Ep-CAM在食管癌组织中特异性高表达，可作为食管癌诊断、预后判断及治疗的有用指标。     

Significant correlation between expression of heat shock proteins 27, 70 and lymphocyte infiltration in esophageal squamous cell carcinoma

The objective of this study was to clarify the clinicopathologic and prognostic significance of heat shock proteins (HSP) 27 and 70 expression in esophageal squamous cell carcinoma (SCC). Immunohistochemical staining for HSPs 27 and 70 was performed on surgical specimens obtained from 62 patients with esophageal SCC. The expression of both HSPs 27 and 70 correlated inversely with depth of invasion (P<0.05) and pathologic stage (P<0.05), and correlated positively with lymphocyte infiltration (P<0.05). Reduction of HSP 70 expression was significantly correlated with poor prognosis (P<0.05). Patients with HSP 27-negative tumors tended to have a poor prognosis compared with patients with HSP 27-positive tumors. The present findings suggest that HSPs 27 and 70 are significant prognostic factors for esophageal SCC.     

IL-27基因修饰树突状细胞活化的特异性CTL对人食管癌裸鼠移植瘤生长增殖的影响

研究IL-27基因转染树突状细胞（DC）体内诱导免疫杀伤食管癌细胞的效能及其机制。方法:基因转染的方法建立表达IL-27基因的树突状细胞（DC/IL-27）;构建人食管癌细胞裸鼠移植瘤模型,皮下注射食管癌细胞抗原致敏、IL-27基因修饰DC（DC/IL-27-Ag）活化的特异性CTL后观察荷瘤裸鼠抑瘤率;TUNEL法检测荷瘤裸鼠肿瘤细胞的原位凋亡;流式细胞术检测移植瘤细胞的细胞周期、凋亡率。结果:RT-PCR显示DC/IL-27细胞中有IL-27 p28和EBI3亚基基因表达提示转染成功;免疫接种DC/IL-27-Ag活化的CTL组的抑瘤率为58.28%,明显高于其他组,差异有统计学意义（P<0.01）。TUNEL法检测显示,DC/IL-27-Ag活化的CTL组凋亡率明显高于其他组（P<0.01）,流式细胞术（FCM）显示肿瘤组织内细胞增殖指数为（23.92±1.60）%,显著低于其他组,差异有统计学意义（P<0.01）;细胞凋亡率为（32.78±0.83）%,显著高于其他组,差异有统计学意义（P<0.01）。结论:DC/IL-27-Ag可活化特异性CTL,在裸鼠体内产生了抑制肿瘤生长的作用,显著抑制食管癌细胞增殖、促进其凋亡,为DC应用于食管癌的免疫治疗提供了理论和动物实验依据。      

Elevated focal adhesion kinase expression facilitates oral tumor cell invasion

BACKGROUND: Understanding the molecular mechanisms of metastasis is critical with respect to oral tumorigenesis. The focal adhesion kinase (FAK) is an intracellular tyrosine kinase associated with the regulation of cell growth, migration, and survival. The purpose of the current study was to determine whether elevated FAK expression in oral malignancies was associated with increased invasiveness and oral carcinoma. METHODS: Immunohistochemical analysis was used to assess levels of FAK expression in archived oral carcinoma tissue samples. Invasion assays after transfections were used to assess the effect of increased FAK expression on invasive potential of oral tumor cells. RESULTS: The human oral carcinoma cell line SCC25 was significantly more invasive (P < 0.05) and expressed higher levels of FAK compared with the less invasive human oral carcinoma cell line SCC15. FAK expression was 3.0-fold higher in the SCC15 cell line and 5.0-fold higher in the SCC25 cell line compared with normal epithelial cells. In the highly invasive SCC25 cell line, FAK expression was 1.5-fold higher compared with the less invasive SCC15 cell line. FAK immunostaining in oral tumors was significantly more intense compared with the immunostaining in surrounding normal epithelium or chronic mucositis. Overexpression of FAK in low-invading SCC15 cells resulted in a 4.5-fold increase in the rate of invasion compared with untransfected or neotransfected control SCC15 cell lines and a nearly 1.5-fold greater rate compared with the highly invasive untransfected SCC25 cell line. CONCLUSIONS: The current results suggest that enhanced expression of FAK in oral carcinoma cells may lead to a selective growth advantage and increased invasive potential of the primary oral tumor.     

Prolyl isomerase Pin1 expression predicts prognosis in patients with esophageal squamous cell carcinoma and correlates with cyclinD1 expression

Esophageal carcinoma is one of the most lethal tumors, and identification of prognostic factors for patients with this disease is important. Propyl isomerase Pin1 is overexpressed in some human cancers and thought to be an important regulator of cyclinD1. However, the relationships between Pin1 expression and clinicopathologic features in patients with esophageal squamous cell carcinoma (SCC) have not been explored. Here, we investigated the role of Pin1 in association with cyclinD1 in esophageal SCC progression and its clinicopathological significance. The expressions of Pin1 and cyclinD1 were examined immunohistochemically in surgical specimens from 119 esophageal SCC patients. The expression levels of Pin1 and cyclinD1 in 6 esophageal SCC-derived cell lines were compared with those in an immortalized human esophageal cell line by western blotting. Pin1 overexpression was correlated with lymph node metastasis (P=0.0384), and its expression was related to cyclinD1 expression. Pin1 expression was correlated with poor prognosis in esophageal SCC patients (P=0.0044), and found to be an independent prognostic factor (P=0.0277). Pin1 was overexpressed in 5 of 6 esophageal SCC-derived cell lines compared with immortalized esophageal keratinocytes. Moreover, the Pin1 level was correlated with the cyclinD1 level in 4 of the 6 cell lines. In conclusion, Pin1 expression is correlated with cyclinD1 expression and may be a useful prognostic factor for esophageal SCC.     

Tumor-suppressive effect of retinoid receptor-induced gene-1 (RRIG1) in esophageal cancer

We previously showed that induction of retinoid receptor-induced gene-1 (RRIG1) expression inhibited RhoA activation and tumor cell colony formation, invasion, and proliferation, and these effects are associated with the suppression of extracellular signal-regulated protein kinases 1 and 2 phosphorylation and cyclooxygenase-2 expression. To further elucidate its role in tumor cell growth, gene expression, and tumorigenesis, we determined RRIG1 expression in breast and esophageal tissue specimens and then stably transfected RRIG1 into a TE-8 esophageal squamous cell carcinoma (SCC) cell line. We found that RRIG1 was expressed in normal mammary glands (10 of 10) but not all ductal carcinoma in situ [11 of 19 (57.9%), P = 0.018] and invasive cancer [14 of 30 (46.7%), P = 0.0023] tissues. Similarly, RRIG1 was expressed in normal esophageal epithelium (22 of 22) but not all dysplastic [6 of 43 (14%), P = 0.0001] and SCC [50 of 122 (41%), P = 0.0001] tissues. Furthermore, RRIG1 expression correlated positively with tumor differentiation but inversely with lymph node metastasis of esophageal SCC. Finally, the stable transfection of RRIG1 inhibited esophageal SCC cell growth and the expression of extracellular signal-regulated protein kinases 1 and 2 and cell cycle-related genes (e.g., cyclin D1, phosphorylated Rb, and E2F). RRIG1-transfected sublines also inhibited tumor development in nude mice. The results of this study indicate that RRIG1 plays a role in suppressing tumorigenesis.     

Lack of activated Smad2 in transforming growth factor-beta signaling is an unfavorable prognostic factor in patients with esophageal squamous cell carcinoma

BACKGROUND AND OBJECTIVES: Transforming growth factor-beta (TGF-beta) regulates cell growth in various cells, and inactivation of the TGF-beta-signaling pathway contributes to tumor progression. In this study, we investigated the expression of Smad2 and Smad3, which are specific intracellular mediators of TGF-beta signaling. We also examined the relationship between the expression levels of activated Smad2 by TGF-beta and clinicopathologic characteristics of patients with esophageal squamous cell carcinoma (SCC). METHODS: Immunohistochemical staining with anti-phosphorylated Smad2 (P-Smad2) polyclonal antibody, anti-Smad2 monoclonal antibody, and anti-Smad3 polyclonal antibody was performed on surgical specimens obtained from 80 patients with esophageal SCC. RESULTS: Our data indicated that a low level of P-Smad2, as detected immunohistologically, correlated with lymph node metastasis (P = 0.0002), distant metastasis (P = 0.0338), pathologic stage (P = 0.0093), and poor survival rate (P = 0.0246). All patients without positive Smad2 immunostaining were included among those without positive P-Smad2 immunostaining. There was no significant correlation between expression of Smad2 or Smad3 and clinicopathologic characteristics. CONCLUSIONS: We demonstrated that a lack of Smad2-P appears to be correlated with tumor development and poor prognosis in patients with esophageal SCC.     

食管上皮癌变过程中CDK4、p18、p19的表达及意义

 【摘要】　目的　探讨细胞周期调控蛋白CDK4、p18、p19在食管鳞状细胞癌（SCC）发生、发展中的作用。方法　制作组织芯片,用免疫组织化学EnVision二步法对120例食管癌患者手术标本中CDK4、p18、p19的表达进行检测,并对其结果进行统计分析。结果　CDK4蛋白在正常食管上皮的表达低[28.3 %（34／120）],瘤变上皮中有所增高[32.5 %（39／120）],食管SCC中表达高[84.2 %（101／120）],且随SCC分化程度的降低而逐渐增高,SCC与正常上皮及瘤变上皮中CDK4表达阳性率差异有统计学意义（χ2＝76.004,P＜0.05;χ2＝65.897,P＜0.05）。淋巴结转移组CDK4表达率[93.88 %（46／49）]高于无淋巴结转移组[71.43 %（55／71）]（χ2＝5.860,P＜0.05）。p18、p19 蛋白在正常食管上皮组织阳性表达率分别为34.2 %（41／120）、29.2 %（35／120）,在食管瘤变上皮中表达率分别为19.2 %（23／120）、15.0 %（18／120）,在SCC中表达率分别为63.3 %（76／120）、61.7 %（74／120）,两指标在正常上皮及瘤变上皮间、瘤变上皮与SCC间、正常上皮与SCC间差异均有统计学意义（p18：χ2＝6.903、48.296、20.429,均P＜0.05;p19：χ2＝6.998、55.276、25.565,均P＜0.05）;在食管SCC中随分化程度的降低而逐渐增高。p18、p19分别与CDK4基因表达呈正相关（r＝0.696、0.630,均P＜0.05）。p18与p19二者呈正相关（r＝0.833,P＜0.05）。结论　细胞周期调控基因CDK4 、p18、p19参与食管SCC的发生、发展,其蛋白表达与食管上皮癌变密切相关。     

Serum concentration and expression of Reg IV in patients with esophageal cancer: Age-related elevation of serum Reg IV concentration

Regenerating islet-derived family, member 4 (REG4, which encodes Reg IV) is a marker for cancer and inflammatory bowel disease. This study aimed to investigate the diagnostic utility of Reg IV measurement in sera from esophageal cancer patients. Reg IV expression was examined in 269 esophageal cancer samples by immunostaining and the Reg IV levels in sera were measured from 65 patients with esophageal squamous cell carcinoma (SCC) by enzyme-linked immunosorbent assay. No Reg IV staining was detected in 255 SCC and 4 small cell carcinoma samples, whereas Reg IV was stained in 4 of 10 (40%) adenocarcinoma samples. Serum Reg IV concentration in esophageal SCC patients was significantly higher compared to that of the control subjects (P=0.0003). A significant correlation between serum Reg IV concentration and age was found in control subjects (P<0.0001). When serum Reg IV concentration was analyzed according to age, the distribution of serum Reg IV concentration in patients with esophageal SCC was similar to that of the control subjects. These results suggest that Reg IV expression is highly specific for adenocarcinoma of the esophagus. Further investigation is required to clarify whether Reg IV serves as a serum tumor marker for esophageal cancer.