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.

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 Mixed Germ Cell Tumor of Ovary: Complete Remission after Conservative Surgery and Bleomycin, Etoposide, Cisplatin (BEP) therapy / 대한산부인과학회잡지

Malignant germ cell tumor of ovary is very rare and one of the most malignant gynecologic neoplasms usually affecting younger women. we experienced a case of 19-year old female with mixed germ cell tumor of ovary which was composed of endodermal sinus tumor and dysgerminoma. we report this case with a brief review of the literature.

A Case Of Intraperitoneal Hemorrhage Of Intraligamentary Pregnancy / 대한산부인과학회잡지

A case of intraperitoneal hemorrhage of intraligamentary pregnancy is one of the most unusual accidents in obstetrics and carries a high degree of maternal and fetal morbity and mortality. The diagnosis of intraligamentary pregnancy is rarely ever made prior to laparotomy. We report a case of it with a brief review of literatures concerned.

A Case Of Squamous Cell Carcinoma Arising In Mature Cystic Teratoma Of Left Ovary / 대한산부인과학회잡지

Mature cystic teratoma of the ovary is the most common ovarian germ cell neoplasm representing 10-20% of all ovarian tumors. Malignant transformation rarely takes place, which is approximately 1-2% of all cases. The most common malignancy arising in mature cystic teratoma is squamous cell carcinoma. The prognosis of malignant transformation developing in mature cystic teratoma is poor and the management is not established due to the rarity of malignant degeneration. A case of stage Ia squamous cell carcinoma which developed in mature cystic teratoma of left ovary in a 43-year-old female is presented. The ovarian tumor was woman's fist-sized with intact capsule. Left oophorocystectomy was done. Above case with a brief review of literature is given.