Human ovarian mucinous cystadenocarcinoma cell line (MN-1) was established and then cultured 113 times during a period of about 20 months. Furthermore by single cell cloning of the MN-1 cell line, two different morphological and cytological cell lines (MN-1 A, MN-1 B) have been established. MN-1 cells were found to form a jig-saw puzzle, pavement-like pattern showing no contact inhibition. It was observed that the cells had variable shapes; especially conspicuous among these were a small round type and an elongated spindle type. The population doubling time of MN-1 cells was about 29.4 hours at the 47th passage of culture. Karyotype analysis of the MN-1 cells showed that the number of chromosomes was hypertetraploidic: the modal number being 102. Following heterotransplantation of MN-1 cells who BALB/C nude mice, the tumor developed and came to have cystic and solid components, which were similar to those of the original tumor.
MN-1 A cells showed a pavement-like pattern, with individual cells being small and round in shape. Their doubling time was about 40.0 hours at the 22nd passage of culture. According to our analysis, a hypotriploid tendency was observed in the chromosomal number. MN-1 B cells, on the other hand, had an elongated spindle shape, showing a jig-saw puzzle pattern. Their doubling time was about 32.0 hours at the 9th passage of culture, and the number of chromosomes showed a hypertetraploid tendency. After MN-1 A and MN 1 B cells had been separately transplanted into nude mice, the MN-1 B cell tumor was observed to develop more rapidly than that of the MN-1 A cells. The tumor figure for the MN 1 A consisted mainly of cystic components, which were similar to well differentiated mucinous cystadenocarcionma. On the other hand, the tumor figure for the MN-1 B line was composed of solid parts analogous to poorly differentiated mucinous cystadenocarcinoma.
We have now established a human ovarian mucinous cystadenocarcinoma cell line (MN-1), and separated two distinct morphological cytological cell lines, i.e., MN-1 A and MN-1 B. As a result, it has been demonstrated that MN-1 is made up of more than one clone.

View full abstract

Papanicolaou stained uterine cervical smears containing the “ground glass” appearance of nuclet with multinucleated giant cells suggestive of herpes simplex virus infection were destained and reprocessed for in situ hybridization using a biotinylated probe for herpes simplex virus DNA.
Six slides were processed in this way.
A hybridization signal for viral DNA was noted in the nuclet of cells having the “ground glass appearance”. This simple and rapid non-radioactive detection system is effective in uterine cervical smears prepared as much as 3 years previously, indicating that this technique is suitable for retrospective studies.

View full abstract

Cytology is a useful technique in the diagnosis of cervical cancer. It is still difficult, however, to make a correct cytopathological diagnosis of early stage Ib cancer (cases with invasion of less than 5mm excluding stage Ia as defined by the criteria of the Japan Society of Obstetrics and Gynecology).
The cytological specimens of 13 cases with stage Ia and 22 cases with early stage Ib (6 cases with invasion less than 3mm=Ib 1, 16 cases with invasion up to 5mm=Ib 2) were analyzed to clarify differences among the various stages. The incidences of multinuclear, malignant pair cell and nucleolus were similar among the 3 groups. Anisonucleosis, elongated nucleus and differentiative tendencies for cellular figures were prominent in cases with Ib 2. No tumor diathesis was observed in 11 of 13 cases with stage Ia (84.6%), whereas 10 of 16 cases (62.5%) with early stage Ib had apparent diathesis. Cellular clusters were not in evidence in 10 cases of Ia (69.2%) nor in 2 of Ib 2 (12.5%), whereas numbers of clusters exceeding 10 were observed in 0.0% and 25.0%, respectively. The cluster size tended to be large in Ib 2 and cluster diameters exceeding 0.3mm were found in 23.1% of Ia and 62.5% of Ib 2. We conclude that prominent anisonucleosis, elongated nucleus, differentiative tendencies and large clusters, over 0.3mm in diameter, numerous clusters and tumor diathesis are the distingushing features of stage Ib 2. It is nonethdess difficult to differentiate stage Ia from Ib 1.

View full abstract

We used an immunohistochemical technique to determine whether monoclonal antibody (PC 10) for proliferating cell nclear antigen (PCNA) could be useful as a marker of proliferating cells with formalin-fixed, paraffin-embedded tissue sections of 116 epithelial ovarian tumors. The mean labeling rate for PCNA was 33.0% in the malignant group (79 cases), 12.5% in the borderline-malignant group (17 cases) and 3.4% in the benign group (20 cases), showing significant differences among all groups. The mean labeling rate for PCNA in the malignant group was significantly higher, 35.0%, in the prechemotherapy group (61 cases) than in the postchemotherapy group (18 cases: 26.1%). These results suggest that the decrease in proliferating activity in the malignant cases with PCNA staining may be attributable to chemotherapy. Furthermore, there was a positive correlation (r=0.57) between the labeling rate for PCNA and mitotic activity. These results suggest that PCNA is a useful marker for distinguishing malignant tumors and assessing mitotic activity.

View full abstract

Sixty-six cervical specimens were obtained from thirtythree cases with squamous lesions of the cervix (ranging from moderate dysplasia to early invasive squamous carcinoma) with a Cervex-brush and cotton swab. Cytological diagnoses of the two groups were compared with final histological diagnoses. Correct cytodiagnosis was obtained in 25 Cervex-brush specimens (75.8%) and 12 cotton swab specimens (36.4%). Cytological underdiagnosis was recognized in 4 Cervex-brush specimens and 21 cotton swab specimens including three false negative cases in the cotton swab group. The quantity of endocervical cells (endocervical columnar cells plus squamous metaplastic cells) was significantly greater in Cervex-brush smears than that in cotton swab specimens. The rate of under cytodiagnosis was significantly higher in specimens lacking squamous metaplastic cells than in those with metaplastic cells. These results demonstrate that the cotton swab is not an adequate cytologic sampling technique for the detection of squamous lesions of the cervix.

View full abstract

Clinicocytopathological analyses of 18 cases with metastatic subcutaneous tumor which had been examined by aspiration cytology were done together with histological examination of resected materials and autopsy findings.
The 18 cases were classified into the two groups, Twelve patients had metastasis directly to subcutaneous tissue and the other 6 had subcutaneous invasion of metastatic bone tumor. Among the 18 cases, needle aspiration cytology was positive in 16 (88.9%), and suspicious in 2 (11.1%).
The 16 cases with correct cytological diagnosis consisted of 9 cases (56.3%) of adenocarcinoma, 2 (12.5%) hepatocellular carcinomas, and one case each of squamous cell carcinoma, large cell carcinoma, small cell carcinoma, transitional cell carcinoma, and Wilms' tumor. In 94% of cases, the histologic type of the tumor diagnosed by surgical or autopsy specimens was consistent with that of the cytological specimen.
Among the 16 cases with positive cytology and an identifiable primary site, cytologic examination made it possible to estimate histologic type and the primary site of the tumor in 6 cases (37.5%). These included well differentiated adenocarcinoma of the colon, small cell carcinoma of the lung, well differentiated papillary carcinoma of the thyroid, and the clear cell type of renal cell carcinoma. In the remaining 10 cases (62.5%), only the histologic type of the tumor could be estimated. Most were adenocarcinomas originating from the breast, lung, colon or stomach. Simultaneous examination of the histologic specimen taken from candidate lesions facilitated estimation of the primary site.

View full abstract

Fine needle aspiration cytology was performed on 19 patients with primary salivary gland tumors during the 1987-1991 period in our hospital. Diagnoses were histologically confirmed. Accuracy was 71.4% for benign tumors and 60% for malignant tumors. Cytologic diagnosis agreed with the histologic diagnosis in only 26.3% of cases.
A retrospective analysis of the cytologic smears was performed on the cases in which there was a disparity with histologic diagnosis. In the smears of benign pleomorphic adenoma, when either epithelial elements or stromal elements appeared alone in cytologic smears, these smears had a tendency to be diagnosed as benign lesions of the salivary glands. The main reason for the false positive diagnosis of benign pleomorphic adenoma was the appearrance of anisokaryosis and/or pleomorphism of tumor cells in the smears. But, as myxomatous stroma were found in the background, the diagnosis of benign pleomorphic adenoma had to be correct. In the smears of Warthin tumors, oncocytic cells were misdiagnosed as columnar cells. Cytologic recognition of a dirty background populated by lymphocytes, of a cellular arrangement resembling a honeycomb, and of greater cytoplasmic density and granularity was carefully sought. In two false negative cases of mucoepidermoid carcinoma, elements of squamous cells, intermediate cells and mucous-secreting cells were not consistently observed. Careful observation of slight hyperchromasia and deeply stained nucleoli was considered to be important. Seven out of 19 salivary gland tumor cases were diagnosed as swelling of cervical nodes by palpation. It was thus very difficult to distinguish tumor cells originating from the salivary glands.

View full abstract

We re-examined fine needle aspiration smears from 112 histologically proven cases of papillary carcinoma of the thyroid. Twenty cases (17.9%) revealed no malignant cells on the smears. Thirteen of these true negative cases were aspirated from normal thyroid tissue or adenomatous goiter near the papillary carcinoma. There were no aspirated cells on the smear in the remaining seven cases. Microcarcinoma (a tumor less than 1 cm in size), an association with calcification and/or ossification and totally or partially encapsulated types represent true negative cases. On the other hand, 16 cases (14.3%) were considered to have been under-diagnosed. The inaccurate diagnoses obtained in these cases may be attributable to inadequate sampling of the lesions or a paucity of intranuclear inclusions and nuclear grooves.

View full abstract

One thousand two hundred fifteen patients who visited at the breast clinic of Nagano Cancer Center on an out-patient basis, during the past 7 years and 3 months, underwent Aspiration Biopsy Cytology (ABC).
The true positive rate (class IV·V) for ABC among 295 breast cancers in these patients was 86.1% and the true negative rate (class I·II) for 917 benign breast diseases was 83.3%.
The Proportion of Class III determinations in each groups resulted in 4.7%(14 cases) and 6.8%(62 cases), respectively.
A total of 76 cases which had been diagnosed as class III by ABC, was divided into two groups, namely, a class III a group (51 cases) and a class III b group (25 cases).
The following results were obtained by comparing these two groups.
1) The frequency of class III b in the class III groups was the highest (64.3%) for breast cancer, followed by mastopathy (40.9%), while the frequency for fibroadenoma was the lowest (9.5%).
2) The frequency of breast cancer in the class III a groups was 9.8% and was 36.5% in the class III b groups.
3) Among class III a and class III b groups which had been diagnosed as breast cancer by palpation, the frequency of breast cancer was not significantly different, but in the groups diagnosed as mastopathy, this frequency was higher in class III b than in class III a groups.
4) In the groups which had been diagnosed as behigh (I·II) by mammography, among class III a cases there were no breast cancers, but, approximately 1/3 of class III b cases did have breast cancer. In the groups which had been diagnosed as malignant (IV·V) by mammography, there were higher possibilities of breast cancer than even in class III a.
5) In the groups which had been diagnosed as benign to suspicious (I·III) by echography, among class III a cases there were few breast cancers, but, approximately 30% of class III b cases did have breast cancer. In the groups which had been diagnosed as malignant (IV·V) by echography, all cases had breast cancer independent of whether they were class III a or class III b.
6) In the groups which had been diagnosed as benign (I·II) by thermography, among class III a cases there were few breast cancers, but approximately 1/3 of class III b case did have breast cancer. In the groups which had been diagnosed as malignant (IV·V) by thermography, half of the class III b cases had breast cancer.
The conclusion can be drown from the above results, that those cases diagnosed as class III by ABC were divisible into class III a and class III b groups.

View full abstract

Nuclear DNA ploidy levels were examined in 47 cases of mucus-producing adenocarcinoma of the lung resected during the last 19 years, and were compared in relation to the prognoses of these patients. Twenty-five cases of goblet cell type adenocarcinoma, were divided into 16 cases (64%) with DNA diploidy and 9 cases (36%) with DNA aneuploidy patterns. Twenty-two cases of bronchial gland cell type adenocarcinoma were classified into 3 cases (14%) of DNA diploidy and 19 cases (86%) of DNA aneuploidy. In these two groups, there were remarkable differences in the distribution of DNA ploidy level even in the same group of mucusproducing tumors of the lung. In goblet cell type adenocarcinoma, the 5-year survival rates in groups with DNA diploidy and DNA aneuploidy were 71.4% and 62. 5%, respectively. In bronchial gland cell type adenocarcinoma, the 5-year survival rates in groups with DNA diploidy and DNA aneuploidy were 33.4% and 48.3%, respectively. These results indicate that there is no significant difference between DNA ploidy levels in terms of the prognosis of mucusproducing adenocarcinoma. Nevertheless, in the diffuse type of goblet cell type adenocarcinoma, cases with DNA aneuploidy showed a poorer prognosis than did those with DNA diploidy.

View full abstract

The authors studied the imprint cytology in two cases of small cell carcinoma (SCC) of the gall bladder. In one case, and intraopeative speciner was used for imprinting, and in the other case, a postoperative specimen was used. Although both cases had the classical cell clusters of SCC, with nuclear moldings and indian filings, the overall cytological features were distinctly different. The intraoperative case had numerous cell clusters, many of them resembling those observed in poorly differentiated adenocarcinoma, whereas the postoperative case contained innumerable small, round, isolated, cells, resulting in some resemblance to atypical carcinoid or malignant lymphoma in many areas. Histologically, the intraoperative case was a SCC with squamous metaplasia and good tissue presaervation, and the postoperative case was a combined SCC with adenocarcinoma localized in overlying areas of SCC, and had an autolytic nature. Both cases were positive for Grimelius stain and for neuron-specific-enolase (NSE) immunohistochemical stain in SCC areas. Based on these findings, the authors consider that the difference in cytological features between the two is mainly attributable to the quality of tissue preservation. Autolysis might prevent accurate interpretation of imprint cytology in SCC. Cytotechnologists and cytopathologists should realize that larger surgical specimens may have varying degrees of autolysis before their arrival at the pathology laboratory. To our knowledge, this is the first cytological report of SCC of the gall bladder in the literature.

View full abstract

The case of a 68-year-old female, who had suffered from liver cirrhosis for several years before the discovery of hepatocellular carcinoma (HCC) at the begining of 1991, is reported. Transcatheter arterial embolization (TAE) was performed in March, and again in November, of 1991, Because a new nodule of HCC was discovered, she was admitted for a third course of treatment in March, 1992. After a admission the patient's condition took a sudden turn for the worse and ascites was noticed.
Cytologic examination of the ascites revealed numerous malignant cells with increased nuclear-cytoplasmic ratios, peripherally located nuclei, variations in nuclear size and shape, thickened nuclear membranes, granular chromatin patterns, prominent nucleoli and vacuolated cytoplasm. Multinucleated giant malignant cells were also observed. The cytologic diagnosis of ascites was poorly differentiated HCC. Subsequent autopsy revealed peritoneal dissemination of poorly differentiated HCC.
Poorly differentiated adenocarcinoma, amelanotic melanoma, malignant lymphoma and other malignant cells should be excluded in making a cytologic diagnosis of ascites with HCC.

View full abstract

Cytological characteristics in fine needle aspiration cytology of secretory carcinoma of the breast in a 29-year-old woman are reported. Fine needle aspirates disclosed tumor cells possessing a predominantly rich, pale lacy cytoplasm and round to oval nuclei without marked atypia. Vacuoles or mucoid materials of various sizes were often encountered in the cytoplasm with occasional formations of intracytoplasmic lumina (ICL). The mucoid substance was positive on periodic acid-Schiff, alcian blue and mucicarmine stains. In addition, characteristic structures consisting of central mucoid material surrounded by two or more tumor cells and the larger aggregated forms of such units were found. These structures, especially the aggregated forms, were very few in number in the touch smear specimen. Furthermore, the electron microscopic study failed to disclose any peculiar structures, except for the intercellular lumina. Therefore, these structures in fine needle aspiration of a secretory carcinoma may represent three-dimensional unit constituents of the tumor. We conclude that cytological diagnosis of secretory carcinoma of the breast is possible on the basis of these characteristic structures and positive staining for epithelial mucin.

View full abstract

A case of true malignant mixed tumor of the right parotid gland is reported. The parotid tumor had started to grow rapidly after an approximately 30-year quiescent period. Surgical resection was performed in September 1988. The resected tumor, measuring 5×3×3 cm, contained both carcinomatous and sarcomatous elements, the former consisting of tubular adenocarcinoma, the latter of chondrosarcoma with osteosarcoma. A myxoid matrix, a remnant of pleomorphic adenoma, could be also identified. Scratch cytology of the tumor revealed that both elements were clusters of atypical epithelial cells and atypical round cells in the lacunae within the avascular and myxoid matrix.
It is assumed from these clinical, histological, and cytological findings that the tumor was derived from a pre-existing pleomorphic adenoma. As to the histogenesis of the true malignant mixed tumor, it has been thought that both carcinomatous and sarcomatous elements might originate from myoepithelial cells. In the present case, plasmacytoid myoepithelial cells containing many cytoplasmic filaments were ultrastructurally identified within the sarcomatous area.

View full abstract

An autopsy case of meningeal carcinomatosis originating from lung cancer is reported. The patient, a 61-year-old male, complained of numbness in the right upper limb, speaking disturbance, headache and vertigo, Meningeal carcinomatosis was diagnosed by cytological examination of the cerebrospinal fluid (CSF).
The cytology revealed adenocarcinoma cells with pseudo-cilia, which are generally considered to be characteristic of ovarian cystadenocarcinoma. Though the primary lesion was investigated, it could not be found before death. Autopsy revealed minute pulmonary tumors, histologically determined to be papillary adenocarcinoma. Pseudo-cilia in the CSF cytology of a primary lung adenocarcinoma have not previously been described. Upon recognizing malignant cells with pseudo-cilia, lung cancer should be considered in the differential diagnosis.

View full abstract