Comparative Proteome Analysis of Human Lung Squamous Carcinoma Tissue

目的利用蛋白质组学方法建立人肺鳞癌组织及其癌旁正常支气管上皮组织的差异蛋白质表达谱。方法对20例人肺鳞癌组织和配对的癌旁正常支气管上皮组织进行比较蛋白质组学研究,即利用双向凝胶电泳(2-DE)分离二者总蛋白质后,经图像分析识别差异表达的蛋白,应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)鉴定差异蛋白质。结果(1)比较分析20例肺鳞癌及正常配对组织的2-DE图谱,找到差异蛋白质点76个;(2)对68个差异蛋白质点进行了肽质指纹图分析,鉴定出一些与瘤基因、细胞周期调控、信号转导等有关的肺鳞癌相关蛋白;(3)肺鳞癌相关蛋白mdm2、c-jun和EGFR存人肺鳞癌组织中高表达,而在正常对照中均表达下调,与蛋白质组的分析鉴定结果是一致的。结论成功鉴定了68个肺鳞癌相关蛋白,为进一步筛选用于肺鳞癌诊断、治疗和预后评估的肺鳞癌分子标志物奠定了坚实的基础。     

人肺鳞癌组织的比较蛋白质组学研究

目的利用蛋白质组学方法建立人肺鳞癌组织及其癌旁正常支气管上皮组织的差异蛋白质表达谱。方法对20例人肺鳞癌组织和配对的癌旁正常支气管上皮组织进行比较蛋白质组学研究,即利用双向凝胶电泳（2-DE）分离二者总蛋白质后,经图像分析识别差异表达的蛋白,应用基质辅助激光解吸电离飞行时间质谱（MALDI—TOF—MS）鉴定差异蛋白质。结果（1）比较分析20例肺鳞癌及正常配对组织的2-DE图谱,找到差异蛋白质点76个;（2）对68个差异蛋白质点进行了肽质量指纹图分析,鉴定出一些与瘤基因、细胞周期调控、信号转导等有关的肺鳞癌相关蛋白;（3）肺鳞癌相关蛋白mdm2、c-Jun和表皮生长因子受体（EGFR）在人肺鳞癌组织中高表达,而在正常对照中均表达下调,与蛋白质组的分析鉴定结果是一致的。结论成功鉴定了68个肺鳞癌相关蛋白,为进一步筛选用于肺鳞癌诊断、治疗和预后评估的肺鳞癌分子标志物奠定了坚实的基础。     

Analysis of serum proteome profiles in nasopharyngeal carcinoma with different radiosensitivity

Purpose

The research is endeavored to establish a serum protein fingerprint model for predicting radiosensitivity of nasopharyngeal carcinoma through the analysis of the serum expressed proteins of the pre- and post-treatment nasopharyngeal carcinoma patients treated with radiotherapy.

Methods

Surface-enhanced laser desorption ionization-time of flight-mass spectrometry and CM-10 protein chip were used to detect the serum proteomic patterns of 50 nasopharyngeal carcinoma patients with different radiosensitivity. Thirty-eight of the 50 patients after the treatment were also studied. Biomarker Wizard 3.01 and Biomarker Pattern 5.01 were used in combination to analyze the data and to develop predicting models.

Results

At the molecular weight range of 2,000–20,000, the software identified an average of 83 mass peaks between radiation sensitive group and radiation resistant group. 11 protein peaks were significantly different. Of the 83 mass peaks, 4 mass peaks (mass/charge ratio [m/z] 2,575, 3,942, 6,117 and 6,778) were chosen automatically to construct a classification tree. The diagnostic accuracy was 78.0 %. M/z 6,117 and 6,778 of the radiation sensitive group after the treatment trended to those of the radiation resistant group.

Conclusions

The technology of surface-enhanced laser desorption ionization-time of flight-mass spectrometry can be used to screen and identify differentially expressed proteins associated with different radiosensitivity in nasopharyngeal cancer. It should be a very useful tool for predicting radiosensitivity of nasopharyngeal carcinoma.     

Improved survival of patients with nasopharyngeal squamous cell carcinoma

One hundred and one patients with nasopharyngeal squamous cell carcinoma (NPC) were treated with irradiation. The UICC TNM staging system (1978) was used: 6 patients were T1N0, 10 T2N0, 5 T3N0, 13 T4N0, 11 T1N+, 18 T2N+, 19 T3N+ and 19 T4N+. Since 1978, 34 patients were examined with computed tomography (CT) at first presentation. Fourteen (41%) of the 34 tumors were upstaged based on the CT findings alone. The technique of radiation therapy was markedly changed around 1978. The fields to the primary site and neck were enlarged. Two-year relapse-free survival was significantly better for the post-CT era than pre-CT era. This was mainly because of improved local-recurrence-free survival, and cervical-relapse-free survival. Improved local-recurrence-free survival, however, was appreciated in only T3 + T4 patients; there was no difference in T1 + T2 patients. It is suggested that merely enlarging radiation fields or increasing radiation doses could not be curative for some subpopulations. In order to increase local control rates further, we have started to use intracavitary irradiation with an after-loading technique as a boost. Preliminary results are encouraging.     

Deep proteome profiling promotes whole proteome characterization and drug discovery for esophageal squamous cell carcinoma

Proteins are key players in various cellular processes. As the ultimate executors of cellular processes, they play essen-tial roles in linking genotypes to phen...     

宫颈癌患者血清蛋白指纹图谱的检测及其意义

背景与目的:目前针对宫颈癌没有特异性的肿瘤标志物。表面增强激光解吸离子化飞行时间质谱(surface-enhanced laser desorption/ionization time-of-flight mass spectrometry,SELDI-TOF-MS)是最新应用的一项检测肿瘤标志物的技术。本研究应用SELDI-TOF-MS检测宫颈癌患者血清蛋白指纹图谱,筛选候选肿瘤标志物并建立诊断模型,初步探讨其在宫颈癌早期诊断中的价值。方法:取91例早期宫颈鳞癌患者和15例宫颈上皮内瘤变Ⅲ级(cervical intraepithelialneoplasia,CINⅢ)患者的血清标本进行实验,同时用55名健康人血清作为对照。用弱阳离子交换芯片(weak cation exchange,WCX2)检测各血清标本获得血清蛋白指纹图谱。用Biomarker Patterns软件分析宫颈癌差异蛋白并建立诊断模型。通过盲法分析进一步验证诊断模型的可靠性,并对结果进行统计学分析。结果:在分子量1.5～20ku范围内,共检测到122个蛋白峰,其中19个差异峰有统计学意义(P<0.001)。建立了由分子量为3977和5807的两个差异蛋白组成的宫颈癌诊断模型,其敏感性为97.29%(36/37),特异性为83.78%(31/37)。扩大样本盲法验证结果,其敏感性为94.44%(51/54),特异性为94.44%(17/18)。结论:由3977和5807两个差异蛋白组成的宫颈癌诊断模型有助于区分宫颈癌和健康人群。     

A human squamous cell carcinoma cell line.

An epithelial cell line COLO 16 has been established from a human squamous carcinoma, characterized and maintained for over two years. The cells produce a parathyroid-like hormone and carcinoembryonic antigen. The line is definitely not a "HeLa contaminant." The cell line is available to other investigators.     

高转移能力与低转移能力人鼻咽癌细胞株蛋白质组学比较分析

目的:采用蛋白质组学方法对比分析高转移能力与低转移能力鼻咽癌细胞株,寻找差异表达蛋白质。方法:采用双向凝胶电泳技术分析高转移能力鼻咽癌细胞系S-18和低转移能力细胞系S-22,寻找S-18与S-22蛋白质表达差异。采用傅立叶变换离子回旋共振质谱技术,结合生物信息学数据库检索,鉴定出差异表达的蛋白质。结果:鉴定出12种差异表达蛋白质,包括转酮酶、NSFL1(p97)辅因子(p47)、磷酸肝油酸激酶1、醛缩酶A、核纤层蛋白A/C、膜联蛋白A2、SKP1G2等位基因抑制因子、不均一核核糖核蛋白C(C1/C2)、动力蛋白轻链Tctex型1、硫氧还蛋白、桩蛋白、丝氨酸/苏氨酸激酶受体关联蛋白。结论:这12种差异表达在鼻咽癌转移能力中可能起到关键性作用,为进一步研究鼻咽癌转移机制、分子标志及治疗靶点提供研究方向。     

人鼻咽与鼻咽癌cDNA阵谱中DNA修复相关基因表达差异的初步研究

目的 用cDNA阵卤咽癌组织及正常组织的基因表达谱,研究鼻咽癌组织内修复相关基因的表达差异。方法 Atlas human cancer cDNA expression array 7742-1杂交后,用Atlas Image 1.01a分析滤膜杂交结果,RT－PCR反应验证滤膜杂交结果。结果 在588个肿瘤相关基因中,共有134个基因表达上调,88个基因表达下调。在有DNA损伤应答、修复、重组相关基因的C区中,表达上调的基因有21个,与修复相关的有6个;表达下调的有44个,与修复相关的则有12个。结论 DNA修复相关基因的改变可能在鼻咽癌病理生理过程中起一定作用。     

基因甲基化抑制鼻咽癌组织SCCA1的表达

背景与目的:鳞状细胞癌抗原1(squamous cell carcinoma antigen 1,SCCA1)与鳞状细胞癌的生长、分化及转移密切相关,鼻咽癌(nasopharyngeal carcinoma,NPC)组织SCCA1表达水平低于鼻咽黏膜上皮组织,其表达降低的机制不清楚.本研究初步探讨NPC组织SCCA1表达水平降低是否由基因甲基化所致.方法:收集75例NPC活检组织(NPC组)和25例慢性鼻咽炎鼻咽黏膜活检组织(对照组),采用甲基化特异性聚合酶链式反应检测SCCA1基因甲基化状态,PCR检测SCCA1 mRNA表达水平,免疫组织化学染色和Western blot检测SCCA1蛋白表达情况.结果:NPC组SCCA1基因完全甲基化、不完全甲基化和未甲基化的例数分别为38例、27例和10例,对照组不完全甲基化和未甲基化的例数分别为5例和20例,NPC组甲基化频率显著高于对照组(65/75vs 5/25,x2=39.7,P＜0.01).SCCA1基因不完全甲基化导致mRNA和蛋白表达降低,SCCA1基因完全甲基化导致mRNA和蛋白表达缺失.在NPC组,SCCA1不完全甲基化标本mRNA表达水平和蛋白表达水平显著低于SCCA1未甲基化病例(0.22±0.05 vs 0.37±0.07和0.14±0.05 vs 0.31±0.04,P＜0.01);等级相关分析显示SCCA1基因甲基化程度与NPC组织免疫组织化学染色积分呈负相关(P＜0.01),与NPC淋巴结转移分期呈正相关(P＜0.01).结论:NPC组织SCCA1蛋白质表达降低主要是由基因甲基化所致,SCCA1表达降低可能对NPC淋巴结转移具有促进作用.     

Molecular pathology parameters in human nasopharyngeal carcinoma

BACKGROUND: To derive a better understanding of the biologic behavior of nasopharyngeal carcinoma (NPC), the authors evaluated a number of molecular variables to address the hypothesis that p53 dysfunction in NPC is associated with Epstein-Barr virus (EBV), increased tumor angiogenesis, lower likelihood of apoptosis, and poorer clinical outcome. MATERIALS: The biopsy samples from 87 NPC patients were obtained and sections were made to detect EBV, using in-situ hybridization; the authors used immunohistochemistry to assess p53, p21(WAF1/CIP1) expression, and microvessel density count (MVD). In situ end labelling was used to evaluate apoptosis and necrosis. Analyses were conducted on the association between each of these variables as well as clinical outcome, including survival and local control. RESULTS: There was a highly significant association between EBV-encoded RNA (EBER) positivity with p53 over-expression in that only 1 out of 32 p53 over-expressing tumors was EBER negative, as opposed to 19 out of 48 p53 negative tumors being EBER negative (P = 0.001). In addition, EBER positivity was highly associated with World Health Organization (WHO) type 3 NPC, Asian/Chinese ethnicity, a lower apoptotic index, and p21 over-expression. p53 over-expression was associated with a higher MVD count. Controlling for age and nodal status, EBER positivity was associated with both improved overall survival (P = 0.02), and disease-free survival (P = 0.04). In contrast, the presence of tumor necrosis was associated with an inferior local control (P = 0.03). CONCLUSION: p53 protein was over-expressed in approximately one third of NPC samples in the current study, and this correlated significantly with the presence of EBER. Epstein-Barr virus status was also associated with WHO type 3 NPC, Asian/Chinese ethnicity, and induction of p21. The presence of EBV appeared to predict for improved survival, the mechanism of which remains to be elucidated in this biologically complex disease.     

乙醛脱氢酶1在鼻咽鳞状细胞癌中的表达及其临床意义

目的 研究乙醛脱氢酶1（ALDH1）在鼻咽鳞状细胞癌中的表达及其临床意义.方法 采用免疫组织化学方法检测确诊为鼻咽鳞状细胞癌未行放化疗的120例患者石蜡标本中ALDH1的表达情况.结果 ALDH1在40.0％（48/120）鼻咽鳞状细胞癌患者中呈中、高表达,在癌巢和间质的边缘,尤其在梭形细胞中呈高表达.ALDH1表达与鼻咽原发灶范围（P=0.011）、颈淋巴结转移（P=0.005）、临床分期（P=0.001）相关,而与年龄、性别无关（均P＞0.05）.Kaplan-Merier生存分析和Cox回归分析均表明ALDH1表达与鼻咽鳞状细胞癌患者的临床预后相关,并且ALDH1是鼻咽鳞状细胞癌患者临床预后的独立危险因素（HR=2.056,P＜0.05）.结论 鼻咽鳞状细胞癌ALDH1的表达与患者的临床预后密切相关,是影响其临床预后的危险因素之一.     

鼻咽癌放疗后第二原发舌鳞癌35例

 目的　探讨鼻咽癌放疗后第二原发舌鳞癌的临床特点、有效的治疗手段、生存情况和影响预后的因素。方法　回顾性分析35例鼻咽癌放疗后第二原发舌鳞癌患者的临床和随访资料,应用Kaplan-Meier法及Log-Rank检验、COX回归模型等对本组病例资料进行统计分析。结果　35例患者总的3、5年生存率分别为55 %、47 %,淋巴结转移率为5.71 %;单因素分析提示性别（χ2＝8.89,P＝0.00）、T分级（χ2＝5.58,P＝0.02）、临床分期（χ2＝8.51,P＝0.04）、治疗方法（χ2＝29.37,P＝0.00）是影响预后的重要因素;多因素分析显示治疗方法（P＝0.00）、T分级（P＝0.03）是与预后有关的重要独立因素;手术治疗组较非手术治疗组预后好,差异有统计学意义（P <0.05）;男性患者发生第二原发舌鳞癌的风险较女性患者高;第二原发舌鳞癌随鼻咽癌放疗后时间的延长发病率升高。结论　鼻咽癌放疗后第二原发舌鳞癌淋巴转移率较低;治疗方法和T分期是影响预后的独立因素;鼻咽癌放疗后需长期随访,早期诊断第二原发舌鳞癌,并给予手术或包含手术的综合治疗,以取得好的疗效。     

Treatment of nasopharyngeal carcinoma with human leukocyte interferon

Nasopharyngeal carcinoma (NPC) is a human neoplasm closely associated with Epstein-Barr virus (EBV). Human leukocyte interferon (IFN) has known antiviral and antineoplastic properties. After initial IFN treatment in one NPC patient demonstrated acceptably low toxicity, 12 additional patients were treated on a protocol with IFN, 10 X 10(6) units intramuscularly (IM) daily for 30 days. IFN did not affect serum anti-EBV antibody titers (IgA and IgG antiviral capsid and early antigens). Of six patients tested, none was found to excrete EBV in saliva before, during, or after IFN. Four patients had measurable tumor regression (two partial responses and two minor responses), three had stable disease, and five patients plus the initial preprotocol patient had progressive disease. Toxicity included fever, fatigue, and myalgias in all patients, thrombocytopenia in two patients, and neutropenia in three patients. Three patients were withdrawn from the study, one each for severe fatigue, neutropenia, and hypotension. This study demonstrates that IFN has sufficient activity in advanced NPC to justify further investigation.     

Characterization of radiation sensitivity of human squamous carcinoma A431 cells

Studies have been performed to investigate the radiosensitivity of human squamous carcinoma cells. A431 cells were grown in vitro as exponential and fed-plateau monolayer cultures or as multicellular spheroids. Radiobiological studies of various cultures showed that fed-plateau phase cells were more sensitive (D0 = 1.3 Gy) than exponentially growing cells (D0 = 1.5 Gy). After a single dose of 12 Gy or two doses of 6 Gy irradiation, A431 cultures exhibited a large capacity for potentially lethal damage (PLD) repair (PLD repair factor = 17), but a relatively small sublethal damage (SLD) repair. In order to measure the radiation sensitivity of proliferating (P) and quiescent (Q) cells, enriched populations of P- and Q-cells were isolated from A431 spheroids. Flow cytometric analysis with acridine orange (AO) staining demonstrated that there was a shift of the RNA histograms in fed-plateau and spheroid cultures towards lower values, suggesting the presence of a subpopulation of Q-cells. Centrifugal elutriation was used to isolate the Q-cells from dissociated spheroid cells. Coulter cell volume distributions and flow cytometric analysis showed that Q-cells had a small cell volume (approximately 1380 microns3), low RNA content and a G1-like DNA content. Continuous labelling experiments with tritiated thymidine confirmed the non-proliferating nature of the Q-cells. Irradiation of the Q-cells after isolation from spheroids with between 0 to 10 Gy showed that they were more radiosensitive (decreased D0) than the P-cells isolated from these spheroids. The latter were, however, similar in radiosensitivity to exponential G1 cells.     

SCC在鼻咽癌患者血清中表达的临床意义

目的 探讨鼻咽癌患者血清中鳞状细胞癌抗原(SCC)的表达及临床意义.方法 用酶联免疫吸附法分析186例鼻咽癌患者检测血清SCC水平,并结合临床资料分析该指标的临床意义.结果 鼻咽癌患者SCC阳性率治疗前为29.03%,临床分期之间阳性率差别有统计学意义(P＜0.05),治疗后阳性率均表现下降,总的阳性率为7.0%.结论 SCC作为早诊手段,其敏感性较低,但在初诊鼻咽癌中的阳性率随病期增高,对病情预后监测有临床意义.     

食管鳞状细胞癌淋巴结转移与血清蛋白指纹图谱的相关性研究

目的 分析食管鳞状细胞癌(ESCC)患者血清蛋白表达谱的改变;比较有无淋巴结转移(LM)的患者血清蛋白指纹图谱,筛选与LM相关的蛋白峰.方法 术前收集64例ESCC患者和60名性别、年龄相匹配的健康成年人的血清.采用弱阳离子交换型(WCX-2)蛋白芯片为检查介质,经SELDI-TOF-MS测定蛋白指纹图谱.行有癌和无癌的对照研究;ESCC患者组内分别按性别、年龄、肿瘤位置、肿瘤大小、浸润程度和有无LM分别进行对照,得到蛋白指纹图谱并用Biomarker Wizard软件分析比较.结果 在质量/电荷(m/z)0～50000范围内,ESCC与健康成年人血清检测出120个蛋白峰,其中31个蛋白峰差异有统计学意义(P<0.05);不同浸润程度分组对比,T1与T3、T4比较有1个m/z是4174的差异蛋白峰(P<0.05);有无LM组间,3个蛋白峰差异有统计学意义(P<0.05),m/z分别为3970、4174、和4277.其中m/z为4174的蛋白峰是浸润组和LM组所共有的;年龄、性别、肿瘤位置、肿瘤大小分组未发现差异蛋白峰.结论 ESCC患者组和健康成年人组比较,血清蛋白有显著差异;ESCC浸润程度的不同和有无LM血清蛋白指纹存在一定差异,但远较ESCC与健康成年人之间差异小;浸润程度严重和LM在血清蛋白指纹中存在着一定的共性.