The distribution of CD34-positive stromal cells and myofibroblasts in colorectal carcinoid tumors

In order to understand the stromal reaction associated with colorectal neoplasms, we examined specimens from 26 patients including normal colorectal tissues (n=15), carcinoid tumors (n=12), well differentiated adenocarcinomas (n=10), and poorly differentiated adenocarcinomas (n=4), using an immunohistochemical method. Myofibroblasts and CD34-positive stromal cells were distributed in the mucosa and in the area between the submucosal and subserosal layers, respectively. However, the distribution of these cells markedly changed with the invasion of neoplasms. Namely, myofibroblasts were abundant in the invasive stroma of all colorectal neoplasms. CD34-positive stromal cells were completely absent from the invasive stroma of colorectal cancers. On the other hand, CD34-positive stromal cells were absent from four out of five carcinoid tumor cases with lesions measuring less than 2 mm in size, but were present in all seven cases of carcinoid tumors measuring more than 2 mm. Double-immunostaining identified stromal cells expressing both ASMA and CD34 in several carcinoid tumor cases. Finally, no CD34-positive stromal cells were observed in the invasive stroma of colorectal cancers. However, the distribution of these cells in carcinoid tumors may depend on the lesion size. Namely, CD34-positive stromal cells existed between neoplastic nests in large-sized carcinoid tumors. Myofibroblasts in the stroma of colorectal neoplasms may originate from CD34-positive stromal cells.     

Immunological evaluation of personalized peptide vaccination in combination with UFT and UZEL for metastatic colorectal carcinoma patients

To investigate the safety and immunological responses of personalized peptide vaccination in combination with oral administration of UFT and UZEL for metastatic colorectal carcinoma (mCRC), fourteen patients were enrolled in the present study. Peptides were determined based on the presence of peptide-specific cytotoxic T lymphocyte precursors and IgG in each patient. A maximum of four peptides were subcutaneously administered weekly with UFT (300 mg/m² day⁻¹) and UZEL (75 mg/day) for 4 weeks, followed by 1 week of rest. This therapy was well-tolerated although there was a grade-3 skin reaction at the vaccination site in one patient. An increase in peptide-specific interferon-γ production or peptide-specific IgG after the tenth vaccination was observed in nine of ten or eight of ten patients tested, respectively. IgG responses were well correlated with overall survival (P = 0.0215). The safety and immunological responsiveness of the present therapy suggest that this combination would be of clinical benefit for mCRC patients, and further trials are merited. T. Hattori and T. Mine equally contributed to this work.     

ATF2与ATF3在结直肠癌中表达及临床病理意义

目的：观察结直肠癌组织中活化转录因子2（ATF2）和活化转录因子3（ATF3）的表达并分析其表达的临床病理意义。方法：收集结直肠癌病例,明确其病理诊断并收集临床资料,应用免疫组织化学SP法检测活化转录因子2和活化转录因子3蛋白的表达。结果：ATF2在癌旁肠组织、腺瘤、腺癌组的阳性表达率分别为38%,32%,64%,差异有统计学意义,其中癌旁肠组织、腺瘤分别与腺癌有显著性差异。ATF3在癌旁肠组织、腺瘤、腺癌组的阳性表达率分别为56%,44%,52%,差异无统计学意义。ATF2表达与浸润肠壁深度,淋巴结转移有关,而与肿块大小、部位、分化程度无关。ATF3表达与肿块直径、浸润肠壁深度,而与淋巴结转移、部位、分化程度无关。结论：ATF2与结直肠癌反生和发展有关,而ATF3与结直肠癌的恶性演进有关。     

ATF2 与ATF3 在结直肠癌中表达及临床病理意义

目的:观察结直肠癌组织中活化转录因子2(ATF2)和活化转录因子3(ATF3)的表达并分析其表达的临床病理意义。方法:收集结直肠癌病例,明确其病理诊断并收集临床资料,应用免疫组织化学SP法检测活化转录因子2和活化转录因子3蛋白的表达。结果:ATF2在癌旁肠组织、腺瘤、腺癌组的阳性表达率分别为38%,32%,64%,差异有统计学意义,其中癌旁肠组织、腺瘤分别与腺癌有显著性差异。ATF3在癌旁肠组织、腺瘤、腺癌组的阳性表达率分别为56%,44%,52%,差异无统计学意义。ATF2表达与浸润肠壁深度,淋巴结转移有关,而与肿块大小、部位、分化程度无关。ATF3表达与肿块直径、浸润肠壁深度,而与淋巴结转移、部位、分化程度无关。结论:ATF2与结直肠癌反生和发展有关,而ATF3与结直肠癌的恶性演进有关。     

ICAM-1和CD44v6在结、直肠癌中的表达及临床意义

目的:了解ICAM-1和CD44v6在结、直肠癌中的表达,探讨它们与结直肠癌侵袭与转移的关系.方法:应用免疫组织化学pv-9000二步法检测ICAM-1和CD44v6在40例结直肠癌和30例大肠良性腺瘤组织中的表达情况,并分析其与结直肠癌临床病理特征的关系.结果:40例结、直肠癌和30例大肠良性腺瘤中,ICAM-1的表达阳性率分别是72.5%(29/40)、33.3%(10/30),CD44v6的表达阳性率分别是88.9%(27/40)、26.7%(8/30). ICAM-1和CD44v6的表达与年龄、性别、肿瘤细胞分化程度无关,但与淋巴结转移、Dukes分期具有相关性(p<0.05).有淋巴结转移、Dukes分期在C+D期者,ICAM-1和CD44v6的表达阳性率均明显升高.ICAM-1和CD44v6在大肠良性腺瘤中呈低表达或不表达.ICAM-1和CD44v6两者均阳性表达时,其预测结、直肠癌淋巴结转移的特异性升高,有一定的临床意义.结论:CD44v6和ICAM-1的阳性表达预示结直肠癌具有较强的侵袭转移能力,可作为预测结直肠癌转移潜能的生物学指标.     

Quantitative biophysical metrics for rapid evaluation of ovarian cancer metastatic potential

Ovarian cancer is routinely diagnosed long after the disease has metastasized through the fibrous submesothelium. Despite extensive research in the field linking ovarian cancer progression to increasingly poor prognosis, there are currently no validated cellular markers or hallmarks of ovarian cancer that can predict metastatic potential. To discern disease progression across a syngeneic mouse ovarian cancer progression model, here we fabricated extracellular matrix mimicking suspended fiber networks: cross-hatches of mismatch diameters for studying protrusion dynamics, aligned same diameter networks of varying interfiber spacing for studying migration, and aligned nanonets for measuring cell forces. We found that migration correlated with disease while a force-disease biphasic relationship exhibited F-actin stress fiber network dependence. However, unique to suspended fibers, coiling occurring at the tips of protrusions and not the length or breadth of protrusions displayed the strongest correlation with metastatic potential. To confirm that our findings were more broadly applicable beyond the mouse model, we repeated our studies in human ovarian cancer cell lines and found that the biophysical trends were consistent with our mouse model results. Altogether, we report complementary high throughput and high content biophysical metrics capable of identifying ovarian cancer metastatic potential on a timescale of hours.     

Quantitative evaluation of immunohistochemical staining in gastrointestinal stromal tumors

OBJECTIVE: To evaluate the differences among pathologists' interpretations in gastrointestinal stromal tumors (GISTs) and to demonstrate the usefulness of quantitative pathology in the assessment of immunohistochemical staining. STUDY DESIGN: Twenty GISTs were separately evaluated by 4 pathologists by the visual estimation method using a 6-antibody panel. Each case was then quantitatively measured with a computer-assisted image analysis system by 2 pathologists. Cohen's kappa test was performed for statistical analysis. RESULTS: All GISTs showed some degree of expression of CD 117, CD34, SMA and Ki-67. No case was immunoreactive for desmin or S-100 protein. There were remarkable differences in the pathologists' visual estimations. Moreover, the discrepancies between visual and quantitative methods were noteworthy. The differences in interpretations showed the greatest variability for Ki-67, which is known to be related to poor prognosis. CONCLUSION: Quantitative pathology in assessment of immunohistochemical staining of GISTs may improve the consistency in the interpretation of staining results and provide some degree of reproducibility.     

Cleavage of carcinoembryonic antigen induces metastatic potential in colorectal carcinoma

Carcinoembryonic antigen (CEA), a widely used tumor marker, is attached by a glycosylphosphatidylinositol (GPI) anchor motif to the cell membrane. Recent study suggested that membrane-bound CEA might be cleaved by glycosylphosphatidylinositol-phospholipase D (GPI-PLD). We studied the effect of GPI-PLD on the cleavage of CEA to elucidate the implication for metastatic potential in colorectal carcinoma cells. CEA amount of conditioned medium was changed by suramin and phenanthroline (activator and inhibitor of GPI-PLD) only in SW620 and SW837 which expressed both CEA and GPI-PLD mRNA. Suramin treatment also augmented migratory activity and decreased cell surface CEA expression in SW620 and SW837. Furthermore, GPI-PLD knockdown cells using GPI-PLD-specific siRNA in SW620 and SW837 showed decreased CEA secretion from cell membrane and the migration activity, increased membrane-bound CEA amount. Splenic injection of SW620 and SW837 induced marked hepatic metastases in nude mice. These results suggest that membrane-bound CEA is cleaved by GPI-PLD and that this cleavage enhances the metastatic potential in colorectal carcinoma cells.     

The distribution of myofibroblasts and CD34-positive stromal cells in normal renal pelvis and ureter and their cancers

In this article, we examined the distribution of myofibroblasts and CD34-positive stromal cells in normal renal pelvis and ureter and their cancers using immunohistochemistry. Eighteen tumors and normal tissues apart from the main tumor were examined. In the wall of normal renal pelvis and ureter, no myofibroblasts were observed through all layers, but CD34-positive stromal cells were observed in the deep area of lamina propria, muscular layer and adventitia. In the stroma of renal pelvic and ureteral cancers, myofibroblasts were distributed in fifteen tumors and were absent in three tumors. All three tumors containing no myofibroblasts in the stroma were non-invasive type and all invasive cancers contained myofibroblasts in the stroma. CD34-positive stromal cells were consistently absent in the stroma of cancers, irrespective of the invasiveness. Finally, myofibroblasts are major stromal components in renal pelvic and ureteral cancers, particularly in invasive cancers, and CD34-positive stromal cells are consistently absent or lost in the stroma of their cancers. These findings suggest that the invasion of renal pelvic and ureteral cancers may cause the phenotypic change of stromal cells.     

Negative CEA values in metastatic colorectal carcinoma and the likelihood of complete chemotherapy response

INTRODUCTION: Experimental results reported in the literature have suggested that CEA might inhibit host defense mechanisms and that immunotolerance to CEA could play an important role in the development of metastases in colorectal carcinoma. It might therefore be assumed that negative CEA values during metastatic disease represent a favorable prognostic factor. Surprisingly, there are very few data available about negative CEA. The aim of this study was to determine the significance of negative initial CEA values in patients with metastatic colorectal carcinoma. PATIENTS AND METHODS: Initial CEA values were determined in 114 patients with metastatic colorectal carcinoma. The patients were divided into three groups according to these values: I (n=22) <5 ng/mL; II (n=33) 5-100 ng/mL; III (n=59) >100 ng/mL. RESULTS: Seven/114 complete responses (CR), 22/114 partial responses (PR), 45/114 instances of stable disease (SD) and 38/114 of progressive disease (PD) were registered, while two patients were not evaluable. There were six long-lasting CRs (median 24 months, range 10-37 months) in the CEA-negative patient subset, while in the CEA-positive subset there was only one CR, in a patient with an initial CEA level of 18 ng/mL. The mean initial CEA values in the different response categories were: CR: 4.0 ng/mL; PR: 436 ng/mL; SD: 1442 ng/mL; PD: 6071 ng/mL. The likelihood of response, in particular CR, was highly dependent upon CEA levels (Fisher's exact test, 0.00001). The median survival decreased significantly with increased values of CEA (p=0.006). CONCLUSION: Negative CEA in metastatic disease was the main characteristic of the patient subset capable of attaining CR. When relapsing, all patients but one became CEA positive.     

Quantitative analysis of caveolin-rich lipid raft proteins from primary and metastatic colorectal cancer clones

Caveolin-rich lipid rafts (CLRs) are thickened sections of the cell membrane that are composed of the integral membrane proteins caveolins together with saturated long chain fatty acids, cholesterol and lipids. Membrane proteins - lipid raft proteins in particular - may play important roles in cell signaling and cell-cell interaction. Due to their unique structure, CLRs seem to be the preferred docking site for specific proteins involved in focal adhesion and cancer metastasis. Our objective was thus to identify and quantify CLR proteins from primary and metastatic colorectal cancer (CRC) clones. We found differential expression of nine CLR proteins from primary and metastatic CRC clones. Among the identified proteins, an immune system inhibiting protein was significantly overexpressed in the metastatic clone, while cell adhesion and transport molecules were among the overexpressed proteins in the primary clone. All the identified CRL proteins are involved in tumorigenesis, specifically metastasis, and may thus serve as therapeutic targets. A novel concept for identification and quantification of CLR proteins with label-free mass spectrometry method was specifically examined in this study. Validation of the method against immunoblotting and FACS analysis indicates that it can be applied for the identification of novel biomarkers for cancer and metastasis.     

Appearance and distribution of stromal myofibroblasts and tenascin-C in feline mammary tumors

Myofibroblasts and extracellular matrix protein tenascin-C (Tn-C) are known to be implicated in cancer progression in human cancer. In feline mammary tumors that are a suitable model for human breast cancer, however, little is known about stromal myofibroblasts and no information is available on the expression of Tn-C. Feline samples of normal mammary glands and proliferating mammary lesions were routinely processed and serial sections were cut and immunostained with anti-α-smooth muscle actin (α-SMA) or Tn-C antibody. Myofibroblasts were not included in the stroma of 90% (9/10) of normal mammary gland tissues, 92% (12/13) of adenosis, and 63% (5/8) of simple adenomas. On the other hand, all 40 simple carcinomas contained stromal myofibroblasts to a varied extent. Tn-C expression was detected in the stroma of 92% (37/40) of carcinomas, and its global distribution almost coincided with that of myofibroblasts. In addition, Tn-C immunoreactivity was occasionally observed in the basement membrane zone around ducts in some cases of normal mammary glands and benign lesions, but barely observed in the stroma. These results suggest that stromal myofibroblasts may be a major cellular source of Tn-C and be involved in malignant progression of feline mammary tumor.     

The distribution pattern of myofibroblasts in the stroma of human bladder carcinoma depends on their invasiveness

The presence of myofibroblasts has been elucidated in the stroma of neoplasm of various organs. In the present article, we studied the distribution of myofibroblasts in the stroma of bladder carcinoma. Twenty-five surgical resected bladder tumors (urothelial carcinoma, n = 21; combined urothelial carcinoma and adenocarcinoma, n = 2; sarcomatoid squamous cell carcinoma, n = 1; combined urothelial carcinoma and squamous cell carcinoma, n = 1) were selected and we evaluated the distribution of myofibroblasts using immunohistochemical, electron and immunoelectron microscopic techniques. Immunohistochemically, the distribution pattern of myofibroblasts in invasive and non-invasive carcinomas were predominantly fascicular and reticular forms, respectively. Moreover, myofibroblasts around bladder carcinoma cells were confirmed by electron microscope. Understanding the distribution pattern of myofibroblasts in the stroma of bladder carcinoma may provide available information about the presence of carcinoma invasion.     

p53 increases experimental metastatic capacity of murine carcinoma cells.

Transfection of a cloned p53 gene into a murine bladder carcinoma cell with a low metastatic capacity led to elevated levels of p53 protein in clonal transfectants. After intravenous inoculation into syngeneic mice, p53-transfected clones showed significantly increased metastatic potential in comparison with control transfectants. The observed change did not seem to be due to a change in growth potential per se since the cell lines showed similar growth properties in vitro.     

分化抑制因子-1在大肠癌中的表达及临床意义

目的:研究分化抑制因子-1(Id-1)在人大肠癌组织中的表达及其与临床病理特征的关系。方法:应用免疫组织化学方法(SP法)检测Id-1在56例大肠癌组织及56例远癌肠黏膜中的表达水平,并分析其与临床病理特征的关系。结果:在大肠癌组织中Id-1的过表达率为80.4%,在远癌肠粘膜中过表达率为12.5%,两组比较差异有统计学意义(P0.01)。Id-1过表达与大肠癌Dukes分期及淋巴结转移有关(P0.05),而与患者年龄、性别、肿瘤组织分化程度无关(P0.05)。结论:Id-1过表达可能参与大肠癌演进,Id-1可作为判断大肠癌恶性程度和预后的指标。     

Quantitative evaluation of the uptake capacity of intact thrombocytes using fluorescent dyes

A new approach to quantitative determination of fluorescent dye uptake by intact cells is suggested. Fluorescent amine acridine orange selectively accumulating in 5HT granules of platelets has been used. Fluorescence signal analysis allows the estimation of a relative granule volume and the ratios of acridine orange transfer over cytoplasmic and granule membranes. The following results were obtained in human and rabbit platelets: a relative granule size was 14 +/- 1 % and 29 +/- 2 % of the total cell volume, intra-granule to extra-granule dye concentration ratios were 2260 +/- 382 and 30000 +/- 5550, while intra-cytoplasm to extra-cytoplasm concentration ratios were 375 +/- 60 and 225 +/- 60, respectively.     

Human adipose tissue–derived mesenchymal stromal cells and their phagocytic capacity

Mesenchymal stromal cells (MSCs) have evidenced considerable therapeutic potential in numerous clinical fields, especially in tissue regeneration. The immunological characteristics of this cell population include the expression of Toll‐like receptors and mannose receptors, among others. The study objective was to determine whether MSCs have phagocytic capacity against different target particles. We isolated and characterized three human adipose tissue MSC (HAT‐MSC) lines from three patients and analysed their phagocytic capacity by flow cytometry, using fluorescent latex beads, and by transmission electron microscopy, using Escherichia coli, Staphylococcus aureus and Candida albicans as biological materials and latex beads as non‐biological material. The results demonstrate that HAT‐MSCs can phagocyte particles of different nature and size. The percentage of phagocytic cells ranged between 33.8% and 56.2% (mean of 44.37% ± 11.253) according to the cell line, and a high phagocytic index was observed. The high phagocytic capacity observed in MSCs, which have known regenerative potential, may offer an advance in the approach to certain local and systemic infections.