Nuclei Of Malignant Cells Research Articles

Chromatin condensed domains revealed by AFM, and their transformation in mechanically deformed normal and malignant cell nuclei

It has been generally accepted that heterochromatin is represented by a regular, dense and closed structure, while euchromatin is open and sparse. Recent evidence indicates that chromatin is comprised of irregular nucleosome clutches compacted within the nucleus. Transcriptional events transform the chromatin architecture, resulting in appearance of 100–300 nm nucleosomal aggregates. Meanwhile, the current paradigm of chromatin architecture is largely fragmented. In this communication, we unraveled chromatin ultrastructure of normal and malignant cell nuclei through mechanical deformation of the nuclei and Atomic Force Microscopy (AFM) analysis of the resulting landscape. In human skin fibroblasts cell nuclei, nanodomains of about 16.5–33.5 nm were revealed. Hierarchical folding of the chromatin of normal nuclei was observed: the nanodomains formed irregular fiber-like structures that coalesced into the macroscale chromatin compartments. In fibrosarcoma cell nuclei DNA supercoiling domains (SDs) of about 66.3–113.0 nm, uniformly distributed within the nuclei, were revealed. Transformation of the morphology of the condensed chromatin domains through up- and downregulation of supercoiling was demonstrated.

Aptamer/Peptide-Functionalized Genome-Editing System for Effective Immune Restoration through Reversal of PD-L1-Mediated Cancer Immunosuppression.

Effective reversal of tumor immunosuppression is of critical importance in cancer therapy. A multifunctional delivery vector that can effectively deliver CRISPR-Cas9 plasmid for β-catenin knockout to reverse tumor immunosuppression is constructed. The multi-functionalized delivery vector is decorated with aptamer-conjugated hyaluronic acid and peptide-conjugated hyaluronic acid to combine the tumor cell/nuclear targeting function of AS1411 with the cell penetrating/nuclear translocation function of TAT-NLS. Due to the significantly enhanced plasmid enrichment in malignant cell nuclei, the genome editing system can induce effective β-catenin knockout and suppress Wnt/β-catenin pathway, resulting in notably downregulated proteins involved in tumor progression and immunosuppression. Programmed death-ligand 1 (PD-L1) downregulation in edited tumor cells not only releases the PD-1/PD-L1 brake to improve the cancer killing capability of CD8+ T cells, but also enhances antitumor immune responses of immune cells. This provides a facile strategy to reverse tumor immunosuppression and to restore immunosurveillance and activate anti-tumor immunity.

Comparison of Fixation Methods for Preservation Cytology Specimens of Cell Block Preparation Using 10% Neutral Buffer Formalin and 96% Alcohol Fixation in E-cadherin and Ki-67 Immunohistochemical Examination.

BACKGROUND:Cytological and molecular examinations are among the most important examinations in cancer diagnosis. 96% alcohol is a fixative solution commonly used by clinicians for cytological samples because of its accessibility and affordability. Cellblock preparation from cytology specimen may increase morphology detail and may be used for further biomarker analysis. E-cadherin is an adhesion protein expressed in the cell membrane of most carcinoma. Ki67 is a protein expressed in nuclei of malignant cells that used as a proliferation marker.AIM:This study was designed to investigate the effect of fixation duration in 96% alcohol on protein preservation for immunohistochemistry (IHC) evaluation compared to 10% neutral buffered formalin (NBF) as the gold standard.METHODS:Twenty-five fine-needle aspiration biopsy (FNAB) specimen diagnosed as carcinoma were fixed in 10% NBF and 96% alcohol for 1 hour, 6 hours, 24 hours, 48 hours and 72 hours. Cell blocks preparation were made from those 6 groups of specimens. E-cadherin and Ki67 IHC were done to cell blocks section and evaluated. The data were statistically analysed using the Friedman test with p-value < 0.05 of a significant level.RESULTS:There were significant differences between E-cadherin and Ki67 expression in cell block preparation from 96% alcohol-fixed cytology specimen for 1 hour, 6 hours, 24 hours, 48 hours and 72 hours to 10% NBF (p = 0.0001).CONCLUSION:The result indicated that 96% alcohol is not suitable as a fixative solution for cell block preparation in E-cadherin and Ki-67 IHC examination.

Abstract 2451: The nuclear-targeted gold nanoparticles inhibit the tumor growth in mouse model as potential anticancer agents

Abstract The use of gold nanoparticles (AuNPs) as chemotherapeutic agents, based on their intrinsic antineoplastic properties is becoming an increasingly exciting possibility for the treatment of cancer. The specific targeting of AuNPs to the nucleus of malignant cells can be achieved via peptide functionalization of the nanoparticle surface, resulting in DNA damage and subsequent apoptosis. Here, we utilized these novel AuNPs for systemic delivery in vivo and determined their efficacy as chemotherapeutic agents. In addition, we assessed their effects on overall health in order to characterize their systemic toxicity. AuNPs (30 nm in diameter) were functionalized with an Arg-Gly-Asp (RGD) peptide sequence, providing tumor cell specific targeting, as well as a Lys-Lys-Lys-Arg-Lys nuclear localization sequence (NLS) peptide. These novel nuclear-targeting AuNPs were delivered via tail vein injection in nude mice bearing Tu212 tumors. Their in vivo therapeutic efficacy was evaluated, along with assessing their systemic toxicity. The AuNPs, specifically targeted to the tumor cell nucleus, exhibit circulation half-life of 4.1 h, which is exceptionally greater than common chemotherapeutic agents and, although they exhibit uptake by organs of the reticuloendothelial system (RES) (e.g. liver and spleen), they also accumulate in the tumor. In addition, when systemically delivered at low doses (5 nM AuNPs) these novel targeted AuNPs show significant tumor regression (via DNA damage and subsequent apoptosis) when compared to their non-targeting counterparts (P = 0.004), with no observed toxicity in terms of hepatic function, renal function, or body weight. In conclusion, by delivering AuNPs to the nucleus of tumor cells, at low doses in vivo, high therapeutic efficacy is achieved with minimal side effects, indicating the potential for the use of these novel nuclear-targeted AuNPs as chemotherapeutic agents. Citation Format: Xianghong Peng, Megan A. Macke, Hyung Ju C Shin, Sreenivas Nannapaneni, Nelson Chen, Sungjin Kim, Zhuo (Georgia) Chen, Mostafa A El-Sayed, Dong Shin. The nuclear-targeted gold nanoparticles inhibit the tumor growth in mouse model as potential anticancer agents. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2451. doi:10.1158/1538-7445.AM2015-2451

A Cell-permeable Stat3 SH2 Domain Mimetic Inhibits Stat3 Activation and Induces Antitumor Cell Effects in Vitro

Given the role of constitutively active Signal Transducer and Activator of Transcription (Stat) 3 in human tumors, Stat3 inhibitors would be useful as novel therapeutics and as tools for probing Stat3-mediated tumor processes. We herein report that a 28-mer peptide, SPI, derived from the Stat3 SH2 domain, replicates Stat3 biochemical properties. Studies show SPI and Stat3 (or Stat3 SH2 domain) bind with similar affinities to known Stat3-binding phosphotyrosine (pY) peptide motifs, including those of the epidermal growth factor receptor (EGFR) and the high-affinity, IL-6R/gp130-derived pY-peptide, GpYLPQTV-NH(2). Consequently, SPI functions as a potent and selective inhibitor of Stat3 SH2 domain:pTyr interactions and disrupts the binding of Stat3 to the IL-6R/gp130 peptide, GpYLPQTV-NH(2). Fluorescence imaging and immunofluorescence staining/laser-scanning confocal microscopy show SPI is cell membrane-permeable, associates with the cytoplasmic tail of EGFR in NIH3T3/hEGFR, and is present in the cytoplasm, but strongly localized at the plasma membrane and in the nucleus in malignant cells harboring persistently active Stat3. Moreover, SPI specifically blocks constitutive Stat3 phosphorylation, DNA binding activity, and transcriptional function in malignant cells, with little or no effect on the induction of Stat1, Stat5, and Erk1/2(MAPK) pathways, or on general pTyr profile at the concentrations that inhibit Stat3 activity. Significantly, treatment with SPI of human breast, pancreatic, prostate, and non-small cell lung cancer cells harboring constitutively active Stat3 induced extensive morphology changes, associated with viability loss and apoptosis. Our study identifies SPI as a novel molecular probe for interrogating Stat3 signaling and that functions as a selective inhibitor of Stat3 activation with antitumor cell effects.

Expression of androgen receptor in bladder urothelial carcinoma and its clinical significance

Objective To investigate the expression of androgen receptor (AR) in bladder urothelial carcinoma and to explore its clinical significance. Methods Immunohistochemical SP method was employed to detected the expression of AR in bladder cancer tissue (123 cases) and the normal controls (18 cases) . In addition,clinicopathological data of them were analyzed. Results AR protein was mainly localized at cellular nucleus of malignant cells. The positive expression rate of AR in bladder cancer tissue was 46.3% ,but low positive expression rate (5.6% ) was found in normal bladder tissue. The expression of AR was markedly associated with tumor pathological grade and clinical stage ( P < 0.05 ) , but not with age and sex. Moreover,the expression of AR was observed more frequently in cases with lymph node metastasis. Conclusion The expression of AR is associated with differentiation and invasion in bladder urothelial carcinoma, it implies that AR might play important roles in the development and progression of bladder cancer. Key words: Bladder neoplasms; Androgen receptor; Immunohistochemical

Immunohistochemical Expression of Estrogen and Progesterone Receptors Identifies a Subset of NSCLCs and Correlates with EGFR Mutation

To determine the frequency of estrogen receptor alpha and beta and progesterone receptor protein immunohistochemical expression in a large set of non-small cell lung carcinoma (NSCLC) specimens and to compare our results with those for some of the same antibodies that have provided inconsistent results in previously published reports. Using multiple antibodies, we investigated the immunohistochemical expression of estrogen receptors alpha and beta and progesterone receptor in 317 NSCLCs placed in tissue microarrays and correlated their expression with patients' clinicopathologic characteristics and in adenocarcinomas with EGFR mutation status. Estrogen receptors alpha and beta were detected in the nucleus and cytoplasm of NSCLC cells; however, the frequency of expression (nucleus, 5-36% for alpha and 42-56% for beta; cytoplasm: <1-42% for alpha and 20-98% for beta) varied among the different antibodies tested. Progesterone receptor was expressed in the nuclei of malignant cells in 63% of the tumors. Estrogen receptor alpha nuclear expression significantly correlated with adenocarcinoma histology, female gender, and history of never smoking (P = 0.0048 to <0.0001). In NSCLC, higher cytoplasmic estrogen receptor alpha expression significantly correlated with worse recurrence-free survival (hazard ratio, 1.77; 95% confidence interval, 1.12, 2.82; P = 0.015) in multivariate analysis. In adenocarcinomas, estrogen receptor alpha expression correlated with EGFR mutation (P = 0.0029 to <0.0001). Estrogen receptor beta and progesterone receptor but not estrogen receptor alpha expressed in the normal epithelium adjacent to lung adenocarcinomas. Estrogen receptor alpha and beta expression distinguishes a subset of NSCLC that has defined clinicopathologic and genetic features. In lung adenocarcinoma, estrogen receptor alpha expression correlates with EGFR mutations.

Prognostic value of microsatellite instability determined by immunohistochemical staining of hMSH2 and hMSH6 in urothelial carcinoma of the bladder

Mismatch repair (MMR) genes are involved in the recognition and repair of acquired DNA damage, which arises during cell division, thus playing an essential role in preserving genetic stability. Immunohistochemistry was applied to 130 specimens from urothelial carcinoma (UC) of the bladder to detect expression of MMR gene products hMSH2 and hMSH6, and to investigate its clinicopathological and prognostic value. hMSH2 and hMSH6 protein expression was exclusively detected in the nuclei of malignant cells. Of the 112 cases evaluable for hMSH2, 29 (25.9%) were negative and of the 130 UCs evaluable for hMSH6, 64 (49.2%) were negative, and were thus considered to depict MSI. Nuclear hMSH2 values were statistically lower in non-invasive UCs (Ta-T1) (p=0.013) and in carcinomas with decreased p53 staining (p=0.04). Lower hMSH6 values were more often met in well-differentiated tumors (p<0.0001) and in tumors with low expression of p53 (p=0.016), topoIIalpha and caspase 3 (p=0.017 and p=0.018, respectively). Both hMSH2- and hMSH6-negative immunoreactions were found to have a favorable impact on overall patient survival (p=0.041 and p=0.034, respectively), this finding being further verified in the multivariate analysis of hMSH2 (p=0.026). This is the first study to show that lack (and not reduction designated according to various cut-off points) of hMSH2 and hMSH6 correlated with non-invasive tumors of lower grade and is of favorable prognostic significance in patients suffering from bladder carcinoma.

Endoscopic ultrasound-guided fine-needle aspiration cytology diagnosis of solid pseudopapillary tumor of the pancreas: A case report and literature review

We describe the clinical, imaging and cytopathological features of solid pseudopapillary tumor of the pancreas (SPTP) diagnosed by endoscopic ultrasound-guided (EUS-guided) fine-needle aspiration (FNA). A 17-year-old woman was admitted to our hospital with complaints of an unexplained episodic abdominal pain for 2 mo and a short history of hypertension in the endocrinology clinic. Clinical laboratory examinations revealed polycystic ovary syndrome, splenomegaly and low serum amylase and carcinoembryonic antigen (CEA) levels. Computed tomography (CT) analysis revealed a mass of the pancreatic tail with solid and cystic consistency. EUS confirmed the mass, both in body and tail of the pancreas, with distinct borders, which caused dilation of the peripheral part of the pancreatic duct (major diameter 3.7 mm). The patient underwent EUS-FNA. EUS-FNA cytology specimens consisted of single cells and aggregates of uniform malignant cells, forming microadenoid structures, branching, papillary clusters with delicate fibrovascular cores and nuclear overlapping. Naked capillaries were also seen. The nuclei of malignant cells were round or oval, eccentric with fine granular chromatin, small nucleoli and nuclear grooves in some of them. The malignant cells were periodic acid Schiff (PAS)-Alcian blue (+) and immunocytochemically they were vimentin (+), CA 19.9 (+), synaptophysin (+), chromogranin (-), neuro-specific enolase (-), a1-antitrypsin and a1-antichymotrypsin focal positive. Cytologic findings were strongly suggestive of SPTP. Biopsy confirmed the above cytologic diagnosis. EUS-guided FNA diagnosis of SPTP is accurate. EUS findings, cytomorphologic features and immunostains of cell block help distinguish SPTP from pancreatic endocrine tumors, acinar cell carcinoma and papillary mucinous carcinoma.

Cytological features of cervical smears in serous adenocarcinoma of the endometrium.

Serous adenocarcinoma (SAC) of the endometrium has a poor prognosis compared with that of typical endometrioid adenocarcinoma (EAC). The objective of this study was to determine whether SAC can be distinguished from EAC preoperatively by cervical or endometrial cytology. Cervical smears and endometrial smears obtained from 128 patients with endometrial carcinoma were reviewed. Histological types included 117 cases of EAC and 11 cases of SAC. The positive rates of cervical smears and those of endometrial smears in SAC and EAC cases were compared. Papillary clusters and bare nuclei of malignant cells in positive cervical smears were also investigated for their diagnostic significance in discriminating between EAC and SAC. The positive rate of cervical smears in SAC was significantly higher than that in EAC (72.7 vs 27.4%, P < 0.05). Among cases with positive cervical smears, there were significantly more cases with papillary clusters and/or bare nuclei in cases of SAC than in cases of EAC. When endometrial carcinoma is clinically suspected and a cervical smear is positive, the predominance of either papillary clusters or features of bare nuclei of malignant cells in the smear may indicate the presence of SAC.

New texture features based on the complexity curve

A new set of texture features based on the complexity curve is proposed for texture analysis. The complexity curve is obtained by computing the number of black-to-white transitions observed in a binary image for all possible threshold values in a gray-level texture image. The new features characterize the first-order statistics of the complexity curve, and therefore give a second-order description of texture. The set of features is applied to texture classification, and to capture the directionality and the periodicity of texture. For texture classification, two experiments are performed. First, some Brodatz textures are used to evaluate the classification performance of the features. Some of the traditional texture features, extracted from the co-occurrence matrix, have also been used for comparison. Secondly, the features are applied to the classification of Transmission Electron Micrographs of premalignant and malignant cell nuclei in a controlled mouse liver carcinogenesis experiment. Directionality and periodicity are two high-level texture features that guide the process of perceptual grouping of texture images. Therefore, two additional experiments have been performed to evaluate the performance of the complexity curve features in estimating the texture directionality and periodicity. The experimental results demonstrate that the new texture features based on the complexity curve are very useful tools for texture analysis.

C-Src protein expression is increased in human breast cancer. An immunohistochemical and biochemical analysis.

In human breast cancer, c-Src activity is elevated compared to normal breast tissue. It is not yet known whether this increase in c-Src activity is accompanied by an increase in c-Src protein expression. In this study, c-Src activity and protein expression were determined in a series of human breast cancers and in normal breast tissue, using immune complex kinase assays and immunoblotting. As the heterogeneity of breast cancer is not taken into account in these biochemical experiments, immunohistochemistry was also used to distinguish between normal and malignant cells. In human breast cancers, the c-Src activity is increased 4- to 30-fold, compared with normal breast tissue. This enhanced activity is accompanied by an increase in c-Src protein expression, as shown by both immunoblotting and immunohistochemistry. Immunohistochemistry indicates that the majority of c-Src appears to be concentrated around the nucleus in malignant cells, whereas in normal cells, it is distributed more evenly in the cytoplasm. These data confirm that c-Src activity is increased in human breast cancer. In addition, this study provides strong immunohistochemical evidence that the c-Src protein is also overexpressed, enabling a distinction to be made between normal and malignant cells.

Interphase ribosomal RNA cistron staining in thyroid epithelial cells in Grave's disease, Hashimoto's thyroiditis and benign and malignant tumours of the thyroid gland

Aim-To evaluate the expression of ribosomal cistrons in human thyroid epithelial cells (TECs) of patients with Grave's disease, Hashimoto's thyroiditis and benign and malignant tumours of the thyroid gland.Methods-TEC nucleoli were investigated in fine needle biopsy specimens from 10 controls, 39 patients with Grave's disease, 15 with Hashimoto's thyroiditis, 56 with benign, and 15 with malignant tumours of the thyroid. A one step silver staining method was applied. In most cases serum concentrations of thyroxine and triiodothyronine as well as goitre size were determined. In every case 100 TECs were evaluated for the mean numbers of nucleoli and for the average number of argyrophilic nucleolar organiser regions (AgNORs) per nucleus.Results-NORs were activated in all patients, but not in controls. The numbers of AgNORs in patients with Grave's disease were closely correlated with thyroxine or triiodothyronine, or both, concentrations and with the size of the thyroid. In patients with Hashimoto's thyroiditis about 30% of TECs nucleoli did not contain AgNORs, whereas others were heavily impregnated with silver. Compared with controls and benign tumours, the nucleoli of carcinomatous TECs were larger and irregular in shape. The mean number of AgNORs per nucleus in malignant cells was higher than that in their benign counterparts.Conclusions-The mechanism by which NORs are activated in TECs varies depending on the type of lesion. The higher AgNOR score in TECs from malignant tumours can be used to distinguish them from their benign counterparts.

Karyometry of malignant melanoma cells present in skin strippings.

Skin strippings harvest the malignant melanoma cells eliminated through the stratum corneum. In this study, morphometry was applied to such non-invasive samplings in order to derive objective information about nuclear atypia. Cyanoacrylate skin surface strippings were collected from 140 malignant melanomas. Karyometry was performed to quantify the size and shape of malignant cell nuclei. Anisokaryosis was prominent although the outline of nuclei were rather smooth. Image analysis of skin strippings harvested from malignant melanomas shows that anisokaryosis is the major cytologic feature of malignant melanoma cells in transit in the stratum corneum.

Cell proliferation and apoptosis: Immunohistochemical evidence of p53 protein in human placenta and choriocarcinoma cell lines

We investigated the expression of the tumour-suppressor and cell cycle control protein p53 in human first trimester and term placenta, three choriocarcinoma cell lines (Jeg-3, JAR, BeWo) and human choriocarcinoma. Using monoclonal antibodies against p53 (DO-7, Ab-6, DO-1, PAb 1801), paraffin-embedded sections of first trimester and full-term placentae, human choriocarcinoma and Jeg-3, JAR and BeWo, as well as cytospins, were evaluated immunohistochemically. In addition, Western blots were carried out with the same antibodies on choriocarcinoma cell lines. In placentae, a small number of villous and extravillous cytotrophoblast cells, as well as very few syncytiotrophoblast cells, stained intensively. Also, p53 was visualized in some nuclei of the placental basal plate, whereas stroma and endothelium were negative for p53. Jeg-3, JAR and BeWo also showed a positive nuclear reaction with all applied antibodies. In paraffin-embedded sections of human choriocarcinoma, staining was confined to the nuclei of malignant cells. The results suggest that p53 is overexpressed not only in malignant tumour cells but in certain trophoblast cell populations of the human placenta as well.

AN “ASYMPTOTIC FRACTAL” APPROACH TO THE MORPHOLOGY OF MALIGNANT CELL NUCLEI

To investigate quantitatively nuclear membrane irregularity, 672 nuclei from 10 cases of oral cancer (squamous cell carcinoma) and normal cells from oral mucosa were studied in transmission electron micrographs. The nuclei were photographed at ×1400 magnification and transferred to computer memory (1 pixel=35 nm). The perimeter of the profiles was analysed using the “yardstick method” of fractal dimension estimation, and the log-log plot of ruler size vs. boundary length demonstrated that there exists a significant effect of resolution on length measurement. However, this effect seems to disappear at higher resolutions. As this observation is compatible with the concept of asymptotic fractal, we estimated the parameters c, L and Bm from the asymptotic fractal formula Br=Bm {1+(r/L)c}−1, where Br is the boundary length measured with a ruler of size r, Bm is the maximum boundary for r→0, L is a constant, and c=asymptotic fractal dimension minus topological dimension (D−Dt) for r→∞. Analyses of variance showed c to be significantly higher in the normal than malignant cases (P&lt;0.001), but log(L) and Bm to be significantly higher in the malignant cases (P&lt;0.001). A multivariate linear discrimination analysis on c, log(L) and Bm re-classified 76.6% of the cells correctly (84.8% of the normal and 67.5% of the tumor). Furthermore, this shows that asymptotic fractal analysis applied to nuclear profiles has great potential for shape quantification in diagnosis of oral cancer.

On the topology of normal chromatids and on their translocations in myelogenous leukemia

After some comments on the topology of chromatids, restructuring of the interphase nucleus is conjectured to depend upon the nuclear vesicle apparatus. These vesicles change the intrinsic shape of chromatids to fit the different topology of the interphase nuclear spheroid. Reciprocal translocations between selected chromatids result whenever the nucleus of malignant cells organizes de novo certain exceptional or emergency differentiation paths. However, the almost unavoidable chimeric genes resulting from these translocations may be less ominous than hitherto suspected. This seems to be the case for chronic myelogenous leukemia, where the bcr-abl chimeric gene lessens the aggressiveness of the primary clone when functioning in the context of myelomonocitic differentiation. Finally, our model estimates the statistical incidences of the bcr-abl chimera. These estimates are found to agree with clinical data better than evaluations from the random mutation theory.

P53 expression in breast cancer

Immunohistochemical evaluation of 200 primary breast cancers with the anti-p53 mouse monoclonal antibody (MAb) PAb421 showed positivity in nuclei of malignant cells in 31 cases (15.5%). PAb421+ cases were significantly more frequently epidermal growth factor receptor (EGF-R)-positive (67.7%; p less than 0.001) and estrogen receptor (ER)-negative (73.3%; p less than 0.001); they displayed surface histocompatibility class-1 (80.6%; p less than 0.01) and 11 (74.2%; p less than 0.05) antigens. Low values for progesterone receptor (mean 67.20 +/- 25.2 fmol/mg; p less than 0.05) and a high number of cells positive for the proliferation-associated antigen Ki-67 (log mean 6.88 +/- 0.33; p less than 0.01) were found in PAb421+ tumors as well as a high number of grade-3 infiltrating duct carcinomas (70%; p = 0.01). Of the 200 cases of mammary carcinoma, 88 were further analyzed using another human specific anti-p53 MAb PAb1801, and 40 (45.5%) were found positive. This MAb stained all the PAb421+ cases and was significantly associated with negative ER status (39.5%; p less than 0.05) and high Ki-67 scores (log mean 6.93 +/- 0.24; p = 0.001). Analysis of PAb1801+/Pab421- cases for HLA antigens, EGF-R and ER showed a phenotype similar to that of the p53-ve/ER+ carcinomas, except for the high Ki-67 score. No differences in age of the patient, number of involved nodes, tumor size, ploidy or labelling index scores were evident between p53+ and carcinomas. We concluded that p53 in mammary carcinomas is associated with ER-negative, growth factor receptor-positive, high-grade tumors, and is a promising new parameter to evaluate the cellular biology and prognosis of breast cancer.

Cytologic changes of cervical cancer according to the degree of invasion.

Cellular samples (cervical scrapings) obtained with wooden spatulas from patients with carcinoma in situ, early invasive cancer, and advanced cancer of the uterine cervix were examined. The number of malignant cells in the preparation increased with extent of the lesion. The granular chromatin structures of the nuclei of abnormal cells were observed in almost all cases, and the occurrence of irregularly distributed coarsely granular chromatin increased gradually with the extent of the cancer. Hyperchromasia of malignant cell nuclei and irregular thickening of the nuclear envelope were seen in almost all cases of the lesions. The occurrence of malignant and dysplastic cells of the deep layer types decreased with progression of the lesion from carcinoma in situ to early invasive cancer, and to advanced cancer. There was also a relative increase in the ratio of malignant cells to dysplastic cells.

子宮膣部重層扁平上皮のPapanicolaou染色における顕微分光的研究

Since the report of G. Papanicolaou, cytodiagnostics have been utilized for earier discovery of cancers in various medical fields. Particularly, in obstetrics, with the popularity of public health check by cytodiagnostics for earlier discovery of the cancer of uterine cervix, attention has been paid to the deep stainability of nucleus since it is most indicative of anomaly among all malignant findings related to cancer.The authors, with the purpose of obtain an objective standard for judgement of cytodiagnosis, had studied the method of quantitative determination of stainability (transmission)of hematoxilin-positive substance in the cell nucleus, by utilizing microspectrophotometric method reported by Caspersson et al in 1936.Scraping smear specimen was prepared by using th e cells from the cases aged 23 to 62 having normal cervics, inflammation, dysplasia, ca. in situ and invasive carcinoma which were discovered by histological diagnosis. The specimen was stained by Papanicolaou's method and the transmission of normal and abnormal squamous ephitheria (4785pieces) were comparatively studied by using Zeiss visible microspectrophotometer by visible light of a fixed wave length of 563 mμ since the separation width of cytoplasm and nucleus was largest and stable at this wave length. The study revealed that the transmission of various kinds of nucleus of normal cell was 0.27-0.85 for superficial cells (2187 pieces),0.28-0.83 for intermediate cells (492 pieces) and 0.170.79 for parabasal cells (1045 pieces). While it was 0.15-0.62 for dyslaryotic cells (495 pieces)O.03-0.75 for malignant cells (1066 pieces).The study on the rate of appearance of various cell nuclei revealed that while sufficient separation was noticed between malignant cell nuclei and the nuclei of superficial, intermediate, parabasal and dyskaryotic cells, the separation between dyskaryotic nuclei and the nuclei of superficial, intermediate nuclei and the nuclei of superficial, intermediate and parabasal cells was not clear. When the rate of appearance was compared between various classes of cells according to Papanicolaou's classification, no cell nuclei indicating transmission of less than 0.20 was recognized for benign group (PC I, II) and no nuclei of less than 0.10 was noticed for boundary was noticed for boundary group (PC III). Whereas with malignant group (PC IV, V), the rate of appearanc of cell nuclei indicating less than 0.10 of transmission was 5.3-22.6 %. Based on these experimental data, the standard was established by which the unknown specimen indicating the minimum value (t) of transmission of more than 0.21 (0.20<t) was regarded benign group, those indicating 0.11 to 0.20 (0.10<t<0.20) were regarded boundary group and those indicating less than 0.10 (0.10>t) were regarded malignant group. Based on such standard, blind test was conducted on the sample of 24 cases. The result was righteous in 22 cases except one false negative and one false positive cases. It was thus confirmed that the transmission of the light of a fixed wave length through the cells is sufficiently useful as an important objective and quantitative standard for judgement, when it is co-used with nucleus diameter and N/C ratio.