CLINICAL SIGNIFICANCES OF THE EXPRESSION OF METALLOTHIONEIN IN FIVE TYPES EPITHELIUM CELL CANCER

Objective： To study the expressions of （CSC）, bladder transitional cell cancer metallothionein and the significances in cervical squamous cell cancer （BTC）, esophageal squamous cell cancer （ESC）, gastral tubular adenocarcinoma （GC） and large intestinal tubular adenocarcinoma （LIC）. Methods： lmmunohistochemical method was used to examine the expression rates of MT in five types of cancer tissue. Results： The expression rates of MT were 75.00% （24/32） in ESC, 52.27% （46/88） in GTC, 59.46% （44/74） in LIC, 64.86% （48/74） in BTC and 58.57% （41/70） in CSC respectively. The positive rates of MT expression were higher in low differentiation and deep muscular group than those in medium or high differentiation and superficial muscular invasion group （P〈0.05）. Conclusion： The expression of MT is related to differentiation degree and invasion degree.     

Relationship between Preoperative Clinicopathologic Characteristics and Lymph Node Metastasis in Early Gastric Cancer

Objective： To investigate the features of the preoperative clinicopathologic characteristics in correlation with lymph node metastasis. Methods： The preoperative clinicopathologic characteristics and lymph node metastasis of 265 patients with early gastric carcinoma were analyzed retrospectively. Results： The three clinicopathologic characteristics, maximum cancer diameter 〉2cm under endoscope, poor differentiation and excavated type were significant high risk independent preoperative clinicopathologic characteristics （P〈0.05） . The patients who had none of the three preoperative clinicopathologic characteristics had no lymph node metastasis, while 27.27% of the patients who had all the three preoperative clinicopathologic characteristics had N2 lymph node metastasis. Conclusion： The three preoperative clinicopathologic charaeteristics, maximum cancer diameter under endoscope, cell differentiation and gross type were very useful to evaluate the extent of lymph node metastasis.     

Expression and significance of the polar regulationassociated protein in cholangiocarcinoma

Objective： To investigate the expressions of atypical protein kinase C = subtype （aPKC-I and E-cadherin in cholangiocarcinoma, and analyze molecular mechanisms of the invasion and metastasis of cholangiocarcinoma. Methods： The expressions of aPKC-I nd E-cadherin in 9 specimens of benign bile duct tissues and 35 specimens of cholangiocarcinoma were detected by EnVision immunohistochemistry, and their correlations to the clinicopathologic characteristics and invasion of cholangiocarcinoma were analyzed. Results： The positive rate of aPKC-I was significantly higher in cholangiocarcinoma than in benign bile duct tissues （68.6% vs. 11.1%, P = 0.006）, while the positive rate of E-cadherin was significantly lower in cholangiocarcinoma than in benign bile duct tissues （37.1% vs. 88.9%, P = 0.016）. aPKC-I expression was negatively correlated to E-cadherin expression （r = -0.287, P 〈 0.05）. aPKC-I expression was positively and E-cadherin expression was negatively correlated to the differentiation and invasion of cholangiocarcinoma （P 〈 0.05）. Conclusion： The expressions of aPKC-I and E-cadherin may reflect the differentiation and invasive potential of cholangiocarcinoma. As a polar regulation-associated protein, aPKC-I may play an important role in the invasion and metastasis of cholangiocarcinoma.     

Vascular Endothelial Growth Factor Expression in Invasive Ductal Carcinoma of Breast

Objective： To detect the expression of VEGF and MVD count in invasive ductal carcinoma of breast to clarify the association of VEGF expression and MVD count with the clinicopathologic features. Methods： The expressions of VEGF, ER, PR, C-erbB-2 and MVD count in 88 cases of invasive ductal carcinoma of breast were examined by immunohistochemistry staining （SP-method）. Results： Sixty-two out of the eighty-eight specimens of breast carcinoma （70.45%） showed positive expression of VEGF. The positive rate of VEGF in cases with lymph node metastasis was higher than that without lymph node metastasis （P〈0.05）. The positive rate of VEGF in stage IIb-Ⅲ was higher than that in stage Ⅰ-Ⅱa （P〈0.05）. The positive rate of VEGF in C-erbB-2 positive group was higher than that in C-erbB-2 negative group （P〈0.05）. Higher expression of VEGF was observed in cases with higher tissue differentiation degree （P〈0.05）. Also, significant higher MVD count was observed in cases with higher tissue differentiation degree （P〈0.01）. The MVD count increased significantly with the increase of the expression of VEGF （P〈0.01）. Conclusion： The result of this study suggested that in invasive ductal carcinoma of breast, angiogenesis and metastasis were mediated mainly by VEGF. The expression of VEGF and MVD might be reference predictors for the biological behavior of breast carcinoma. The antiangiogenic therapy which used VEGF as a target would become a new method to treat patients who were C-erbB-2 positive in the future.     

Prognostic Value of P-gp and p27 in Patients with Esophageal Squamous Cell Carcinoma

Objective： To evaluate the prognostic value of P-gp and p27 expression in patients with esophageal squamous cell carcinoma （ESC）. Methods： The expressions of P-gp and p27 were detected by immunohistochemistry in 104 cases of ESC, and the clinicopathological characteristics were analyzed as well. Results： The positive rate of P-gp expression in 104 cases of ESCs was 32.7%. The positive rate of P-gp expression in the group that survived over 3 years （17.5%） was significantly lower than that in the group died within 3 years （53.3%） （x^2=14.227, P〈0.001）. The positive rate of p27 expression in 104 cases of ESCs was 67.3%. The positive rate of p27 expression in the group that survived over 3 years （75.8%） was significantly higher than that in the group died within 3 years （56.5%） （x^2=4.361, P〈0.05）. The patients with poorer differentiation whole wall invasion, lymph node metastasis and more advanced TNM stage had a shorter survival than did those with better differentiation, more superficial invasion, no lymph node involvement and earlier TNM stage; and it was statistically significant （P〈0.05）. However, tumor size, macropathologic type, age and gender had no prognostic impact on ESC patients （P〉0.05）. Conclusion： P-gp and p27 expression levels had a clinical prognostic significance in ESC. It could provide a reference basis for selecting the chemotherapy projection. The tumor differentiation degree, depth of invasion, lymph node involvement and TNM stages all were correlated to ESC patients＇ survival.     

金属硫蛋白在三种女性肿瘤内表达的临床意义

Objective： To study the expressions and significations of metallothionein （MT） in cervical squamous cell cancer （CSC）, bladder transitional cell cancer （BTC） and breast cancer （BC） of woman. Methods： Immunohistochemical method was used to examine the expresses rate of MT in three types of woman cancer tissue. Results： The expressions rates of MT were 54.35% （29146） in BTC, 67.05% （59188） in BC and 57.14% （40/70） in CSC. The positive rate of MT expression was higher in low differentiation group than well differentiation group in BTC and CSC （P 〈 0.05）. Positive of MT in lobular cancer was significance higher than medullary and duct cancers （P 〈 0.05）. Conclusion： The expression of MT is related to differentiation degree, and it is a guidance for clinical choice of chemotherapy project.     

Early age of onset, multiple primary malignancies and poor prognosis are indicative of an inherited predisposition to esophageal squamous cell carcinoma in familial rather than sporadic disease — An update based on 14- to 23-year follow-up

OBJECTIVE To demonstrate the effects of an inherited predisposition to familial esophageal squamous cell carcinoma （ESCC） through the comparison and analysis of the clinicopathologic differences between familial and sporadic ESCC cases. METHODS Differences in age of onset, prevalence rates of double primary ESCC, and survival rates between familial ESCC （n = 476） and sporadic ESCC cases （n = 1226） were analyzed. RESULTS Overall, familial ESCC cases showed a significantly younger age of onset （51.9±8.2 vs. 53.4 ±8.0, Pt.test = 0.00）, a significantly higher prevalence rate for double ESCC （2.73 % vs. 1.22%, adjusted with TNM：χMH2 = 4.029, P = 0.045）, and a lower survival rate than in sporadic cases （Pwald = 0.04）. The familial cases showed both a younger age of onset and poorer survival in most subgroups, and the differences were more marked in early-stage rather than in the .late-stage disease groups. CONCLUSION Theses findings confirm the existence of familial as opposed to sporadic ESCC. By the theory of the ＂two-hit＂ origin of cancer, these findings also suggest that the ＂first hit＂, a genetic predisposition, can affect the age of onset, number of primary carcinomas, and the prognosis for familial ESCC patients.     

热休克蛋白在食管鳞癌中的表达及其意义

目的探讨热休克蛋白（HSPs）HSP27、HSP60、HSPT0和HSP90d在食管鳞癌及其切缘正常食管黏膜中的表达及其意义。方法168例食管鳞癌和42例切缘正常食管黏膜制成组织芯片,应用免疫组化EnVision法及Westernbolt检测HSP27、HSP60、HSP70和HSP90α在组织中的表达情况,分析4种HSPs的表达与肿瘤位置、肿瘤长度、浸润深度、分化程度和淋巴结转移的关系。结果食管鳞癌和切缘正常食管黏膜中,HSP27的阳性率分别为62．0％和42．1％;HSP60的阳性率分别为92．7％和63．2％;HSP70的阳性率分别为57．9％和22．2％;HSP90α的阳性率分别为33．7％和18．5％。统计学分析结果表明,HSP60和HSP70在食管鳞癌的表达均高于切缘正常食管黏膜（P〈0．01）,HSP27和HSP90α在食管鳞癌与切缘正常黏膜的表达差异均无统计学意义（P〉0．05）。除HSP27的表达随着食管鳞癌分化程度的降低而降低（P〈0．05）外,其他3种HSPs的表达与食管鳞癌的临床病理特征无关（P〉0．05）。结论HSP27、HSP60、HSPT0和HSP90α在食管鳞癌和切缘正常食管黏膜中均有表达。HSP60和HSP70在食管鳞癌的表达高于切缘正常食管黏膜,可能与食管鳞癌的生物学行为有关;HSP27的表达随食管鳞癌分化程度的降低而降低,HSP27的高表达可能影响鳞癌的分化程度。     

食管鳞状细胞癌中热休克蛋白10和60的表达及其意义

目的探讨食管鳞状细胞癌中热休克蛋白(HSP)10、60的表达及其意义。方法应用免疫组化:EnVision二步法,观察168例食管鳞状细胞癌和42例切缘食管黏膜中HSP10、60的表达,并比较其阳性率。结果 HSP10在食管鳞状细胞癌和切缘食管黏膜表达率分别为53．8％和 37．5％,统计学分析结果表明差异无显著意义(P>0．05);HSP60在食管鳞状细胞癌和切缘食管黏膜表达率分别为92．7％和63．2％,统计学分析结果表明差异有显著意义(P<0．01)。在高、中、低分化食管鳞状细胞癌中HSP10、60的表达差异均无显著意义(P>0．05),两者的表达与食管鳞状细胞癌区域淋巴结转移无关(P>0．05)。结论 HSP60在食管鳞状细胞癌的表达高于切缘食管黏膜,提示其在食管鳞状细胞癌的发生发展中起作用。HSP10、60的表达与食管鳞状细胞癌分化程度、区域淋巴结转移无关。     

食管鳞癌组织中NK细胞和DC浸润与预后的关系

背景与目的自然杀伤(natural killer,NK)细胞和树突细胞(dendritic cell,DC)在抗肿瘤免疫中发挥重要作用.热休克蛋白(heat shock protein,HSP)也参与抗肿瘤细胞免疫.本研究旨在探讨食管鳞癌组织中NK细胞和DC浸润与预后的关系及其与HSP27、HSP70表达的关系.方法应用免疫组化SP法检测101例食管鳞癌组织中NK细胞和DC的浸润情况.根据临床病理资料和随访结果,对比分析NK细胞和DC在临床病理特征相同、但术后生存时间明显不同的两组病例(长期生存组术后生存5年以上,38例;短期生存组术后1年内死亡,63例)癌组织中的浸润情况.同时,分析NK细胞和DC浸润与HSP27、HSP70表达的关系.结果免疫组织化学染色结果显示101例食管鳞癌组织中每高倍视野(×400)NK细胞平均数为(31.4±19.7)个,其中长期生存组癌组织中每高倍视野NK细胞平均数为(41.9±21.7)个,明显高于短期生存组(25.0±15.4)个(P<0.001).DC在101例食管鳞癌组织中每高倍视野细胞平均数为(14.8±7.8)个,长期生存组和短期生存组癌组织中每高倍视野DC平均数分别为(15.2±8.6)个和(14.5±7.4)个,t检验显示两组无显著性差异(P>0.05).NK细胞和DC浸润与肿瘤长度、浸润深度、病理分级、淋巴结转移等临床病理特征均无相关性(P>0.05).二项分类logistic回归分析显示NK细胞被选入食管鳞癌预后模型(P=0.001),而DC未能进入该模型中(P=0.842).相关分析显示,NK细胞浸润和DC浸润之间呈正相关(r=0.266,P=0.007).NK细胞和DC的浸润与HSP27的表达呈正相关(r=0.237,P=0.017;r=0.235,P=0.018).NK细胞和DC的浸润与HSP70的表达无相关性(P>0.05).结论NK细胞浸润是食管鳞癌预后的独立影响因素,可作为判断食管鳞癌预后的重要生物指标之一.DC的浸润与食管鳞癌预后无关.NK细胞和DC的浸润与HSP27的表达可能有内在联系.     

p27Kip1、PLK1在食管鳞癌中表达的临床意义

目的:探讨p27Kip1(p27)、polo-like kinase1(PLK1)在食管鳞癌(ESCC)与其癌旁组织中的差异表达及其相关性,分析其基因表达差异与ESCC的临床病理特点及蛋白表达与预后的关系。方法:RT-PCR和免疫组化检测70对ESCC与其癌旁组织中p27、PLK1的mRNA和蛋白表达情况。结果:比较70对ESCC与其癌旁组织中p27及PLK1的相对表达,p27在46例(66%)癌组织中相对表达下降,24例(34%)癌组织中相对表达增高,比较有统计学差异(P＜0.01);而PLK1在63例(90%)癌组织中相对表达增高,7例(10%)癌组织中相对表达下降,比较有统计学差异(P＜0.01);且癌组织中PLK1及p27相对低表达与患者的病理进展、淋巴结的转移和临床分期相一致(P＜0.05);癌组织中PLK1伴p27胞浆表达阳性的患者与癌组织中PLK1伴p27胞核表达阳性的患者比较,术后生存期明显缩短(P＜0.01)。结论:p27与PLK1在ESCC组织中的差异性表达与患者的临床病理特点密切相关,癌组织中PLK1伴胞浆p27表达阳性的患者预后更差。     

HSP27 and HSP70 Expression in Esophageal Squamous Cell Carcinoma

Twenty-eight specimens of Esophael squamous cell carcinoma (ESCC) were obtained by surgery procedures.The tissues were fixed in formalin and embedded in paraffin. In each case, all available hematoxylin and eosin stained sections were examined and a representative block was selected. The ages of these patients ranged from 40 to 93 years, with a mean age of 60 years. Results. The histological grade of tumors was 4 well-differentiated, 19 moderately differentiated and 5 poorly differentiated. Expression of Hsp27 and Hsp70 in ESCC was demonstrated in 23 (82,14%) and 26 (92,86%) cases, respectively. Adjacent normal mucosa was positive in 11 (39,29%) samples and 9 (32,15%) samples for Hsp27 and Hsp70, respectively. No relationship between the expression of Hsp27 and Hsp70 with the clinicopathological parameters, including gender, age, surgical margin, lymph node status and tumor differentiation. The median follow-up period was 60 months. Survival analysis of patients with ESCC showed no relationship with the expression of Hsp27 and Hsp70. Conclusion. Taken together, our results demonstrate that Hsp27 and Hsp70 are expressed in ESCC tissues, but they are not good prognostic factor for patients with ESCC.     

食管鳞癌组织中RECK mRNA和蛋白的表达及意义

目的:探讨食管鳞癌组织中RECK mRNA和蛋白的表达情况及其与临床病理因素的关系.方法:应用RT-PCR和免疫组化SP法检测62例食管鳞癌组织、31例癌旁不典型增生组织及62例正常食管粘膜组织中mRNA和蛋白的表达.结果:在食管鳞癌癌变过程中RECK在癌组织、癌旁不典型增生组织及正常粘膜组织中mRNA的含量依次增高,分别为1.052±0.078、1.274±0.235、1.306±0.121,组间比较有明显差异(F=49.936,P＜0.05):不同分化程度、不同浸润深度及有无淋巴结转移的食管鳞癌组织之间RECK mRNA相对含量差异均有统计学意义(F=5.081,F=26.084,U=24.011,P均＜0.05).食管鳞癌组织及癌旁不典型增生组织中RECK蛋白表达均低于正常粘膜组织,表达量分别为59.7%(37/62)、71.0%(22/31)、85.5%(53/62),组间比较有明显差异(x2=10.331,P＜0.01);食管鳞癌组织中RECK蛋白表达与癌组织的分化程度、不同浸润深度及有无淋巴结转移密切相关(P＜0.05).结论:食管鳞癌组织中RECK mRNA和蛋白表达均降低,其低表达可能与食管鳞癌发生有关.检测RECK mRNA及蛋白的表达可望成为食管鳞癌早期诊断和判断预后的分子指标之一.     

细胞色素P450 2E1和畸胎瘤细胞源性生长因子在食管鳞状细胞癌中的表达及临床意义

目的 探讨食管鳞状细胞癌（ESCC）中细胞色素P450 2E1（CYP2E1）、畸胎瘤细胞源性生长因子（PCDGF）的表达与肿瘤临床病理参数之间的关系.方法 选择郑州市第一人民医院外科2007年7月至2011年5月间手术切除ESCC标本42例及20例癌旁正常组织.采用免疫组织化学方法检测食管鳞状上皮中C YP2E1与PCDGF的表达.结果 ESCC中CYP2E1、PCDGF的阳性表达率分别为83.3％（35/42） 、88.1％（37/42）,均较正常食管上皮明显增加（P＜ 0.05）;PCDGF与肿瘤的组织分化程度（r=0.444,P＜ 0.05）、浸润深度（r=0.332,P＜0.05）、淋巴结转移（r=0.476,P＜0.05）和TNM分期（r=0.450,P＜ 0.05）有关;CYP2E1的表达与肿瘤的组织分化程度呈负相关（r=-0.518,P＜0.05）,而与浸润深度呈正相关（r=0.388,P＜ 0.05）.CYP2E1和PCDGF在肿瘤中的表达有相关性（r=0.483,P＜ 0.05）.结论 ESCC中CYP2E1和PCDGF相互协同参与了肿瘤生长、浸润和转移,可能成为判断食管癌生物学行为及患者预后的重要指标.     

流式细胞术对食管癌组织中四种细胞周期调控蛋白的定量检测及意义

目的 探讨细胞周期调控蛋白cyclin E、cyclin D1、CDK4和D27在食管癌的表达及其与食管癌的分化和淋巴结转移的关系。方法 采用流式细胞术对65例食管鳞状细胞癌组织中cyclin E、cyclin D1、CDK4和p27的表达强弱进仃定量检测,结果 用荧光指数F1表示。结果 cyclin E、cyclin D1和CDK4在低分化型鳞癌的表达量显著高于分化型鳞癌(P值分别为0．0275,0．0001和0．0174);而p27在低分化型鳞癌的表达量显著低于分化型鳞癌(P=0．0042)。cyclin D1与cyolin E,cyclin D1与CDK4之间呈显著正相关;而cyclin D1与p27呈显著负相关。4种基因蛋白的表达与淋巴结转移均无相关性。结论 cyclhi E、cyclin D1、CDK4和p27的表达与食管癌的分化密切相关;正负性细胞周期调控蚩白表达的失衡是导致癌变的重要原因之一。     

PLK4基因在人食管鳞癌组织中的表达及对癌细胞增殖、侵袭迁移影响的研究

背景与目的：作为调节细胞周期的蛋白激酶,polo样激酶4（polo-like kinase 4,PLK4）参与有丝分裂启动、中心体成熟、胞质分裂及DNA损伤检测等,在多种肿瘤中呈高表达,但PLK4是否参与食管鳞状细胞癌（esophageal squamous cell carcinoma,ESCC）的增殖和侵袭迁移尚不清楚。研究PLK4在ESCC细胞系和临床组织标本中的表达以及对癌细胞增殖、侵袭迁移的影响。方法：采用TRIzol提取细胞总RNA,反转录试剂盒合成cDNA,采用实时荧光定量聚合酶链反应（real-time fluorescent quantitative polymerase chain reaction,RTFQ-PCR）检测PLK4基因在正常食管上皮细胞Het-1A和ESCC细胞系TE-1、TE-8和TE-13中的mRNA表达水平。离心收集细胞后用RIPA裂解液提取细胞总蛋白,采用蛋白质印迹法（Western blot）检测正常食管上皮细胞Het-1A和ESCC细胞系TE-1、TE-8和TE-13中PLK4蛋白水平。选取2017年1月—2019年12月河南医学高等专科学校附属医院93例经病理组织学检查确诊为ESCC的患者癌组织及其配对的癌旁组织（距原发灶边缘5 cm以上）,临床组织用液氮速冻后加入RIPA裂解液研磨提取组织总蛋白,采用Western blot检测PLK4蛋白水平。绘制受试者工作特征曲线,分析PLK4表达与临床病理学参数之间的关系。利用siRNA干扰技术抑制TE-13细胞中PLK4的表达,设计并合成PLK4的siRNA干扰片段,采用Lipofectamine ^TM 2000转染细胞抑制PLK4在TE-13细胞中的表达,RTFQ-PCR实验和Western blot检测siRNA干扰片段对PLK表达的影响。PLK4在TE-13细胞中表达下调后,用细胞计数试剂盒-8（cell counting kit-8,CCK-8）实验和克隆形成实验检测细胞的增殖能力,transwell小室实验检测细胞的侵袭能力,划痕愈合实验检测细胞的迁移能力。Western blot实验检测PLK4下调后对mTOR/p70S6K信号转导通路中关键蛋白mTOR、p70S6K、p-mTOR ^Ser2448 和p-p70S6K ^{Thr421/Ser424} 表达的影响。结果：RTFQ-PCR和Western blot检测结果显示,PLK4基因在ESCC细胞系中的mRNA和蛋白表达水平显著高于正常食管上皮细胞（P＜0.05）。与癌旁正常组织相比,ESCC组织标本中PLK4的蛋白表达水平异常增高（P＜0.05）,绘制的受试者工作特征曲线的曲线下面积为0.841,95%CI为0.786~0.895,灵敏度为74.2%（69/93）,特异度为89.2%（83/93）（P＜0.000 1）。PLK4蛋白在ESCC组织中的表达水平与性别、年龄和肿瘤大小均无关（均P＞0.05）,但与分化程度、临床分期和淋巴结是否转移有关（均P＜0.05）。ESCC分化程度越低,PLK4高表达率越高,低分化程度的ESCC组织中PLK4高表达率为86.7%,Ⅲ~Ⅳ期患者ESCC组织中PLK4蛋白高表达率为92.3%。PLK4高表达与临床分期呈正相关（P＜0.05）。CCK-8和克隆形成实验结果显示,下调PLK4的表达可以显著抑制TE-13细胞的增殖（P＜0.05）,transwell小室实验和划痕实验结果显示,下调PLK4的表达可以显著抑制TE-13细胞的侵袭迁移能力（P＜0.05）。抑制PLK4的表达使TE-13细胞中mTOR和p70S6K蛋白的表达下降（P＜0.05）,且p-mTOR ^Ser2448 和p-p70S6K ^{Thr421/Ser424} 的表达下降（P＜0.05）。结论：PLK4在ESCC细胞和组织中均呈高表达,抑制PLK4的表达可以抑制ESCC细胞的增殖和侵袭迁移能力,PLK4可能通过mTOR/p70S6K信号转导通路促进ESCC细胞恶性进程。