Detection of chemosensitivity using K18-Asp(396) (M30) antibody in HeLa and OVCAR-3 cell lines treated with anticancer agents / 대한산부인과학회지

OBJECTIVE: The aim of this study was to detect the levels of M30-antigens as a biomarker of apoptosis in cells and their culture media after treatments with anticancer drugs as a preclinical study. METHODS: After HeLa and OVCAR-3 cells were treated respectively with paclitaxel, cisplatin, and camptothecin, the harvested cells were stained sequentially with M30 monoclonal antibodies and propidium iodide (PI). Afterwards, they were analyzed using a FACScan flow cytometer and observed under an immunofluorescence microscope for M30-FITC immunofluorescences. Levels of M30 antigens were also detected in their culture media using M30-Apoptosense ELISA kit. RESULTS: The levels of M30-FITC immunofluorescences were elevated in both cell lines after each drug treatments compared with those of control cells. The levels of M30 antigens detected by ELISA in media culturing each cell line treated with each of drugs were elevated compared with those of control cells. CONCLUSION: This study suggests that M30-antigens representing chemotherapy induced apoptosis may be a useful biomarker for predicting and monitoring the response of neoadjuvant chemotherapy in patients with gynecologic cancers.

Measurement of apoptosis using M30 in culture media of cell lines treated with anti-cancer agents / 대한산부인과학회지

OBJECTIVE: We investigated a possible use of the induced apoptosis as a biomarker in the cells and their media treated with commonly used anti-cancer agents in gynecologic malignancies. METHODS: After treatments with low and high concentrations of paclitaxel, cisplatin, and camptothecin in HeLa and OVCAR-3 cells, the levels of M30 antigen were detected in the cells and their media by immunofluorescence staining and ELISA methods, respectively. RESULTS: The percentages of M30-fluoresein isothiocyanate (FITC) positive cells in HeLa and OVCAR-3 cells treated with paclitaxel, cisplatin, and camptothecin were 4.3% vs 18.1% vs 34.87% and 4.07% vs 18.6% vs 32.63%, 4.3% vs 17.87% vs 32.38% and 4.07% vs 16.83% vs 32%, and 4.3% vs 16.75% vs 31.3% and 4.07% vs 15.18% vs 29.9% in control, low dose, and hight dose groups, respectively (P<0.001). M30 antigen levels (U/L) measured in culture media of HeLa and OVCAR-3 cells treated with paclitaxel, cisplatin, and camptothecin were 53.03 vs 101.53 vs 355.59 and 86 vs 114.41 vs 412.04, 53.03 vs 79.84 vs 327.64 and 86 vs 125.44 vs 385.09, and 53.03 vs 88.41 vs 295.005 and 86 vs 108.42 vs 263.1 in control, low dose, and hight dose groups, respectively (P<0.001). CONCLUSION: Our results obtained in this preclinical study suggests that measurement of the levels of M30 antigen may help to predict the clinical responses and to select the effective anti-cancer agents in clinical settings, rapidly and quantitatively.

Fertility-sparing radical abdominal trachelectomy for cervical clear cell adenocarcinoma unrelated to diethylstilbestrol exposure : A case report / 대한산부인과학회지

Clear cell adenocarcinoma (CCAC) is a rare cancer that comprises less than 9% of the cervical adenocarcinoma cases. We experienced a case of fertility-sparing radical abdominal trachelectomy for cervical clear cell adenocarcinoma (CCAC). Thus, reported it. A 27 year old female was diagnosed with clinical stage Ib cervical CCAC. She had no history of maternal exposure to diethylstilbestrol and had negative PAP cytology and HPV tests. She was treated with neoadjuvant chemotherapy followed by radical abdominal trachelectomy. After 2 cycles of postoperative adjuvant chemotherapy, the lesion disappeared completely in an imaging study, and potential fertility was preserved. Radical abdominal trachelectomy with chemotherapy may be a valuable approach for treating stage Ib cervical CCAC in women that wish to preserve potential fertility.

Myxoid neurofibroma of the labia majora: A case report / 대한산부인과학회지

Myxoid neurofibroma is a benign tumor of perineural cell origin, which is demonstrated with a positive immunohistochemical stainig for S-100. The most common locations of the myxoid neurofibroma are face, shoulder, arm, periungual and foot. We experienced an unusual case of myxoid neurofibroma which originated from labia majora area then reported it. To our knowledge, this is the first time that a labia majora location is reported.

Myxoid neurofibroma of the labia majora: A case report / 대한산부인과학회지

Myxoid neurofibroma is a benign tumor of perineural cell origin, which is demonstrated with a positive immunohistochemical stainig for S-100. The most common locations of the myxoid neurofibroma are face, shoulder, arm, periungual and foot. We experienced an unusual case of myxoid neurofibroma which originated from labia majora area then reported it. To our knowledge, this is the first time that a labia majora location is reported.

Diagnosis of Cervical Neoplasia Using Immunohistochemical Staining of p16(INK4A) / 부인종양

OBJECTIVE: There is need for more objective diagnostic parameters to identify cervical dysplastic or neoplastic cells. So, we examined the p16(INK4A) expression in the cervical tissues to evaluate the value of p16(INK4A) as a diagnostic parameter. METHODS: We examined the p16(INK4A) expression by immunohistochemical staining in normal cervical tissues (n=3), preneoplastic lesions (n=6), carcinoma in situ (CIS, n=5), and invasive carcinomas (n=5) of the cervix, which were selected randomly by H and E staining from the archives of formalin-fixed and paraffin-embedded tissues and we also examined the status of human papillomavirus (HPV) infection in the same tissues. RESULTS: The positive rates of p16(INK4A) expression was significantly higher in all abnormal cervical tissues including subclinical papillomavirus infection (SPI), dysplasia, CIS, and invasive carcinoma than in normal cervical epithelium (p=0.001). Despite the strong expression of p16(INK4A) in the area of CIS, no expression of p16(INK4A) was observed in the area of normal epithelium in the vicinity of CIS. 11 cases among 19 cases of examined tissue samples were tested for HPV infection. Seven of them showed positivity for HPV DNA. CONCLUSION: We herein demonstrated that p16(INK4A) would be a sensitive and specific marker for the abnormal cervical cells in tissue sections. This approach will help to reduce interobserver variations in the histopathologic interpretation of cervical biopsy specimens.

Expression of mullerian inhibiting substance and its receptor in ovarian neoplsia / 대한산부인과학회지

Mullerian inhibiting substance (MIS) is a glycoprotein hormone produced by fetal Sertoli cells that causes regression of the Mullerian ducts in males during sexual differentiation. Cell lines derived from human ovarian epithelium and rodent Leydig cell tumors, which respond to MIS in growth inhibition assays and express the MIS type II receptors (MISR II). But the pathophysiological role of MIS in human ovarian neoplasia development has not yet been fully established. In order to understand its role in pathogenesis of ovarian neoplasia, the expression and localization of the MIS and MISR II were studied in 5 normal ovaries, 11 benign tumors, 9 borderline ovarian malignancies, 40 ovarian malignancies in paraffin embedded tissue and tissue microarrays by using immunohistochemical stain. The results were as follows; 1. The first staining for MIS and MISR II were detected in granulosa cells in primary follicles of normal ovary. Among the growing follicles, larger developing follicles stained more intensely than smaller follicles. 2. In benign ovarian tumors, 8 (72.73%) in MIS and 5 (45.45%) in MISR II out of 11 cases were stained. The intensity scores of staining were 1.18 in MIS and 0.64 in MISR II. 3. In borderline malignancies, 6 (66.67%) in MIS and 7 (77.78%) in MISR II out of 9 cases were stained. The intensity scores of staining were 0.89 in MIS and 1.22 in MISR II. 4. In ovarian malignancies, the expression of MIS and MISR II were 50% (9/18) and 50% (9/18) in epithelial, 92.30% (12/13) and 76.72% (10/13) in germ cell, and 88.9% (8/9) and 100% (9/9) in sex-cord stromal tumors. The intensity scores of MIS and MISR II expression were 0.72 and 0.72 in epithelial, 1.45 and 1.62 in germ cell, and 1.78 and 1.67 in sex-cord stromal tumors. 5. There was significant high expression of MIS and MISR II in non-epithelial (90.91%, 86.36%) than epithelial ovarian cancers (50%, 50%). The scores of expression intensity was also higher in non-elithelial cancers (MIS: 1.67 +/- 0.16 vs 0.72 +/- 0.20, p=0.003, MISR II: 1.64 +/- 0.20 vs 0.72 +/- 0.21, p=0.022). In conlusion, the expression of MIS and MISR II were not different according to the differentiation, but tissue type specific. The frequency of MIS and MISR II expression was higher in non-epithelial cancers, especially in sex-cord stromal tumors. The results of this experiment could be utilized as scientific basis of researches, furthermore clinical applications in diagnosis and treatment of non-epithelial ovarian malignancies.

Serum Levels and Expressions of Inhibin A and Inhibin B in the Ovaries of Perimenopausal Women / 대한산부인과학회잡지

OBJECTIVE: To understand the physiologic effects and secretion pattern of inhibin A and inhibin B during menstrual cycle and menopausal transition, inhibin A and inhibin B levels were measured. And to detect any changes in expression of inhibins in human ovary with age, we examined immunohistochemical staining of alpha, beta A, and beta B subunits of inhibin in ovarian tissues. This study was also designed to investigate whether or not inhibin is an early marker for menopausal transition. METHODS: Inhibin A and inhibin B levels were measured in 320 samples from normal reproductive women, in 60 from perimenopausal women, and in 20 from menopausal women by ELISA. And we examined the immunohistochemical staining of alpha, beta A, and beta B subunits of inhibin in ovarian tissues of 35 normal reproductive, 20 perimenopausal, and 5 menopausal women, respectively. RESULTS: In the normal reproductive women, inhibin A begins to increase in the late proliferative phase (16.53 +/- 1.57 pg/ml), reaches the peak in the mid-secretory phase (45.85 +/- 2.08 pg/ml), and subsequently decreases. Inhibin B begins to increase in the early proliferative phase (65.40 +/- 4.08 pg/ml), reaches the peak in the ovulatory phase (110.74 +/- 9.83 pg/ml), and thereafter declines rapidly. In the perimenopausal women, mean inhibin A serum concentration was 6.68 +/- 0.53 pg/ml during proliferative phase and 21.78 +/- 3.61 pg/ml during secretory phase, which were significantly lower than that of the same phase in the normal reproductive women (P<0.01). Mean inhibin B serum concentration was 52.16 +/- 7.46 pg/ml during proliferative phase and 22.41 +/- 6.73 pg/ml during secretory phase, which were significantly lower than that of the same phase in the normal reproductive women (P<0.01, P=0.025). In the menopausal women, both inhibin A and inhibin B were not detected. In the normal reproductive women, we observed strong immunostaining for alpha subunit in granulosa cells, theca cells, and corpus luteum. Immunostaining for beta A subunit was observed in corpus luteum, but not in growing follicles. Immunostaining for beta B subunit was observed in primary follicle, granulosa and theca cells of growing follicle, and mature follicle, but less strong than immunostaining for alpha subunit. No staining for beta B subunit was observed in the corpus luteum. In the perimenopausal women, immunostaining for inhibin subunits were observed in the same pattern as that of the normal reproductive women, but weaker. Stronger immunostaining was observed in theca cells than in granulosa cells. In the menopausal women, none of the immunostaining of inhibin subunits were observed. CONCLUSION: It is concluded that inhibin A is associated with the luteal function and inhibin B, the follicular function. The secretion of inhibins decreased rapidly in the perimenopausal transition period and were not detected in the menopausal period. Inhibin A and inhibin B are associated with the follicular maturation and development. It suggests that the inhibin A and inhibin B are good candidates as markers for perimenopausal transition.

Detection of p16(INK4A) in the Mixed Cell Populations of Normal Peripheral Blood Mononuclear Cells and Cervical Cancer Cell Lines / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Human papilloma viruses (HPVs) play a central role in the pathogenesis of neoplastic lesions of the uterine cervix. The viral oncoprotein HPV E6 degrades the p53 protein, and the HPV E7 protein inactivates pRB and increases the expression of the CDK inhibitor, p16(INK4A). We investigated the usefulness of p16(INK4A) as a biologic marker for the cervical dysplastic and neoplastic cells. MATERIALS AND METHODS: We examined the expression of p16(INK4A) and cytokeratin in a mixed population of normal peripheral blood mononuclear cells (PBMC) and the cervical cancer cell lines (HeLa, SiHa, and CasKi) using flow cytometry. RESULTS: The DNA indices of the HeLa, SiHa and CasKi cell lines were 1.89, 1.53 and 1.75, respectively, indicating that these cells are aneuploid cells. Furthermore, the positive rate of p16(INK4A) expression was 86.7% for the HeLa mixed population, 85.6% for the SiHa mixed population, and 92.2% for the CasKi mixed population. According to the FL3A vs FL3W histogram, electrical gating of the HeLa, SiHa and CasKi mixed populations showed the expression levels of both cytokeratin and p16(INK4A) to be identical, at 86.6%, 84.8% and 85.0%, respectively. These findings revealed that almost all cells selected through electrical gating were cervical cancer cells originating from the epithelium and which expressed cytokeratin and p16(INK4A). On the other hand, when each mixed population was electrically gated for normal PBMC, we found that the PBMCs expressed neither cytokeratin nor p16(INK4A). CONCLUSION: Using flow cytometry, we observed the enhanced expression of p16(INK4A) in cervical cancer cell lines. These RESULTS suggest the usefulness of p16(INK4A) for the selective detection of cervical dysplastic and cancer cells in the liquid-based samples, which are taken from the cervices and contaminated with blood and stromal cells.

Selective Protection of Normal Proliferationg Cells Against Anti-Neoplastic Chemotherapy by Stausporine Pretreatment / 대한산부인과학회잡지

OBJECTIVE: A major limiting factor in human cancer chemotherapy is toxicity in normal cells and tissues. Our goal was to determine whether normal proliferating cells could be protected from chemotherapeutic agents by taking advantage of the differential drug sensitivity of cell cycle G1 checkpoint in normal and cancer cells. METHODS: Normal peripheral blood mononuclear cells (PBMC) and ovarian cancer cell lines (OVCAR- 3 and SKOV-3) were initially treated with 10 nM of staurosporine for 48 hours. After removal of staurosporine contained media, both PBMC and ovarian cancer cells were treated with 20 nM of paclitaxel for 24 hours. Cells were then allowed to recover in drug-free medium for 4 days. The DNA contents and cell cycle changes were detected by FACScan flow cytometer in the cells harvested whenever the medium was changed. RESULTS: After pretreatment of ovarian cancer cell lines (OVCAR-3 and SKOV-3) with 10 nM of staurosporine followed by treatment with 20 nM of paclitaxel, both OVCAR-3 and SKOV-3 cells were selectively arrested in G2M phase of cell cycle by paclitaxel and they resumed their proliferative cycle to some extents after the drugs were removed and cultured with fresh media. However. pretreatment with 10 nM of staurosporine protected normal circulating PBMC that had been induced to proliferate in vitro with phytohemagglutinin from paclitaxel. Staurosporine-induced arrest of PBMC in G0/G1 phase was reversible, and arrested cells tolerated 10 nM of paclitaxel in culture. CONCLUSION: OVCAR-3 and SKOV-3 cancer cells can be targeted specifically with paclitaxel, following staurosporine-mediated, selective and reversible G0/G1 arrest in PBMC.

A Case of Steroid Cell Tumor, Not Otherwise Specified, with Massive Ascites / 대한산부인과학회잡지

Steroid cell tumors, not otherwise specified, are rare ovarian sex cord-stromal tumors with malignant potential. The majority of these tumors produce steroids (testosterone is the most common) and various virilizing symptoms such as hirsutism, temporal balding and amenorrhea, may appear in patients. Executive history taking, physical examinations, CT or sonography and hormonal studies are helpful in the diagnosis, but the confirmation of diagnosis is made via a staging operation and pathology. Treatments include operation, chemotherapy (i.e., BEP), GnRH agonist therapy and radiotherapy. We experienced a case of steroid cell tumor, not otherwise specified, with massive ascites, and elevated CA125, which we wish to report with a brief review of the literature.

Flow Cytometric Detection of Apoptosis and p53 Protein in OVCAR-3 and SKOV-3 Ovarian Cancer Cell Lines / 대한산부인과학회잡지

OBJECTIVE: Taxol (paclitaxel)-induced apoptosis was studied to understand their biological mechanism correlated with the expression of p53 in the SKOV-3 and OVCAR-3 ovarian cancer cell lines. MATERIALS AND METHODS: The SKOV-3 and OVCAR-3 cell lines were cultured in RPMI 1640 medium without taxol (control group) and with taxol for 24 h and 48 h (experimental group). After harvest, the cells were stained with annexin V-FITC (fluorescein isothiocyanate) and anti-cytokeratin antibodies (clone CAM5.2 and clone MNF116). They were washed and stained with p53 antibody. After then the secondary antibodies, i.e., FITC- or phycoerythrin (PE)-conjugated goat anti-mouse (GAM) immunoglobulin G (GAM IgG-FITC or GAM IgG-PE) were added in the cells and they were incubated in the dark. DNA of these cells were stained sequentially with propidium iodide (PI). Standard FACScan equipped with a 488 nm single laser was used for the analysis of these cells. RESULTS: Both of SKOV-3 and OVCAR-3 cell lines were arrested in the G2M phase after treatment of taxol, suggesting that these cells would eventually enter into the stage of cell death. Fractions of negative cytokeratin and positive annexin V and amount of sub-G0G1 fraction indicative of apototic fractions were lower in the SKOV-3 cell line compared with that in OVCAR-3 cell line, probably as a result of lower sensitivity of SKOV-3 cell line to the taxol. p53 expression were not detected in SKOV-3 cell line. On the basis of observed findings in SKOV-3 cell line and findings of high expressions of p53 regardless of taxol treatment, no increases in their expressions according to culturing time, and gradual increases in sub-G0G1 fractions and in fractions of negative cytokeratin and positive annexin V indicative of apoptosis in OVCAR-3 cell line, we concluded that the expression of p53 would not be associated with cell cycle changes and the arrest in the G2M pahse but associated with the appearance of apotosis. CONCLUSION: Our results suggest that flow cytometric detection of the apoptotic fractions would be an effective, fast, and accurate method for the chemosensitivity test in tumor cells before the administration of anti-cancer drugs in gynecologic cancer patients.

Detection of Apoptosis by the Stainings of Annexin V, Propidium Iodide and Cytokeratin in OVCAR-3 Ovarian Cancer Cell Line / 대한산부인과학회잡지

OBJECTIVE: This study was designed to estimate the chemosensitivity by a quantitative evaluation of the apoptotic cell fractions using flow cytometry. METHODS: The OVCAR-3 cells were exposed to 20 nM or 30 nM taxol for 0 (control), 24 and 48 hours, then removed the taxol contained media, and cultured further with fresh media without taxol. (1) Fluorescein isothiocyanate-conjugated Annexin V (Annexin V-FITC) and propidium iodide (PI) were added to one test tube to detect the apoptotic cell fractions and at the same time, PI was added to the other tube to stain the DNA. (2) Annexin V-FITC and cytokeratin (clone CAM5.2 and MNF116) were added to the test tube. They were fixed and permeabilized with 1% paraformaldehyde solution and 100% methanol. They were then incubated with phycoerythrin (PE)-conjugated goat anti-mouse immunoglobulin G (GAM IgG1-PE or GAM IgG2a-PE) and sequentially stained with PI for DNA. All the stained cells were analyzed by a FACScan flow cytometer. RESULTS: (1) After treatment of 20 nM or 30 nM of taxol, G2M arrest was observed in both of treatment groups, which increased with time. (2) The G0G1 sub-fraction indicative of apoptosis increased with increase of culturing time from 24 hrs to 48 hrs. (3) The early apoptotic cell fraction with positive annexin V-FITC and negative PI increased with increase of culturing time. (4) In cells stained sequentailly with annexin V-FITC, cytokeratin (CAM5.2 and MNF116), and PI after 30 nM taxol treatment, the early apoptotic cell fractions increased with increase of culturing time. However, their extent was somewhat lower than those observed by positive annexin V-FITC and negative PI in cells treated with 20 nM of taxol. CONCLUSION: The results of sequential stainings with annexin V-FITC, cytokeratin, and PI were consistent with the those of annexin V-FITC and PI with parallel DNA staining. Our results suggested that the level of apoptosis detected by flow cytometry could be a marker of chemosensitivity which could select the sensitive anti-cancer agents before administration to gynecologic cancer patients.

Pseudo-Meigs Syndrome Due to Subserosal Leiomyoma Diagnosed by MR Imaging: Case Report

We report a case of pseudo-Meigs syndrome due to a large subserosal leiomyoma in a patient with a high serum carcinogenic antigen 125 level. Initial clinical examination suggested disseminated malignant disease though the typical signal characteristics of leiomyoma, seen at MR imaging, led to the diagnosis of pseudo-Meigs syndrome.

The Changes of Cell Cycle Phase Fractions and Expression of p53 by the Treatment of Staurosporine in MCF-7 Cell Line / 대한산부인과학회잡지

OBJECTIVE: We investigated the effects on the cell cycle and p53 expression by the treatment of various concentrations of staurosporine to elucidate the molecular mechanism of staurosporine induced cell cycle arrest in MCF-7 cell line. METHODS: Various concentrations of staurosporine were treated in MCF-7 cells cultured with RPMI 1640 media. The harvested cells were fixed and permeabilized with 1% paraformaldehyde and absolute methanol. Then the cells were stained indirectly with anti-p53 primary antibody and FITC conjugated goat anti-mouse(GAM)-IgG secondary antibody. Sequentially DNA were stained with 0.1% RNase and PI solution. These stained cells were analyzed by the standard FACScan flow cytometer. The obtained results were analyzed further with WinList 3.0, and ModiFit LT software program. RESULTS: MCF-7 cells were arrested mostly in G1 phase of cell cycle at 5-10 nM of staurosporine, however, the cells were arrested in G2 phase at 20-100 nM of staurosporine. The expressions of p53 protein were higher in the MCF-7 cells treated with both concentrations of 10 nM and 100 nM of staurosporine compaired with the control cells. This suggests that the p53 may be involved in the mechanism of G1 and G2M arrest of cell cycle in MCF-7 cell. CONCLUSIONS: The points of arrest in cell cycle differred depending on the concentrations of staurosporine and these cell cycle arrests at G0G1 and G2M pahse were related with p53 protein expression. It suggested that these results could be extended to study for staurosporine to be usefull as a potential anti-tumor agent.

Multiparametric Flow Cytometry in Breast Cancer Cell Line (MCF-7) Stained with Fluorescein Isothiocyanate, Phycoerythrin, and Propidium Iodide / 대한암학회지

PURPOSE: Multiparametric flow cytometry is a powerful tool for analyzing the phenotypic, cell kinetic and ploidy heterogeneity of tumor cell populations. But there are major problems such as inaccurate results by the contribution of non-neoplastic cell contamination and the substantial spectral overlap of PI (propidium iodide) into PE (phycoery- thrin) fluorescent emissions on a standard flow cytometer. Recent studies suggested that the emission spectral overlap from PI into PE could be sufficiently compensated electrically and the cytokeratin, a marker for epithelial tumor cells, are successfully used in conjunction with DNA specific dye so as to obtain DNA profiles selectively for cytokeratin-positive tumor cells. The aim of this study was to investigate the feasibility that multiparametric analysis in heterogeneous cell populations of cell lines like solid tumors, which were stained triply with PE, fluorescein isothiocyanate FITC, and PI, can be done without any influences by the contaminated normal diploid cell populations and without spectral overlap between fluorochromes on a standard flow cytometer. MATERIALS AND METHODS: MCF-7 cell lines and heterogeneous cell populations mixed with MCF-7 cells and human peripheral blood lymphocytes were fixed with 1% paraformal- dehyde and permeabilized with 100% methanol. Cytokeratin was labeled with PE and some proliferat!on-associated markers were labeled with FITC, which were followed by DNA staining by PI. These triply stained cells were measured on a standard FACScan flow cytometer equipped with 488 nm single laser and those acquired data were analyzed with WinList 3.0 and ModFit LT software programs on personal computor. RESULTS: Coefficient of variation (CV) of GoG1> peak of MCF-7 cells alone was 4.3. GoG1, S, and G2M phase fractions were 44.9%, 45.9%, and 9.2% respectively. FITC, PE and PI fluorochromes could be detected without any interference between them. CVs of GoG1 peak of PBL and MCF-7 cells in those heterogeneous population were 2.3 and 4.2 respectively. The DNA index of MCF-7 cells was 1.7. MCF-7 cells expressed the cyto- keratin, PCNA, p53, c-erbB/2 and c-myc antigen and in contrast, PBL did not express cytokeratin. The cell cycle phase fractions and oncoprotein expressions could be detected separately in diploid PBL and aneuploid MCF-7 cells in the mixed cell population without any influences by each other. CONCLUSION: These results suggested that the cellular antigen expressions of the malignant cells can be analyzed selectively without influences of fluorescent signals from nonneo- plastic cells. The neoplastic tumor subpopulations are clearly identified on the basis of both ploidy status and antigen expressions. The positive cytokeratin expressions indicate that they were derived from the epithelium, providing objective evidence of the tissue of origin and more precise analysis of DNA contents, ploidy, and oncogene expressions selectively with possible correlation between them. Thus, this method offers new possibilities for multiparameter flow cytometric analysis in the heterogeneous solid tumor cell populations.

Comparison of the Clinical Outcomes Between Cisplatin-Chemoradiotherapy and Radiotherapy Alone in Patient with Cervical Cancer / 대한부인종양콜포스코피학회잡지

We compared the therapeutic effects of concomitant chemoradiotherapy (CRT) using cisplatin to single radiotherapy (RT) in uterine cervical cancer. 34 cases of non-operable uterine cervical cancer were reviewed retrospectively from Mar, 1993 to May, 1996 in St. Mary' s Hospital. The patients were randomly selected to compare the effects of both methods. 22 patients were included in CRT group and 12 patients in RT group. The results were as follows: 1. The decrease of tumor size was not significant (2.17 cm in CRT and 1.95 cm in RT) (p=0.61), but the number of responders of CRT group was larger than that of RT group significantly (p0.05) 3. The overall survival rate showed no difference between two groups (p>0.05). The disease-free survivals for 38 months were 17.02% in CRT and 11.36% in RT, but it was not significant (p>0.05). In conclusion, concomitant chemoradiotherapy showed better rate of response, but size of tumor decrease and tumor markers showed no difference. CRT might improve the overall survival and disease-free survival, although it was not significant in this study. The clinical significance of CRT remains to be determined in large randomized clinical trial.

One Case of Neuroblastoma arising in Mature Cystic Teratoma of the Ovary / 대한부인종양콜포스코피학회잡지

The incidence of malignant change of ovarian mature teratoma is 1~2%. The majority is squamous cell cancer, the others was adenocarcinoma. Neuroepithelial tissue was frequently detected in mature cystic teratoma, but their malignant change was extremely rare. Only, two cases of neuroblastoma of ovarian teratoma were reported in the world. We report one case of neuroblastoma arising in ovarian mature teratoma with a brief review. Our case is the third reported one in the world.

Simultaneous Three Color Detection of Surface Antigen (My 7), Intracellular Antigen (c-myc), and DNA Content using Single Laser Flow Cytometry / 대한산부인과학회잡지

Flow cytometry, a useful tool for measuring DNA content and cell differentiation as expressed by cell surface markers, is utilized to measure multiple antigens, especially surface antigen, intracellular oncoprotein, and DNA content, simultaneously. For this simultaneous detection, several methods off ixation and permeabilization have been used with limited values. In this study, 20 ng/ml of lysolecithin in 1% paraformaldehyde solution was utilized for fixation and permeabilization of cultured promyelocytic leukemic cells(HL 60). The cells were first stained with phycoerythrin (PE)-conjugated monoclonal antibody to the cell surface My 7 antigen and then were fixed and permeabilized with 20 ng/ml of lysolecithin in 1% partormaldehyde solution. After incubation, the fixed and permeabilized cells were stained with monoclonal antibody to intracellular c-myc antigen, which were followed by fluorescein isothiocyanate (FITC)-conjugated secondary antibody. The c-myc stained cells were finally stained for DNA content with 7-amino-actinomycin D(7-AAD). This procedure permits excellent staining for intracellular oncoproteins and preservation of surface antigens with relatively low cofficients of variation (CV) for the G0G1 peak of the DNA histograms and suggests that the sequential staining procedure of surface antigen, intracellular antigen, and DNA content will be extended for the study of correlations with cellular differentiation, expression of oncoproteins, and cell cycle analysis in the cells which are obtained from human malignant diseases using a 488 nm single laser flow cytometry.

Exression of p53, Proliferating Cell Nuclear Antigen(PCNA), c-myc, and Epidermal Growth Factor Receptor(EGFR) in Cervical Neoplasia / 대한산부인과학회잡지

No abstract available.