人大肠腺瘤和腺癌之间相关差异蛋白的蛋白质组学研究

目的　应用蛋白质组学的方法,建立人大肠腺癌组织、大肠腺瘤组织、正常大肠黏膜组织的二维电泳图谱,筛选并鉴定三者之间的差异蛋白,发现人大肠癌的潜在生物学标志物。方法　收集同时患有大肠腺瘤的大肠腺癌患者１０例,术前及术后病理证实均为Ｄｕｋｅｓ’Ｂ期。取同一患者术后大肠癌组织、距大肠癌组织１５ｃｍ以上正常大肠黏膜组织以及肠镜下切除大肠腺瘤组织,提取不同组织总蛋白进行二维电泳,用ＩｍａｇｅＭａｓｔｅｒ２ＤＰｌａｔｉｎｕｍ５.０凝胶图像分析软件对双向电泳图依次进行凝胶点的检测、背景消减和编号。识别不同组织之间的差异蛋白点,对差异蛋白点用胶内酶解后行质谱分析和网上数据库鉴定。结果　大肠腺癌组织、大肠腺瘤组织和正常大肠黏膜组织之间存在９０个差异蛋白点。对这些差异点进行质谱分析,经数据库鉴定出７１个蛋白质。经分析,大肠腺瘤组织与正常大肠黏膜组织相比,１７个蛋白表达上调,９个蛋白表达下调;大肠腺癌组织与正常大肠黏膜组织比较,４７个蛋白表达上调,１２个蛋白表达下调;大肠腺癌组织与大肠腺瘤组织相比,３５个蛋白表达上调,８个蛋白表达下调。结论　大肠腺癌组织、大肠腺瘤组织和正常大肠黏膜组织之间存在差异蛋白表达,可能与大肠腺癌的发生发展有关;应用蛋白质组学方法筛选人大肠腺癌发生发展相关蛋白是可行的。     

环氧合酶-2、前列腺素E2在大肠腺瘤和腺癌组织中的表达

目的:研究COX-2、PGE2在大肠腺瘤和腺癌组织中的表达和意义。方法:选取组织标本118例,其中正常大肠黏膜30例,大肠腺瘤43例,大肠腺癌组织45例。Real Time PCR法检测组织中COX-2mRNA的表达,免疫组织化Elivision法检测组织中COX-2的蛋白表达,放射免疫法测定组织中PGE2含量。结果:COX-2在大肠腺癌组织中的mRNA的表达、阳性表达率及光密度值分别高于大肠腺瘤及正常黏膜组(P<0.05);且大肠腺瘤组高于正常黏膜组(P<0.05)。大肠腺癌组织PGE2含量高于大肠腺瘤及正常大肠黏膜(P<0.01);且大肠腺瘤高于正常大肠黏膜(P<0.01)。结论:大肠黏膜癌变过程中COX-2表达增高,PGE2合成增多。     

S100A4蛋白在结肠腺瘤和结肠癌中的表达

[目的]探讨S100A4蛋白在结肠腺瘤和结肠癌中的表达及其临床意义。[方法]用免疫组化法检测S100A4在15例正常肠黏膜、80例结肠腺瘤和60例结肠癌中的表达。[结果]S100A4蛋白在正常肠黏膜中无表达,在结肠腺瘤中阳性率为16.25%,在结肠癌中阳性率为56.67%。[结论]S100A4与结肠腺瘤和结肠癌的发生发展密切相关,是预测结肠肿瘤进展的有用指标。     

应用蛋白质组学方法筛选结肠癌相关蛋白质

目的通过比较结肠癌组织与癌旁正常肠组织的差异表达蛋白谱,以期发现结肠癌癌变相关蛋白。方法将结肠癌组织与癌旁正常结肠组织蛋白进行双向凝胶电泳,选择在癌组织中高表达的50个点进行MALDI-TOF质谱分析和生物信息学分析。Western Blot和免疫组化方法验证蛋白的差异表达。结果建立了结肠癌及正常肠组织的双向凝胶电泳图谱,其中癌旁正常结肠组织和结肠癌组织电泳图谱中平均蛋白质点数分别为860±45和980±28;癌旁正常结肠组织和结肠癌组在IEF方向上的平均偏差分别为(0.487±0.11)mm和(0.654±0.11)mm,在SDS-PAGE方向上的平均偏差分别为(0.944±0.12)mm和(1.20±0.22)mm,结肠癌与癌旁正常结肠组织的差异表达蛋白质点数为72.00±12.34。选择在癌组织中高表达的50个点进行质谱分析和生物信息学查询,鉴定了其中有意义的25个点,包括谷胱苷肽S转移酶、肝脂肪酸结合蛋白、热休克蛋白27等。免疫组化方法检测结肠癌组织芯片中GST和HSP27的表达,结果显示,GST在结肠癌中表达的阳性率为73.7%,在正常肠黏膜中表达的阳性率为28.6%,差异有显著性(P<0.01);HSP27在结肠癌中表达的阳性率为73.7%,在正常肠黏膜中表达的阳性率为21.4%,差异有显著性(P<0.01)。结论蛋白质组学方法较好地显示结肠癌组织和癌旁正常组织的差异表达蛋白;其中在结肠癌组织中高表达蛋白HSP27和GST可能作为结肠癌诊断标志物。     

MAGE-A4在人大肠肿瘤中的表达及其与淋巴结转移的关系

目的研究MAGE-A4在人大肠肿瘤组织中的表达，并探讨其与大肠腺癌侵袭和转移的关系。方法采用免疫组织化学技术SP法，观察38例正常大肠黏膜、31例大肠腺瘤、83例大肠腺癌组织中MAGE-A4蛋白的表达特点。结果MAGE-A4蛋白的表达定位于胞质及胞核。在正常大肠黏膜组织、大肠腺瘤、大肠腺癌组织中的阳性表达率分别为5．26％、38．71％及38．55％。正常大肠黏膜组织中的阳性表达率与在大肠腺瘤、大肠腺癌组织中的阳性表达率相比较，有非常显著性差异（P〈0．01）；而在大肠腺瘤中的表达强度与在大肠腺癌中的相比，无显著性差异（P〉0．05）；在不同组织学类型的大肠腺瘤中的表达也无显著性差异（P〉0．05）；不同性别、不同年龄、不同侵袭深度、有肝转移与无肝转移的大肠腺癌患者，其大肠肿瘤组织中MAGE-A4蛋白的表达均无显著性差异（P〉0．05）；但有淋巴结转移与无淋巴结转移的大肠癌患者，其MAGE-A4蛋白的表达有显著性差异，且呈正相关关系（P〈0．01，γs=0．312）。结论MAGE-A4蛋白在大肠肿瘤的发生中有一定作用，且与大肠腺癌患者有无淋巴结转移有正相关关系。     

构建血清蛋白质质谱模型判别家族性腺瘤性息肉病与散发性肠腺瘤

目的 筛选家族性腺瘤性息肉病(FAP)的特异表达蛋白,构建判别FAP与散发性肠腺瘤的血清蛋白指纹图谱诊断模型.方法 采集19例FAP和16例散发性肠腺瘤患者的血清,以表面增强激光解吸电离飞行时间质谱仪(SELDI-MS-TOF)和阴离子CM01蛋白质芯片检测并筛选两组对象间的血清差异表达蛋白质峰,以支持向量机方法构建判别模型.结果 FAP与散发性肠腺瘤相比,P<0.01的蛋白质峰有6个,其中质荷比为5640、3160、4180和4290的蛋白质峰在FAP中高表达,质荷比为3940和3400的蛋白质峰在散发性肠腺瘤患者中高表达.以质倚比分别为5640、3160和4290的蛋白质峰为基础,联合质荷比为3940、13 750和4300的蛋白质峰所建立的模型判别效果最佳,对FAP与散发性肠腺瘤的判别准确率分别为94.7%和93.7%.结论 SELDI-TOF-MS能有效筛选FAP与散发性肠腺瘤的差异表达蛋白,支持向量机方法所建立的质谱模型判别效果较好,为进一步研究FAP的分子发病机制提供了切入点.     

NGAL基因表达上调与大肠肿瘤发生的关系研究

目的:研究大肠腺瘤-腺癌序列过程中中性粒细胞明胶酶相关脂质运载蛋白(NGAL)和基质金属蛋白酶9(MMP-9)表达的变化及相互关系,以阐明其在大肠癌发生发展中的作用.方法:采用组织芯片与免疫组织化学SP法检测94例瘤旁/癌旁组织、92例腺瘤(其中管状腺瘤36例,管状绒毛状腺瘤47例,绒毛状腺瘤9例)、54例腺瘤伴不典型增生或癌变及60例腺癌中NGAL和MMP-9蛋白的表达情况.结果:在大肠腺癌、腺瘤伴不典型增生或癌变、腺瘤、瘤旁/癌旁组织中,NGAL蛋白表达的阳性率分别为55%(33/60)、37%(20/54)、23%(21/92)和2%(2/94),MMP-9蛋白表达的阳性率分别为74%(44/60)、44%(24/54)、25%(23/92)、6%(6/94),NGAL和MMP-9蛋白的表达在腺瘤、腺瘤伴不典型增生或癌变肠腺癌组织中显著高于瘤旁/癌旁组织,差异具有统计学意义(P<0.01),并且在正常肠黏膜→腺瘤→腺瘤伴不典型增生或癌变→腺癌这一过程中两者的表达逐渐增高,差异具有统计学意义(P<0.01),然而二者在腺瘤组中管状腺瘤、管状绒毛状腺瘤、绒毛状腺瘤三者之间的表达差异无统计学意义(P>0.05).NGAL与MMP-9蛋白的表达在整个序列过程中(正常肠黏膜组、腺瘤组、腺瘤伴不典型增生或癌变组、腺癌组)呈显著正相关(rs=0.622,P<0.01).结论:NGAL蛋白和MMP-9蛋白表达的上调可能在大肠腺瘤-腺癌序列的发展过程中发挥一定作用,二者的异常表达可能参与大肠腺癌的发生.     

直肠癌组织Akt-mTOR蛋白表达与临床病理因素相关性分析

目的:探讨直肠癌组织Akt-mTOR蛋白表达与临床病理特征和预后的相关性。方法:应用免疫组织化学技术检测Akt和mTOR蛋白在60例直肠癌组织、30例直肠腺瘤组织及10例癌旁正常黏膜组织中的表达,并对60例患者进行随访。结果:Akt蛋白在直肠癌组织、直肠腺瘤及正常直肠黏膜组织中的阳性表达率分别为60.0%(36/60)、33.3%(10/30)和10.0%(1/10),其在直肠癌组织与直肠腺瘤组织、正常直肠黏膜组织中的表达差异均有统计学意义(χ2值分别为5.692和8.600,P值分别为0.017和0.003),直肠腺瘤组织与正常直肠黏膜组织中的表达差异无统计学意义,χ2=2.048,P=0.152;mTOR蛋白在直肠癌组织、直肠腺瘤及正常直肠黏膜组织中的阳性表达率分别为61.7%(37/60)、36.7%(11/30)和10.0%(1/10);其在直肠癌组织与直肠腺瘤组织、正常直肠黏膜组织中的表达差异均有统计学意义(χ2值分别为5.022和9.220,P值分别为0.017和0.002),直肠腺瘤组织与正常直肠黏膜组织中的表达差异无统计学意义,χ2=2.540,P=0.111。两者在直肠癌组织中过表达并成正相关,r=0.583,P<0.05。Akt蛋白表达水平与患者的术前CEA水平、TNM分期、淋巴结转移和远处转移有关,P<0.05;与患者的年龄、性别、肿瘤部位、形态和分化程度无关,P>0.05;mTOR蛋白表达水平与患者的术前CEA水平、分化程度、TNM分期、淋巴结转移和远处转移有关,P值均<0.01;与患者的年龄、性别、肿瘤部位和形态无关,P>0.05。Cox回归模型分析显示,远处转移是直肠癌的独立预后危险的因素。结论:Akt-mTOR信号通路在直肠癌组织中表达水平均与CEA水平、TNM分期、淋巴结转移和远处转移有关,但其与预后的关系仍需进一步研究。     

FAP-1与大肠癌的实验研究

目的 通过观察大肠癌中Fas相关磷酸酯酶(FAP-1)的表达,了解肿瘤细胞增殖和凋亡状态,探讨FAP-1表达与细胞增殖和凋亡的关系,分析其临床意义.方法 采用免疫组化SP法检测50例大肠癌及20例大肠腺瘤组织标本中FAP-1蛋白的表达水平,10例正常肠黏膜组织为对照组.结果 FAP-1蛋白的阳性表达主要定位在细胞浆内,少数也可见于核周.FAP-1蛋白在正常肠黏膜组织中呈阴性表达,大肠腺瘤中的表达率为25.0%(5/20),大肠癌中的阳性表达率76.0%(38/50),与前两者相比差异有显著性(P＜0.01),而对照组与大肠腺瘤组差异无显著性(P＞0.05).FAP-1的表达与大肠癌的临床Dukes分期、病理分级相关,而与年龄、性别、肿块大小、转移性无明显相关性.结论 FAP-1在大肠癌组织中有广泛表达,提示FAP-1可能对于大肠癌的发生、发展具有一定意义.     

结肠癌组织FADD和IPO-38蛋白表达与预后相关性分析

目的 结肠癌的发生发展过程与细胞过度增生和细胞凋亡受抑制密切相关,而凋亡促进蛋白FADD与增殖相关蛋白IPO-38分别具有调控细胞凋亡和显示细胞增殖能力的作用,本研究旨在研究结肠癌组织FADD和IPO-38蛋白的表达情况,探讨二者在结肠癌发生发展中的作用及其与预后的关系.方法 采用免疫组化方法对FADD和IPO-38蛋白在50例正常结肠黏膜、50例结肠腺瘤和280例结肠腺癌样本中的表达情况进行检测.结果 FADD蛋白在正常结肠黏膜、结肠腺瘤、无淋巴结转移结肠腺癌及有淋巴结转移结肠腺癌中表达阳性率分别为100.00％、98.00％、79.83％和72.04％,其蛋白表达呈逐渐降低趋势,H=52.515,P＜0.001;FADD蛋白表达与结肠癌有无淋巴结转移之间差异有统计学意义,Z=-2.705,P=0.007.IPO-38蛋白在正常结肠黏膜、结肠腺瘤、无淋巴结转移结肠腺癌及有淋巴结转移结肠腺癌的阳性表达率分别为82.00％、100.00％、100.00％和100.00％,其蛋白表达等级逐渐增强,H=232.987,P＜0.001;IPO-38蛋白表达与结肠癌有无淋巴结转移之间差异有统计学意义,Z=-3.072,P=0.002.在结肠腺癌中,FADD和IPO-38蛋白的表达呈负相关,rls=-0.263,P＜0.001.FADD蛋白低表达是影响结肠癌患者预后的独立因素.结论 FADD和IPO-38蛋白的表达在结肠癌的发生发展过程中起一定的作用.FADD低表达可作为结肠癌预后不良的一个参考指标.     

Receipt of colorectal testing prior to colorectal carcinoma diagnosis

BACKGROUND: Among patients with established colorectal carcinoma, the prior use of procedures for colorectal carcinoma screening is unknown. In addition, the association of disease stage at the time of diagnosis with previous procedure use has not been studied previously at a population level. METHODS: The Surveillance, Epidemiology, and End Results tumor registry files identified 5806 patients age >or=70 years with an initial colorectal carcinoma diagnosis in 1999. Medicare claims data from 1995 through diagnosis were extracted and identified receipt of fecal occult blood testing (FOBT), sigmoidoscopy, colonoscopy, and barium enema. Time intervals were divided into >6 months prior to diagnosis and or=1 procedures >6 months before diagnosis, with FOBT the most frequent procedure (36%). Colonoscopy was performed in only 6% of patients prior to the 6-month peridiagnostic period. Compared with patients who underwent no procedures or who underwent procedures only within 6 months of diagnosis, those who underwent procedures >6 months earlier presented with earlier stages of carcinoma. CONCLUSIONS: The receipt of colorectal procedures in the interval >6 months prior to a diagnosis of colorectal carcinoma was low, suggesting the under use of screening in this population. The association of procedure use with earlier stage at presentation as well as unmeasured benefits in reducing carcinoma incidence suggest beneficial effects of this testing.     

Metastatic colorectal cancer

Opinion statement Despite advances in screening procedures and the use of adjuvant therapy, approximately 50% of patients with colorectal cancer eventually will develop metastatic disease. Long-term disease-free survival can be achieved in 25% to 40% of selected patients who undergo resection of liver or lung metastases. For all other patients, treatment is palliative. For decades, 5-fluorouracil was the only available drug for colorectal cancer; hence, numerous trials were performed that used various administration schedules and modulating agents to improve therapeutic efficacy. The addition of leucovorin to 5-FU improves response but not survival. Infusion schedules alter the toxicity profile but have a negligible impact on survival. Irinotecan was the first new drug to demonstrate activity in colorectal cancer. It was used initially in the second-line setting, where it was shown to improve quality of life and survival over best supportive care or infusional 5-FU. Recently, irinotecan has been incorporated into the front-line treatment of metastatic colorectal cancer in combination with 5-FU and leucovorin; this combination improves survival by approximately 3 months. Careful patient selection and adherence to strict dose adjustments are essential to prevent significant toxicity when patients are treated on this regimen. The oral fluoropyrimidine capecitabine recently has been approved for the front-line treatment of patients with colorectal cancer who are not appropriate candidates for combination therapy. Oxaliplatin, a novel DACH (diaminocyclohexane) platinum with definite activity in colorectal cancer, is approved for this disease in Europe and is undergoing phase III clinical trials in the United States. Other drugs with potential activity in colorectal cancer include raltitrexed, pemetrexed disodium, and the epothilone analog BMS-247550 (Bristol-Myers Squibb, New York, NY). Novel cytostatics with promising activity in colorectal cancer are being evaluated in clinical trials, including epidermal growth factor receptor inhibitors, such as IMC-C225 (Imclone Systems, New York, NY) and ZD1839 (AstraZeneca, London, UK), angiogenesis inhibitors such as bevacizumab and SU5416 (Sugen, San Francisco, CA), and vaccines such as CEAVac (Titan Pharmaceuticals, San Francisco, CA). For those patients whose disease is localized to the liver, there also is an emerging role for local therapies, including cryosurgery, radiofrequency ablation, and hepatic artery infusional chemotherapy, and resection. The emergence of these new drugs and new interventional modalities has allowed physicians who treat colorectal cancer to move beyond 5-FU.     

Primary colorectal lymphoma

This study aimed at investigating the incidence, presentation, patient and tumor characteristics, treatment, and outcome of primary colorectal lymphomas (PCL) at a tertiary care center in Lebanon over a 25-year period. The Dawson’s criteria were used for selection of eligible cases. The medical records were reviewed for demographic variables, the presence of risk factors, presenting signs and symptoms, method of diagnosis, histologic type, type of therapy, and condition at last follow-up. Nine cases of PCL were identified (12.7% of gastrointestinal lymphomas and 0.1% of colorectal malignancies). The mean age at presentation was 44.2 years with male predominance noted. Abdominal pain was the most common presentation (77.8%). Colonoscopy was performed for eight patients with non-specific gross tumor characteristics. Three patients had Burkitt’s and six had diffuse large B-Cell lymphomas. The most common site of involvement was the cecum (55.6%) with all cases presenting in stage I_E. Surgery was performed for six patients followed by chemotherapy except for one, and three patients had chemotherapy only. The median survival time was 25 months and the 2-year survival time was approximated at 60%. It is concluded that PCL is a rare malignancy with well-identified disease characteristics yet controversial ideal management plan.     

Metastatic colorectal cancer

Metastatic colorectal cancer remains a public-health issue on a global scale. With development of a new generation of cytotoxic agents, survival has improved for patients with metastatic disease. How to maximize the benefit of chemotherapy with acceptable toxicity remains incompletely answered. Hepatic resection can provide a significant hope for long term survival, and a subset of patients might benefit from perioperative approaches. More recently, specific molecular processes have been targeted for therapeutic interventions, and encouraging results have been achieved using inhibitors of the Epidermal Growth Factor Receptor and the Vascular Endothelial Growth Factor.     

结直肠相关多原发癌临床特点分析

目的研究结直肠相关多原发癌（MPC）的临床特点,并与同期单发结直肠癌（CRC）比较分析,以为早期诊断及治疗提供帮助。方法回顾分析1997年9月至2007年11月收治的病理资料及分期明确的573例CRC及MPC患者的相关临床资料,比较中位生存期（mOS）、肿瘤病理分型、分期、发病部位等。结果573例患者中MPC45例（7．85％）,其中有肠外病灶的MPC好发部位依次是胃、乳腺、卵巢、肺、小肠及其他。肠内多原发结直肠癌（MPCC）病例中,发病部位以升结肠癌最多（34．0％）,而CRC则以直肠癌最多（36．5％）。比较单发CRC与MPC,两者在肿瘤家族史方面无明显差异;中位发病年龄分别为57岁和63岁;有结肠息肉史的MPC患者占20．0％,而单发CRC仅0．9％;单发CRC与MPC的mOS分别为93．7和64．8个月。单发CRC和MPC病理类型都以高一中分化腺癌居多,但MPC中黏液腺癌较多。结论结直肠相关MPC在CRC患者中较常见;MPC特别是MPCC中结直肠息肉更多见;结直肠相关MPC比单发CRC的mOS短,提示结直肠相关MPC的预后可能较单发CRC差;MPCC结直肠息肉多发,肿瘤发病间隔时间短,mOS也短,提示预后较有肠外肿瘤的结直肠相关MPC差。