Radiation-induced morphological changes and radiocurability in squamous cell carcinoma of the head and neck region. A preliminary report.

Tissue samples taken from 22 patients before and during radical irradiation of squamous cell carcinomas in the head and neck region were studied by light and electron microscopy. The changes in keratinization pattern at the ultrastructural level seemed to be correlated with the outcome of the radiotherapy. The irradiation induced several cellular changes, of which nuclear atypia was the most prominent. This atypia was considered to be mainly due to cell death rather than to an aggressive nature of the tumor, because the number of mitoses decreased at the same time. The tumor invasion pattern remained unchanged. The keratinization pattern remained almost unchanged at the light microscopical level, but a slight increase of intracellular filaments and desmosomes was found in the electron microscopic study. The amount of intercellular filaments increased in three patients out of four with complete remission (CR), but in no case with tumor dissemination (n = 3) during radiotherapy. In patients with local persistent tumor or a local recurrence (LP + LR) (n = 15) the filaments either increased, decreased or remained unchanged. The number of desmosomes either increased or remained unchanged in three of four CR patients, in 13 of 15 LP + LR patients and in only one of three patients with tumor dissemination. They decreased in two patients with tumor dissemination, but only in one case with CR and in 2 cases with LP + LR. It is suggested that changes in cytoskeleton and desmosomes might be important in anchorage of tumor cells locally and might have value for prediction of the tumor response to radiotherapy. Further studies on larger materials are, however, needed before more definite conclusions can be drawn.     

Eccrine sweat gland tumor of clear cell origin involving the eyelids.

A 47-year-old patient with an unusual tumor involving the right upper and lower eyelids has been followed for almost 6 years. The tumor has remained localized to the eyelids and has recurred locally following attempts at complete or partial excision. The morphological features of the tumor as seen by ordinary light microscopic methods were puzzling, and resulted in a variety of pathologic diagnoses. Light microscopic examination of plastic-embedded semithin sections, and electron microscopic examination indicate that this is a hitherto undescribed eccrine sweat gland tumor of clear cell origin. Its infiltrative growth pattern and tendency to local recurrence suggests that it may be a low-grade malignant neoplasm.     

Desmoplastic malignant melanoma: A light and electron microscopic study of two cases.

This light and electron microscopic study of two recent cases of desmoplastic malignant melanoma (DMM) attempts to resolve the conflict in views regarding the nature of the cells responsible for the desmoplasia associated with this clinicopathologic entity. On the basis of evidence presented, it is concluded that the cells are dedifferentiated tumor cells with fibroblastic features and probably functions, rather than host engendered fibroblasts in response to invasive melanoma. The evidence includes: observation of macular desmosomes between tumor cells, an unheralded feature previously noted in amelanotic and melanotic melanomas; electron microscopic observation of fibroblast-like cells by others in spindle cell squamous carcinomas; and light microscopic features of malignancy including vascular invasion in one of the two cases. A reproducible light microscopic pattern diagnosis of this variant of malignant melanoma is reaffirmed in both cases.     

Tumor radiosensitization by antiinflammatory drugs: evidence for a new mechanism involving the oxygen effect

We hypothesized that nonsteroidal antiinflammatory drugs (NSAIDs) might enhance tumor radiosensitivity by increasing tumor oxygenation (pO2), via either a decrease in the recruitment of macrophages or from inhibition of mitochondrial respiration. The effect of four NSAIDs (diclofenac, indomethacin, piroxicam, and NS-398) on pO2 was studied in murine TLT liver tumors and FSaII fibrosarcomas. At the time of maximum pO2 (t(max), 30 minutes after administration), perfusion, oxygen consumption, and radiation sensitivity were studied. Local pO2 measurements were done using electron paramagnetic resonance. Tumor perfusion and permeability measurements were assessed by dynamic contrast-enhanced magnetic resonance imaging. The oxygen consumption rate of tumor cells after in vivo NSAID administration was measured using high-frequency electron paramagnetic resonance. Tumor-infiltrating macrophage localization was done with immunohistochemistry using CD11b antibody. All the NSAIDs tested caused a rapid increase in pO2. At t(max), tumor perfusion decreased, indicating that the increase in pO2 was not caused by an increase in oxygen supply. Also at t(max), global oxygen consumption decreased but the amount of tumor-infiltrating macrophages remained unchanged. Our study strongly indicates that the oxygen effect caused by NSAIDs is primarily mediated by an effect on mitochondrial respiration. When irradiation (18 Gy) was applied at t(max), the tumor radiosensitivity was enhanced (regrowth delay increased by a factor of 1.7). These results show the potential utility of an acute administration of NSAIDs for radiosensitizing tumors, and shed new light on the mechanisms of NSAID radiosensitization. These results also provide a new rationale for the treatment schedule when combining NSAIDs and radiotherapy.     

Characterization of early kidney lesions in estrogen-induced tumors in the Syrian hamster

Syrian hamsters were treated with diethylstilbestrol (DES), a potent estrogen and kidney carcinogen, or ethinyl estradiol (EE), a strong estrogen but weak carcinogen, for 1-9 months. At monthly intervals their kidneys were studied using light, immunoperoxidase, and electron microscopic techniques. At 5 months, DES-treated animals exhibited interstitial lesions composed of small round cells with a high nuclear:cytoplasmic ratio. Immunoperoxidase and ultrastructural studies showed these cells to be similar to cells in fully formed tumors at 9 months. Early lesions in EE-treated animals (seen as early as 1 month) were dissimilar, these lesions appeared in the deep cortex adjacent to the renal pelvis, where proximal tubules underwent hyperplastic changes, showing columnar cells with large nuclei, occasional mitoses, and sloughing of apical cytoplasm. Cells in early lesions of EE-treated animals did not resemble the fully developed tumor in either immunoperoxidase or ultrastructural features; although with longer treatment these tubular lesions progressed to dysplasia (3-5 months) and severe dysplasia/carcinoma in situ (7 months), they did not form grossly visible tumors during the 9-month study. Both early lesions identified were specific, inasmuch as they were not observed in control animals and animals treated with beta-dienestrol and 17 alpha-estradiol (noncarcinogenic weak estrogens). Animals given a combination of DES and EE showed tubular hyperplasia but not interstitial lesions; this finding was of particular interest because hamsters given this combination of estrogens do not develop gross renal tumors. These results strongly implicate the primitive interstitial cell in the hamster kidney as the cell of origin of the DES-induced neoplasm.     

Ultrastructural change due to acquired cisplatin resistance in human bladder cancer cells

Cis-diaminedichloroplatinum (cisplatin) has therapeutic efficacy against advanced bladder cancer. However, the response is often limited due to appearance of drug-resistant tumor cells. Recently, we established a cisplatin-resistant human bladder cancer cell line, T24/R2, from T24 human bladder cancer cell line, which shows 18-fold resistance to cisplatin by the stepwise exposure of increasing concentrations of cisplatin. A light microscopic (LM) and transmission electron microscopic (EM) examination was performed to investigate the morphological and ultrastructural changes during induction of drug resistance. In LM, the cytoplasm of T24R2 cells showed plumper pattern than that of T24 parent cells. In EM, the chromatin pattern of T24R2 cells was finely dispersed compared to T24 cells, which were coarse and aggregated. The mitochondrial volume, rough endoplasmic reticulum, polyribosomes and ribosomes were moderately increased in T24R2 cells. The cell membrane showed ruffled border and great amount of double membrane vesicles and pinocytic vesicles were observed in the cell surface of T24R2 cells, which were seldom observed in T24 cells. With these findings, we concluded that human bladder cancer cells underwent morphological and ultrastructural changes during acquiring of resistance to cisplatin. We could suggest that these changes might be involved in the drug resistance mechanism in human bladder cancer cells.     

The in vitro effect of the tumor promoter 12-O-tetradecanoylphorbol-13-acetate on Sertoli cell morphology.

The objective of the present study was to examine the effects of the well-known tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) on the morphology of cultured Sertoli cells from immature rats. The cells were cultured for 5 days and the TPA was added at the end of the culture period for 8 h at a concentration of 10-7 M. Viability tests showed that controls as well as TPA-treated cells remained viable during the culture period and no deleterious effects were observed as a result. Application of computerized morphometry at both light and electron microscopic levels revealed that TPA caused important changes in cell morphology in vitro. Statistical analysis of the results indicated that compared to the controls, Sertoli cells treated with TPA exhibited fewer astrocytic-type cytoplasmic extensions and a smaller size. Our results support the conclusion that the tumor promoter TPA, when applied to immature Sertoli cells in vitro, causes significant morphological alterations.     

三氧化二砷诱导人鼻咽低分化鳞癌裸鼠移植瘤细胞分化的研究

背景与目的:既往研究发现三氧化二砷(As2O3)可诱导白血病细胞的分化,但对实体瘤细胞的诱导分化作用报道甚少。本实验拟探讨As2O3对人鼻咽低分化鳞癌可移植瘤在BALB/C裸鼠体内生长的影响,重点观察其诱导鼻咽癌细胞分化的作用。方法:以人鼻咽低分化鳞癌鼠移植癌细胞株CSNE-1为研究对象,观察腹腔注射As2O3(5mg·kg-1·d-1连续10天后,每周给药3次,连续3周)对人鼻咽癌移植瘤在BALB/C裸鼠体内生长情况的影响。用光学显微镜和电子显微镜观察移植瘤细胞形态变化,用免疫组化法检测瘤组织中增殖细胞核抗原(PCNA)的表达情况。结果:腹腔注射As2O3后,人鼻咽癌BALB/C裸鼠移植瘤生长被抑制,抑瘤率为75.4%。同时,瘤组织中癌细胞密度减少,细胞皱缩,胞浆红染,瘤组织分化渐成熟,出现角化细胞和角化珠;间质结缔组织增多。透射电镜下肿瘤细胞出现成熟分化及明显角质化,细胞表面微小突起增多,细胞间桥粒增多并以桥粒互相连接,细胞核浆比例减少,胞浆中出现大量张力原纤维并围绕核周。癌细胞PCNA在阴性对照组呈高表达,PI(PCNA阳性细胞指数)为(95.2±5.0)%,而As2O3组癌细胞PCNA表达明显减少,PI为(53.6±7.0)%(P<0.001)。结论:As2O3抑制人鼻咽低分化鳞癌BALB/C裸鼠移植瘤的生长,这种抑制作用可能与其诱导癌细胞向成熟方向分化有关     

UVB-irradiation regulates VLA-4-mediated melanoma cell adhesion to endothelial VCAM-1 under flow conditions

The major aspect contributing to the mortality of melanoma is its ability to spread, or metastasize. Ultraviolet B light (UVB) is considered an indirect cause of melanoma formation. However, little is known about the potential effects of UVB to melanoma metastasis. Integrins, a large family of cell adhesion molecules (CAMs) expressed on the melanoma cell surface, are important for cell signaling, growth, and migration during metastasis. Most critically, tumor cell tissue invasion is dependent on the initial interaction of tumor cells with vascular endothelium at the target organ, and there is increasing evidence for a prominent role of melanoma very late antigen‐4 (VLA‐4) integrin binding to its endothelial ligand vascular cell adhesion molecule‐1 (VCAM‐1) in this process. This research focuses on the quantitative modulation of VLA‐4 integrin expression and function on melanoma cells after UVB irradiation. The present data show that at 3, 12, and 18 h post‐UVB irradiation, VLA‐4 expression was unchanged relative to untreated cells, but adhesion to VCAM‐1 decreased significantly. Immunofluorescence studies implied that the spatial organization of VLA‐4 on the melanoma cell surface contributed to the changes in avidity for VCAM‐1 upon UVB irradiation. With increased understanding of the molecular mechanisms underlying melanoma–endothelial interactions upon UVB irradiation, clinical advances for melanoma may be developed. © 2010 Wiley‐Liss, Inc.     

T-lymphocyte mediated tumor cell destructionin vivo associating with a specific feature of apoptosis

The immunological responsiveness of the T-cell immunodificient NC nude mice tumor models was reconstructed by thymic transplantation of the semidominance NC mice. After immune reconstruction (IR) the tumor continued to grow until 2 to 3 weeks, then the volume of the tumor reduced gradually and disappeared at the 9th week. In both H901 and SW1116 solid tumor nodules was found by light microscopic study (IM), after IR, tumor cells gradually replaced by lymphocytes and fibroblasts, shrinked till only isolated cell groups, then totally disappeared. The whole processes like that the tumor cells were nibbled. It was found that the main tumor cell death related with a specific feature of apoptosis, which had typical dense chromatin distributed along the inner surface of the nuclear membrane by transmission electron microscopic study (TEM). The IR model could be useful for further mechanical research of immune system.     

Ultrastructural and immunohistochemical study of an adult case of chordoid meningioma

A 64-year-old male patient presented with generalized convulsions. Magnetic resonance imaging revealed a large meningeal tumor with some cysts in the right frontal region. Surgical resections were performed three times, and local radiation therapy was administered twice over a period of 8 years for the treatment of tumor recurrences. The tumor tended to recur in spite of the surgical and radiation therapies. The tumor was diagnosed as a chordoid meningioma, and the second surgical specimen showed increasing nuclear atypia and mitoses in tumor cells. An immunohistochemical study revealed the tumor cells were positive for vimentin, S-100 protein, and cytokeratin AE1/AE3. An electron microscopic study revealed intracytoplasmic vacuolar spaces, loosely connected interdigitating cell processes with intermediate junctions, and extracellular spaces which contained fluffy granular intercellular substances. The tumor cell surfaces displayed pseudopodia which extended into the intercellular spaces and the tumor cells had moderate quantities of cytoplasm containing abundant mitochondria and glycogen granules. According to the ultrastructural features in the past reports of chordoid meningiomas, these meningiomas are suspected to have a mixture of the characteristic ultrastructural features of meningothelial meningiomas and chordoid sarcomas. This case was presented at the 16th annual meeting of the Japan Society of Brain Tumor Pathology in Fukuoka, Japan, on May 28, 1998.     

Balloon cell malignant melanoma of the skin. A clinicopathologic study of 34 cases with histochemical, immunohistochemical, and ultrastructural observations.

Balloon cell malignant melanoma (BCMM) is a rare histologic variant of malignant melanoma (MM). Thirty-four patients with BCMM from the files of the Armed Forces Institute of Pathology (AFIP) were studied by means of clinicopathologic correlation and histochemical, immunohistochemical, and ultrastructural methods to better define this entity. The cytoplasmic features of the balloon cells observed in BCMM resemble those noticed in balloon cell nevus (BCN), but the presence of nuclear pleomorphism, atypia, and mitoses and the absence of intervening stroma help distinguish BCMM. The cells also show many histochemical, immunochemical, and ultrastructural features of conventional melanoma cells. Although it is generally believed that balloon melanoma cells represent a degenerative change, the immunohistochemical and electron microscopic findings suggest that the balloon tumor cells are most likely metabolically active melanocytic cells. Microscopically, BCMM also must be differentiated from other clear cell tumors such as clear cell sarcoma (MM of soft parts), hibernoma, xanthoma, sebaceous neoplasms, metastatic renal cell carcinoma, (malignant) clear cell acrospiroma, (malignant) granular cell tumor, granular (clear) cell basal cell carcinoma, clear cell syringoma, and atypical fibroxanthoma. The prognosis of BCMM usually correlates with the tumor thickness similar to that in other histologic types of cutaneous MM. Nineteen (57.5%) of 33 patients with adequate follow-up information died of disseminated tumors from 2 months to 12 years after the initial treatment. Six (18.2%) patients developed local recurrences: four of these patients died of metastasis and two were alive with metastatic tumor at last contact. Five (15.2%) patients were alive with metastatic tumors, and seven (21.2%) were alive without evidence of disease at last contact. Recognition of BCMM is important because of its malignant biologic behavior.     

Differential modulation of nuclear texture, histone acetylation, and MDR1 gene expression in human drug-sensitive and -resistant OV1 cell lines

Image cytometric study of pathological specimens or cell lines has suggested that epigenetic mechanisms are likely to play a major role in determining chromatin patterns evaluable through nuclear texture analysis. We previously reported that nuclear textural changes observed in the OV1-VCR etoposide-resistant ovarian carcinoma cell line were associated with an increased acetylated histone H4 level. In this study we analyzed the effects of treatments with the HDAC inhibitor trichostatin A (TSA) or with nickel subsulfide on histone H4 acetylation, nuclear texture, and MDR1 gene expression in drug-sensitive IGROV1 and drug-resistant OV1-VCR cell lines. In IGROV1 cells, TSA induced an increase in acetylated H4 level associated with a chromatin textural decondensation and an increase in MDR1 gene expression. In OV1-VCR cells, a similar increase in H4 acetylation was observed, but nuclear texture or MDR1 gene expression remained unchanged. ChIP analysis revealed that MDR1 gene expression remained stable in TSA-treated OV1-VCR cells despite a localized increase in H4 acetylation at the promoter level. Analysis of the methylation status of MDR1 promoter showed an increase in DNA methylation at 3 specific sites in OV1-VCR cells, that could participate to TSA low responsiveness in these cells. Treatment with nickel subsulfide induced a decrease in H4 acetylation without any effect on nuclear texture characteristics in both cell lines. In OV1-VCR cells, nickel subsulfide induced a significant down-regulation of the MDR1 gene expression. These results indicate that modulation of histone H4 acetylation level can be associated with up- or down-regulation of the MDR1 gene in OV1 cells. However, this modulation does not always result in chromatin pattern alterations and these data emphasize the complexity of chromatin texture regulation in tumor cells.     

T－LYMPHOCYTE MEDIATED TUMOR CELL DESTRUCTIONIN VIVO ASSOCIATING WITH A SPECIFICFEATURE OF APOPTOSIS

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Malignant mesenchymoma. Case report with electron microscopic study.

A malignant tumor from the thigh of a 65-year-old man was examined by light and electron microscopy. The tumor was diagnosed by light microscopy as malignant mesenchymoma with prevalence of leiomyosarcomatous or liposarcomatous components, but electron microscopic study revealed that the basic cell was similar to chondroblastic or osteoblastic cells. However, a precise ultrastructural classification of the tumor cells was not possible, since the majority of the cells had mixed features. The histogenesis, differentiation, and relation to prognosis are discussed.     

Effects of quinolinone derivative, vesnarinone, in combination with irradiation on human lung cancer cell lines

The quinolinone derivative, vesnarinone, is a novel inotropic agent used in the treatment of heart failure. It has also been found that vesnarinone has a potential anti-cancer activity. To evaluate the anti-cancer activity of vesnarinone in combination with irradiation, we investigated the cytostatic and cytotoxic effects on human lung cancer cell lines (PC-9 and Lu 134A) using MTT assay and isobologram analysis. We also analyzed the nuclear fragmentation of tumor cells by flow cytometric analysis. Our study demonstrated that combination of vesnarinone with irradiation had an additive inhibitory effect on both PC-9 and Lu 134A tumor cell growth. Vesnarinone could improve the sensitivity of tumor cells to irradiation and thus the dose of irradiation could be reduced to half without decreased inhibitory effect. Significant increase of the tumor cell nuclear fragmentation was observed with combination of vesnarinone and irradiation. These results indicate that vesnarinone not only directly inhibited tumor cell growth, but also improved the sensitivity to irradiation. Combination of vesnarinone with irradiation may be an efficacious protocol for lung cancer treatment. The inhibitory effect was ascribed to inducing tumor cell apoptosis.     

Bone marrow cytogenetics: the lineage of dividing cells changes during the first few hours in culture

To determine changes in the cell lineages of metaphases karyotyped following different culture times, marrow from 11 healthy individuals was studied using a technique that allows simultaneous analysis of karyotype and cell lineage. Cell lineage was identified as erythroid by surface glycophorin A, granulocytic by Sudan black B and PM-81, and monocytic by lysozyme. Marrow examined sequentially showed granulocytic mitoses to initially decrease from a mean of 40% at 1.75 hr to 6% at 3.5 hr and then increase, being 46% by 6 hr and 82% after 1 day, and remain high for the 10 days studied. Erythroid mitoses were most frequent (mean, 72%) at 3.5 hr and then decreased rapidly, being 16% by 6 hr, 7% at 1 day, and absent thereafter. When granulocytic mitoses were least frequent, 20-36% of mitoses were also unreactive with glycophorin A. Double staining experiments to identify these cells found some to be monocytic, but most remained unidentified. The authors conclude that mitoses of different hematopoietic lineages predominate when normal marrow is studied cytogenetically at different times following aspiration, and that the major changes occur during the first 8 hours. These findings have importance for how cytogenetic studies are performed in leukemia.     

高强度聚焦超声体外破坏原发性肝癌的病理学观察

目的 探讨高强度聚焦超声（high-intensity focused ultrasound,HIFU)体外破坏原发性肝癌（HCC）的病理学变化。方法 HIFU治疗56例HCC患者,6例患者治疗后外科手术切除肿瘤,观察其病理学变化。结果 在光镜下,HIFU治疗区和非治疗区边界清楚,治疗区内肿瘤细胞核出现固缩、碎裂或溶解等不可逆性细胞损伤现象,血窦结构塌陷,内皮细胞受损严重,靶区边缘有新生肉芽组织形成,逐渐修复坏死组织,在电镜下,肿瘤细胞、细胞器和细胞核出现明显损伤,细胞膜、核膜连续性丧失,核碎裂,细胞浆内出现大小不等的小泡样结构。结论 HIFU体外治疗HCC是安全、有效和可行的。     

Histological changes after single high-dose irradiation for squamous cell carcinoma arising from a burn scar

Single high-dose irradiation has become a safe and encouraging form of radiation therapy but variations in the histological changes after irradiation have not been fully understood in clinical settings. The aim of this study was to investigate the histological changes of tumor cells after single high-dose, direct irradiation with a case of cutaneous squamous cell carcinoma that developed on a burn scar. The patient received irradiation followed by radical resection of the tumor. Tissues before and after irradiation were stained with Hematoxylin and Eosin (H&E), TdT-mediated dUTP-X nick end labeling (TUNEL), and by immunohistochemistry for MIB-1, p53, and p21. Massive necrosis was noted after irradiation but a few viable tumor cells remained. Preradiation histology revealed MIB-1 and p53 as positive, and p21 tumor cells as partially positive. These expressions were reduced after irradiation. The TUNEL stain was consistently negative. It was established that tumor cell death after single high-dose irradiation was caused mainly by necrosis rather than apoptosis in clinical settings.