胰腺癌血清相关蛋白质分子的差异表达

目的 利用双向差异凝胶电泳(2D-DIGE)的方法建立胰腺癌、慢性胰腺炎和正常对照人群外周血清的差异蛋白质双向凝胶电泳图谱并分析差异蛋白质点.方法 双向差异凝胶电泳分离20例胰腺癌患者、10例慢性胰腺炎患者和20例正常对照组人群外周血清蛋白,比较不同血清中蛋白质表达的差异.结果 成功建立胰腺癌、慢性胰腺炎和正常人之间的外周血清蛋白质双向差异凝胶电泳图谱,软件分析共发现了168个明显差异表达的蛋白质点,其中在胰腺癌组与正常对照组的比较中有22个蛋白质点在胰腺癌血清中表达上调,29个蛋白质点下调;在胰腺癌组和慢性胰腺炎组的比较中有24个蛋白质点在胰腺癌血清中表达上调,54个表达下调;在慢性胰腺炎组和正常对照组的比较中有20个蛋白质点表达上调,19个蛋白质点表达下调.结论 双向差异凝胶电泳是快速有效的分离蛋白质新的方法,得到的双向差异凝胶电泳图谱以及显著表达差异的蛋白质点为质谱鉴定提供了实验基础.

Abstract：

Objective To analyze and compare the proteomes expressed by human pancreatic carcinoma, chronic pancreatitis and normal control groups. Methods Differencial expression of the proteomes in peripheral serum was analyzed by fluorescence 2-D difference gel electrophoresis (2D-DIGE). Results 2D-DIGE protein maps in 20 patients with pancreatic cancer, 10 patients with chronic pancreatitis and 20 normal controls were analyzed and 168 spots were identified by gel-analysis software. Between pancreatic cancer group and normal control group 22 protein spots were up-regulated and 29 spots down-regulated; 24 spots were up-regulated and 54 spots down-expressed between pancreatic cancer group and chronic pancreatitis group; 20 spots were up-regulated and 19 spots down-regulated between chronic pancreatitis group and normal control group. Conclusion 2D-DIGE was a rapid and efficient method to separate proteins. 2D-DIGE protein maps and different protein spots would provide an exprimental basis for the phase of mass spectrometry.     

Profile of neuroblastoma detected by mass screening, resected after observation without treatment: results of the Wait and See pilot study

Background/Purpose

Neuroblastoma (NB) detected by mass screening (MS) usually shows favorable prognosis and sometimes regresses spontaneously. Therefore, the authors started an observation program for these patients to avoid overtreatment. In this study, the authors analyzed the profile of NB resected after observation to elucidate the nature of NB detected by MS.

Methods

Between 1994 and 2004, 22 NB patients matched the following criteria and entered the observation program after obtaining informed consent: stage I or II, less than 5 cm in diameter, and without involvement of large vessels or organs. If increase in size, elevation of tumor markers, or evidence of metastasis was observed, the tumor was immediately resected.

Results

Thirteen (59%) of 22 cases showed spontaneous regression. In the remaining 9 cases, tumors were resected because of parents' request, increase in size, and/or elevation of tumor markers. Four tumors had at least one unfavorable biologic feature, and 3 of them had more than 2. According to Shimada's system, 2 had unfavorable histology. One was diploid tumor, 3 had 1p deletion, and Trk-A expression was low in 4 tumors. All patients survived without evidence of recurrence.

Conclusions

The observation program has shown that at least one third of the NB detected by MS regressed spontaneously. On the other hand, MS may detect some cases with unfavorable tumor in early stage, which benefit from screening.     

Profiling of nuclear extract proteins from human neuroblastoma cell lines: the search for fingerprints

Purpose

Neuroblastoma (NB) commonly presents with advanced disease at diagnosis and is associated with poor survival. If identified early, however, survival is improved suggesting a benefit of early detection. The authors have used proteomics technology in an attempt to identify novel markers that permit early detection of NB and characterize its molecular makeup.

Methods

Three different human NB cell lines SK-N-AS, SK-N-DZ, and SK-N-FI were subjected to series of biochemical fractionation steps to extract nuclear proteins. These proteins were analyzed for differential expression by 2-dimensional polyacrylamide gel electrophoresis. Polypeptides of interest were subsequently identified by liquid chromatography-linked tandem mass spectrometry.

Results

Multiple proteins were identified in these human NB cell lines including SET (a ubiquitous nuclear protein), stathmin (a cytosolic signal transduction protein), and grp94 (a heat shock protein). SET is a putative oncogene associated with the chromosomal translocation (6;9) leading to acute undifferentiated leukemia. Stathmin is an oncogene found in greater abundance in leukemic cells compared to nonleukemic cells. A total of 94-kDa glucose-regulated protein has been shown to be protective in human breast cancer cells in vitro and related with the occurrence, differentiation, and progression of human lung cancer. The first protein has not been previously associated with NB.

Conclusions

The identification of these 3 previously unrecognized cancer-related potential biomarkers in human NB cell lines may prove useful in developing diagnostic tests. The proteomic methodology of 2-dimensional polyacrylamide gel electrophoresis/mass spectrometry also provides an improved opportunity to understand the natural history of NB and develop novel chemotherapeutic agents for this prevalent childhood malignancy with a dismal outcome.     

大鼠颈7神经根切断后肱三头肌蛋白质组学研究

目的 用荧光差异双向凝胶电泳技术(2-D DIGE)分析大鼠颈7(C7)神经根切断后肱三头肌蛋白质表达变化规律,初步探讨其功能代偿的分子机制.方法 建立一侧C7神经根切断大鼠模型,对侧为对照组,分别在C7神经根切断后第1、2、3、6、12周取等量的肱三头肌,用荧光差异双向凝胶电泳技术分离、单因素方差分析(one-way ANOVA)筛选蛋白质表达量差异具有统计学意义的蛋白质点(P＜0.05),再对差异蛋白质点进行胶内酶解,并用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)或串联质谱技术进行鉴定.结果 在大鼠C7神经根切断前和切断后的不同时间点,肱三头肌有38个蛋白质点表达发生了明显变化,其中有27个蛋白质被确切地鉴定,有11个蛋白质与肌纤维的能量代谢相关.在C7切断后第1周,无氧代谢酶表达下调,第3周无氧代谢酶、氧化酶(有氧代谢酶)表达均上调,但在第6～ 12周发生代谢类型的转变,无氧酶上调、氧化酶下调,同时线粒体蛋白质翻译延长因子表达下调.结论 C7神经根切断后肱三头肌功能代偿机制可能与肌纤维类型的转变有关,而这种转变的调节可能发生在能量代谢酶的翻译水平.     

The role of surgery in localized neuroblastoma

A 30-year experience with 83 patients, median age two years, with Children's Cancer Study Group (CCSG) Stages I, II, and III localized neuroblastoma was studied to determine factors that influence outcome. In addition, histology was reclassified in all patients based on the Shimada system, which is divided into five subtypes according to age and cytohistologic criteria. A multivariant survival analysis was carried out and patients were considered to have failed if they relapsed or died from any cause. Initial analysis determined that CCSG stage, Shimada histologic classification, and presence of disease in lymph nodes were statistically significant predictors of failure. Histology was the most important factor with Shimada subtypes 1, 2, and 4 having good outcome and 3 and 5 poor outcome. The latter three variables were combined to create four prognostic groups that had distinctly different rates of survival. Further analysis showed that after controlling for prognostic groups, extent of surgery was a statistically significant predictor. Patients who had more complete surgical resection had better disease-free survival.     

Immunohistochemical expressions of Cap43 and Mina53 proteins in neuroblastoma

Background

We studied the expressions of both Mina53, which is a myc target gene and is related to cell proliferation, and Cap43, which is related to metastasis suppression and downregulation of MYCN gene, in neuroblastoma.

Methods

Forty-eight surgically obtained neuroblastoma specimens were immunohistochemically stained. The Cap43 and Mina53 expression levels were determined, and their relationship to clinical prognostic factors, biological prognostic factors, and the patients' prognosis were examined.

Results

The Cap43 expression score was significantly high in the cases that had one of the good prognostic factors (<1 year old, early stage, mass screening case, no MYCN gene amplification), whereas the Mina53 expression score was high in those with poor prognostic factors. Regarding the MYCN expression site, the Cap43 expression score was significantly high in the cases demonstrating cytoplasm expression, whereas the Mina53 expression score was significantly high in the cases demonstrating nucleus expression. A significant relationship was found between Cap43 and TrkA, between Mina53 and Ki-67, and between Mina53 and TrkA. The prognosis was significantly favorable in the Cap43 high-expression cases, whereas it was significantly poor in the Mina53 high-expression cases.

Conclusions

Cap43 and Mina53 are both considered to be important biological and prognostic factors in neuroblastoma.     

Long-term observations in cases with spontaneous carotid-cavernous fistulas

Summary Twenty-six cases with spontaneous carotid-cavernous fistulas were followed up for periods ranging between 4 months and 9 years 8 months. A complete regression of symptoms without reappearance for more than 6 months was noted in 19 cases, a marked improvement in 2 cases, and a moderate regression in 3 cases. In 2 cases, symptoms have continued for 9 years 8 months and for 1 year. The regression of symptoms was usually delayed in patients less than 60 years old, in cases in which the symptoms developed slowly, and in cases with multiple draining veins. According to our observations a regression of symptoms may occur after very slight changes of haemodynamics.Compression of the cervical carotid artery for a short time or a temporary occlusion of the carotid artery by a balloon catheter should be considered as the treatment of choice in the first instance in cases with spontaneous carotid-cavernous fistulas showing relatively low pressure and low flow shunt.     

The role of multimodal therapy in patients with local and regional neuroblastoma

The records of 49 patients with local and regional neuroblastoma treated at The Children's Hospital of Philadelphia between 1972-1981 were reviewed to determine the contribution of radiation therapy and chemotherapy to their management. All 11 state I patients were treated with surgery alone and 10 (91%) survived for periods ranging from 3 to 10 years. Half of the 24 stage II patients received radiation therapy with or without chemotherapy and 8/12 (67%) survive. Ten of the 12 remaining stage II patients survive (83%) following surgery alone. All stage II patients had residual disease and 13 had involvement in lymph nodes. 7 in the combined treatment group and 6 in the surgery alone group. All 14 stage III patients received postoperative chemotherapy and radiation therapy (RT) and 7 or 50% remain disease free. We conclude that surgery alone is sufficient for patients with stage I NBL, that RT and chemotherapy do not appear to alter the outcome in stage II patients, and better methods of therapy are needed for patients with stage III disease.     

Thymoma with spontaneous regression and disappearance of pleural effusion

A 31-year-old woman was admitted to our hospital with sudden onset of chest pain. Chest radiography and computed tomography (CT) on admission showed an anterior mediastinal tumor with left pleural effusion, which was diagnosed as an inoperable malignant mediastinal tumor. However, 3 weeks after admission CT showed that the tumor was diminishing and the pleural effusion had disappeared without any treatment. CT-guided needle biopsy was performed, but diagnosis was impossible because most of the specimen was necrotic. A biopsy during video-assisted thoracic surgery was then performed. The intraoperative finding showed that the tumor was round, well mobilized, and did not invade adjacent structures. It was then assumed to be a benign teratoma that had been ruptured into the thoracic cavity. The operation was converted to a thoracotomy to resect it, but it could not be completely resected because of inflammatory adhesions to the mediastinum. Two months later, total thymectomy was performed through a median sternotomy because the tumor was pathologically diagnosed as a thymoma.     

Massive adrenal hemorrhage in neonatal neuroblastoma

In a neonate with exsanguinating intraperitoneal bleeding admitted with a provisional diagnosis of ruptured liver due to birth trauma, laparotomy revealed the source of hemorrhage to be an adrenal neuroblastoma. This case prompted a review of cases of abdominal neuroblastoma admitted to the Neonatal Surgical Unit in Alder Hey Children's Hospital from 1953 to 1976. The features of 10 cases are presented: in three of them there was hemorrhage into the tumor. Of the 10 cases, six survived tumor free from 2 to 12 yr and there is one short-term survivor. The purpose of this presentation is to emphasize the possibility of an underlying neuroblastoma in cases of neonatal adrenal hemorrhage and also the relatively good prognosis in neuroblastoma presenting in the neonatal period.     

Experimental study on the growth and proliferation kinetics of residual tumor after surgery in mouse neuroblastoma

In order to elucidate the proliferation kinetics of residual tumors after surgery, experimental studies were performed, using C-1300NB and A/J mice, tumor-host system. Mice were inoculated with C-1300NB cells in the chest and leg simultaneously, and then divided into three groups. Growth curves of chest tumors (residual tumors) in Group B after amputation of the tumor-bearing leg were significantly steeper than those of both Group A, whose tumor-bearing legs were not amputated, and Group C, whose normal legs were amputated, at the same tumor age. 3H-TdR labeling indices of chest tumors of Group B were significantly higher than those of Group A (P less than 0.05). DNA histograms of the chest tumors of Group A uniformly showed a unimodal distribution with a peak in the 2c range. On the other hand, in Group B the peak of nuclear DNA distribution shifted from the 2c range to the 4c range after amputation of tumor-bearing leg. The results indicated that an increase in cells in the DNA synthetic phase occurred in the remaining neuroblastoma after reducing the volume of tumor in host by amputation of tumor-bearing legs.     

Hedgehog signaling pathway in neuroblastoma differentiation

Purpose

The hedgehog (Hh) signaling pathway is activated in some adult cancers. On the other hand, the Hh signaling pathway plays an important role in the development of the neural crest in embryos. The aim of this study is to show the activation of Hh signaling pathway in neuroblastoma (NB), a pediatric malignancy arising from neural crest cells, and to reveal the meaning of the Hh signaling pathway in NB development.

Methods

This study analyzed the expression of Sonic hedgehog (Shh), GLI1, and Patched 1 (Ptch1), transactivators of Hh signaling pathway, by immunohistochemistry in 82 NB and 10 ganglioneuroblastoma cases. All 92 cases were evaluated for the status of MYCN amplification.

Results

Of the 92 cases, 67 (73%) were positive for Shh, 62 cases (67%) were positive for GLI1, and 73 cases (79%) were positive for Ptch1. Only 2 (10%) of the 20 cases with MYCN amplification were positive for Shh and GLI1, and 4 cases (20%) were positive for Ptch1 (MYCN amplification vs no MYCN amplification, P ≦ .01). The percentage of GLI1-positive cells in the cases with INSS stage 1 without MYCN amplification was significantly higher than that with INSS stage 4. Of 72 cases without MYCN amplification, 60 were GLI1-positive. Twelve cases were GLI1-negative, and the prognosis of the GLI1-positive cases was significantly better than that of the GLI1-negative cases (P = .015).

Conclusions

Most of NBs without MYCN amplification were positive for Shh, GLI1, and Ptch1. In the cases without MYCN amplification, the high expression of GLI1 was significantly associated with early clinical stage and a good prognosis of the patients. In contrast to adult cancers, the activation of the Hh signaling pathway in NB may be associated with the differentiation of the NB.     

Evaluation of the role of surgery in 130 patients with neuroblastoma

The role of surgery for children with neuroblastoma was evaluated by using a recently proposed TNM staging system. One-hundred thirty patients were retrospectively assigned a TNM clinical stage (CS) preoperatively and a pathologic stage (PS) postoperatively. Patients with CS 4 were separated into CS 4A and CS 4B according to their age and pattern of metastases. Patient survival was analyzed according to CS, age, location of primary, and PS. Actuarial survival of patients was as follows: CS 1, 100%; CS 2, 82%; CS 3, 63%; CS 4A, 50%; and CS 4B, 5%. For all stages, patients younger than 1 year old survived longer than those older than 1 year (72% v 32%). Prognosis for CS 1 was the same regardless of age. For CS 2 and CS 3, patients younger than 1 year old lived longer. CS 4A had better survival than CS 4B. Survival by site was 100% for cervical, 62% for mediastinal, 45% for pelvic, and 36% for retroperitoneal primaries. The role of surgery was evaluated by analyzing survival according to the postoperative PS. PS 1-2-3 A were regarded as satisfactory resections since all macroscopic tumor was removed. PS 3B as a debulking procedure, and PS 3C as an unresectable lesion which was biopsied. Patients with nonmetastatic disease (CS 1-3) with PS 1 and PS 2 disease had a 100% survival rate; PS 3A, 93%; PS 3B, 58%; and PS 3C, 21%. This proves the value of total resection in nonmetastatic disease. The role of surgery could also be proven in metastatic disease.(ABSTRACT TRUNCATED AT 250 WORDS)     

神经母细胞瘤Survivin基因表达和细胞凋亡的研究

目的：探讨神经母细胞瘤（NB）Survivin表达和细胞凋亡。方法：应用免疫组织化学技术和DNA原位末端标记技术检测26例NB中Survivin基因表达和细胞凋亡。结果：NB中15例有Survivin表达阳性细胞检出，阳性检出率57．7％。神经节母细胞瘤（GNB）的阳性检出率为20．0％，NB为66．7％；预后差组织（UFH）组为88．9％，预后好（FH）组这41．2％；I－Ⅱ及ⅣS期为27．3％，Ⅲ－Ⅳ期为80．0％。NB组、UFH组、Ⅲ-Ⅳ期标本Survivin阳性表达率高于GNB、FH、I－Ⅱ及ⅣS期标本，P值分别为0．0823、0．0243、0．0105。17例有凋亡细胞检出NB中12例有Survivin阳性表达，Survivin标记指数与凋亡指数（AI）呈负相关（n=12,γ=-0.591）、与生存时间呈负相关（n=12,γ=-0.629），AI与生存时间则呈正相关（n=16,γ=0.563）。结论：Survivin引起的凋亡抑制与NB发生发展关系密切，是患者预后的重要指标。     

超声诊断儿童颈部神经母细胞瘤

目的探讨儿童颈部原发性神经母细胞瘤(NB)的超声表现及其诊断价值。方法分析154例临床可疑儿童颈部原发NB的超声图像。采用χ~2检验比较超声诊断结果与病理学诊断结果的差异,以病理结果作为金标准,计算超声诊断儿童颈部原发NB的敏感度、特异度、阳性预测值及阴性预测值。结果超声诊断原发颈部NB结果与病理学结果的差异无统计学意义(χ~2=92.827,P0.05)。超声诊断NB的敏感度为95.45%(21/22),特异度为93.94%(124/132),阳性预测值为72.41%(21/29),阴性预测值为99.20%(124/125)。儿童颈部原发性NB典型超声表现:颈正中线一侧的边界清晰的实性占位、无液化(21/21,100%);瘤体内部以低回声为主(19/21,90.48%),伴细碎小钙化灶(20/21,95.24%)。结论儿童颈部NB的超声表现具有一定的特异性。超声检查可作为诊断颈部NB的首选影像学检查。     

Late complications after surgery in patients with neuroblastoma

Purpose

The purpose of this study was 2-fold: to study the complications of neuroblastoma treatment and to establish surgical guidelines.

Materials and Methods

The medical records of 92 patients with neuroblastoma (stage 1 or 2, n = 33; stage 3, n = 21; stage 4, n = 31; stage 4S, n = 7) who had undergone surgery in our department between 1985 and 2001 were reviewed, with an emphasis on treatments and late complications.

Results

The disease-free survival rate was 98.3% (58/59) in infantile patients, whereas it was 36.4% (12/33) in advanced patients older than 1 year. The rate was improved up to greater than 50% after the introduction of intensive local therapy with intraoperative radiation (IOR). The treatment-associated morbidity rate was 15.0% in nonadvanced infantile patients, was 42.1% in advanced infantile patients, and was 33.3% in advanced older patients. In the advanced cases, renovascular problems were most frequently seen especially after IOR. Furthermore, pulmonary problems, cardiac problems, and second cancer were highlighted as fatal problems; thyroid dysfunction, vertebral deformity, and growth retardation were also noted.

Conclusions

Higher incidences of late morbidity and treatment-associated mortality appeared to be more strongly related to disease stage rather than to patient age. Appropriate surgery and IOR with lower doses should be recommended as a surgical strategy for high-risk patients.     

Challenges of laparoscopic resection of abdominal neuroblastoma with lymphadenectomy

BACKGROUND: The laparoscopic procedure involving total resection of abdominal neuroblastoma combined with lymphadenectomy has not been reviewed in English literature. The aim of this study was to evaluate the significance and accuracy of laparoscopic resection of abdominal neuroblastoma. METHODS: Since July 1997, five patients with abdominal neuroblastoma underwent laparoscopic resection combined with lymphadenectomy or sampling of the lymph nodes. The length of operation, intraoperative blood loss, resectability, and complications were retrospectively reviewed and evaluated. RESULTS: Four cases were managed laparoscopically, but one case was converted to open procedure because of poor visualization around large vessels. The mean operation time was 135 min and the intraoperative blood loss 52 ml. CONCLUSIONS: Good visualization of the primary tumor and large vessels is, arguably, the most important factor for successful completion of this procedure laparoscopically. Precise indicators for laparoscopic resection of abdominal neuroblastoma provide a better prognosis and a good quality of life for children with neuroblastoma.