Reference List

Notes: This is an initial descriptive report of two morphologically different cell lines (SKG-3a and SKG-3b) obtained from a human uterine cervical epidermoid carcinoma. These sublines were found to be derived from a moderately differentiated epidermoid carcinoma partially mixed with clear cell components. Both sublines had already been subcultivated more than 80 times since the initial separation. SKG-3a cells were much smaller in size and volume than 3b. The numbers of chromosomes were almost identical. Their cytology revealed anaplastic and pleomorphic features. SKG-3a was positive with periodic acid-Schiff stain and this changed to negative after an amylase digestive test, suggesting that the cells contained glycogen. An electron- microscopic examination confirmed the presence of many glycogen particles in 3a cytoplasm, but few in 3b. Tonofilaments and desmosomes were observed in both cytoplasms, suggesting epidermoid origin. In nude mice, SKG-3a produced clear cell epidermoid carcinoma with much glycogen, while 3b grew as a moderately differentiated epidermoid carcinoma with little glycogen. It is concluded that SKG-3a is derived from the clear cell epidermoid carcinoma and 3b is from the moderately differentiated epidermoid carcinoma. It is also clear that both were more heat sensitive at and over 39 degrees C than normal cells
Reference ID: 2567

Reference ID: 2699

Notes: Department of Obstetrics and Gynecology, Osaka Medical College, Takatsuki, Osaka 569-8686, Japan gyn017@pohosaka-medacjp
Abstract: Dihydropyrimidine dehydrogenase (DPD) and pyrimidine nucleoside phosphorylase (PyNPase) are the first and rate-limiting enzymes that regulate 5-fluorouracil (5-FU) metabolism, and tumoral DPD activity appears to be a promising predictor of 5-FU sensitivity. However, the regulatory mechanisms determining these enzyme activities have not been fully understood. We investigated the biological effects of epidermal growth factor (EGF) and transforming growth factor (TGF)-alpha on cell growth and tumoral DPD and PyNPase activities, and the subsequent effects on 5-FU sensitivity in uterine cervical carcinoma SKG-IIIb cells. The treatment of tumor cells with EGF or TGF-alpha resulted in a concentration-dependent increase in tumor cell growth and PyNPase activity, whereas tumoral DPD activity was inhibited. Their stimulatory effects on tumor cell growth correlated well with PyNPase activity, but were inversely related to DPD activity (P < 0.01). 5-FU sensitivity of tumor cells increased in the presence of EGF or TGF-alpha. These growth factors were shown to stimulate the first, rate-limiting enzyme activity in 5-FU anabolism and to inhibit that in 5-FU catabolism, leading to enhancement of the antiproliferative action of 5-FU at achievable therapeutic levels. The tumor environmental factors, EGF and TGF-alpha, may act as intrinsic regulators of DPD and PyNPase activities that affect the 5-FU sensitivity of individual tumors
Reference ID: 5252

Notes: Department of Obstetrics and Gynecology, Osaka Medical College, Osaka, Japan
Abstract: We investigated the effects of epidermal growth factor (EGF) and transforming growth factor-alpha (TGF-alpha) on migration, invasion and proteinase expression of gynecological cultured cancer cells (SKG-IIIb cervical squamous cell carcinoma, OMC-4 cervical adenocarcinoma, SNG-M endometrial adenocarcinoma and OMC-3 ovarian adenocarcinoma), and whether these growth factors affect thymidine phosphorylase/platelet-derived endothelial cell growth factor expression of tumor cells. Tumor cell migration along a gradient of substratum-bound fibronectin and invasion into reconstituted basement membrane were stimulated by 0.1-10 nM EGF and TGF-alpha in a concentration-dependent manner. The zymography of tumor-conditioned medium showed that the treatment of tumor cells with EGF and TGF-alpha resulted in the increase of type IV collagenases, stromelysin and urokinase-type plasminogen activator which was partly confirmed by immunoblot analysis. The expression of thymidine phosphorylase/platelet-derived endothelial cell growth factor which has angiogenic activity, was also upregulated by these growth factors. These results suggest that EGF and TGF-alpha act as positive regulators on the invasion process of gynecological tumor cells which may be associated with their stimulatory action on the motility of tumor cells, the expression of proteinases secreted by tumor cells and the angiogenic phenotype
Reference ID: 5253

Notes: Department of Obstetrics and Gynecology, Osaka Medical College, Japan
Abstract: The present study was undertaken to investigate the effects of the aminopeptidase inhibitor ubenimex (bestatin) on the invasive activity of cultured human uterine cervical carcinoma cells. The invasion of squamous cell carcinoma OMC-1 and SKG-IIIb cells, and adenocarcinoma OMC-4 and CAC-1 cells into reconstituted basement membrane (Matrigel) was inhibited by the presence of bestatin in a concentration-dependent manner. However, bestatin did not have any effect on tumor cell proliferation or migration. Immunoblot analysis of tumor-conditioned medium showed that the treatment of tumor cells with bestatin resulted in the reduction of the 72 kDa gelatinase level (gelatinase A, latent form) in the four cell lines examined, and the reduction of the 68 kDa gelatinase level (gelatinase A, active form) in SKG-IIIb cells. Bestatin inhibited hydrolyzing activities towards substrates of aminopeptidases in tumor cells, but did not directly inhibit gelatinase A. These results suggest that bestatin may inhibit the invasion of uterine cervical carcinoma cells possibly through the inhibitory mechanisms for production and activation of gelatinase A modulated by tumor aminopeptidases
Reference ID: 5254

Notes: Department of Obstetrics and Gynecology, Osaka Medical College, Japan
Abstract: We investigated the effects of epidermal growth factor (EGF) and transforming growth factor (TGF)-alpha on migration, invasion and matrix metalloproteinase (MMP) expression of uterine cervical-carcinoma SKG-IIIb cells, and whether these growth factors affect pyrimidine-nucleoside-phosphorylase (PyNPase) activity and 5'-deoxy-5-fluorouridine (5'-dFUrd) sensitivity of tumor cells. Tumor-cell migration along a gradient of substratum-bound fibronectin and invasion into reconstituted basement membrane were stimulated by 0.1 to 100 ng/ml of EGF and TGF-alpha in a concentration-dependent manner. The zymography of tumor-conditioned medium showed that the treatment of tumor cells with EGF and TGF-alpha resulted in an increase of the 92-kDa type-IV collagenase (MMP-9), which was confirmed by immunoblot analysis. These growth factors also up-regulated the expression of PyNPase activity of tumor cells and consequently enhanced the anti-proliferative action of 5'-dFUrd, a cytostatic that is biotransformed to 5-fluorouracil (5-FUra) by PyNPase. However, EGF and TGF-alpha did not have significant effects on the 5-FUra sensitivity of tumor cells. These results suggest that EGF and TGF-alpha, tumor environmental factors, simultaneously up-regulate the potential of uterine cervical-carcinoma cells to invade extracellular matrices and their PyNPase activity, which are subsequently associated with the specific action of 5'-dFUrd selectively killing tumor cells of gynecological origin with high invasive and metastatic potential
Reference ID: 5255

Notes: Department of Obstetrics and Gynecology, School of Medicine, Saitama Medical School
Abstract: The cytotoxicity including complement dependent cytotoxicity (CDC) and antibody-dependent cell-mediated cytotoxicity (ADCC) of two kinds of monoclonal antibodies (Mo-Ab), namely, MSN-1 which was obtained by means of an immunization procedure with intact SNG-II cells from human endometrial adenocarcinoma and HMST-1 obtained from hybridoma fused lymphocytes from lymph node with murine myeloma cells were studied by the 51Cr liberation test in vitro. 1) Against SNG-II, MSN-1 and HMST-1 showed significant CDC activity at 38.4 +/- 4.2% and 41.9 +/- 4.8% respectively, when a guinea pig complement was used. However, no significant CDC, action on SKG-IIIb was induced by MSN-1 or HMST-1. 2) Mo-Ab:MSN-1 and HMST-1 showed no significant liberation of 51Cr in relation to SKG-IIIb and SNG-II when using peripheral blood lymphocyte (PBL) separated by centrifugation gradient as the effector cells, and then the cytotoxicity of PBL alone against K-562 cell, SNG-II and SKG-IIIb was 40%, 9% and 2% respectively. This suggested that neither Mo-Ab had any ADCC activity and that SKG-IIIb and SNG-II cells were NK resistant cell-lines
Reference ID: 5256

Notes: Department of Obstetrics and Gynecology, Saga Medical School, Japan
Abstract: The cellular changes in a human cervical squamous-cell-cancer cell line (SKG-IIIb) were investigated after treatment with human recombinant interferon gamma and human fibroblast interferon beta. Cytoplasmic changes characterized by elongation, vacuolization and blurring of cytoplasmic borders appeared relatively early in both treatment groups. After two days of treatment with interferon gamma, the cells began to aggregate and to form pearl-like structures in the center of these aggregates; frequently seen were isolated cells with an eosinophilic cytoplasm and a pyknotic nucleus, similar to keratinizing cells. After six days of treatment with interferon beta, on the other hand, more than 20% of the cells began to show nuclear changes, including remarkable pleomorphism, multilobulation and multinucleation. The differences in biologic activities between the two interferons are discussed on the basis of these observations
Reference ID: 5257

Abstract: The E6 and E7 regions of human papillomavirus (HPV) type 16 were present in the DNA samples from cervical cancer cell lines, SKG-IIIa and SKG-IIIb, and those from cervical cancer tissues of three different patients. T601 cells, an NIH3T3 transformant obtained by transfection of DNA from a surgical specimen of a cervical cancer, also contained the E6 and E7 regions. The E6 region of HPV type 16 was expressed as mRNA in SKG-IIIa, SKG-IIIb and T601 cells. The E6 and E7 regions of HPV type 18 were present in the DNA samples from cervical cancer cell lines, SKG-I and SKG-II, and those from cervical cancer tissues of two different patients. SKG-I and SKG-II cells expressed the E6 region of HPV type 18 as mRNAs. These results strongly suggest that the E6 and E7 regions or the sequence surrounding these regions are important for maintaining malignant phenotype of cervical cancer cells
Reference ID: 5258

Abstract: Five cell lines, SKG-I, SKG-II, SKG-IIIb, QG-U and QG-H derived from cervical carcinomas of Japanese patients, were examined for the presence of human papillomavirus (HPV) DNA and the expression of viral mRNA. The DNA of HPV type 16 was shown to be linked covalently with SKG-IIb, QG-U and QG-H cell DNA, and HPV 18 DNA with SKG-I and SKG-II cell DNA. Although different regions of the HPV genome were integrated in these cell lines, the non-coding region and an early region including the E6 and E7 open reading frames (ORFs) were conserved in all cell lines. The complete genome of HPV 16 was found in QG-H cells by digestion of the DNA with a single-cut restriction enzyme. The other early region ORFs E1, E2, E4 and E5 were interrupted by flanking host cell DNA, suggesting that the integration into host cell DNA occurs preferentially in this region. HPV-specific mRNA species were detected in all five cell lines. In the three cell lines containing the HPV 16 genome, mRNAs hybridized with the early region of the genome, covering the entire E6 and E7 ORFs and a minor part of the E1 ORF, although the amount and size of the major mRNAs varied in these cell lines. These mRNAs did not hybridize with the late region of the HPV genome containing the L1 and L2 ORFs. In SKG-II, SKG-IIIb and QG-H cells we also detected c-myc and c-Ha-ras mRNA expression at about nine times the level of that in normal cells
Reference ID: 5259

Abstract: The anti-cancer effect of Etoposide was tested in in vitro in cultures of cells taken from a squamous cell carcinoma of the uterine cervix (SKG-IIIb). Growth inhibition was tested by the regrowth assay method, inhibition of DNA synthesis by the uptake of 3H-thymidine, and morphological changes by the method of Limburg et al. The concentrations of Etoposide used were similar to the blood levels recommended for clinical use. The regrowth assay showed that the effective concentration of Etoposide for 50% cell kill was 7.5 micrograms/ml in 2-hour and 1.0 microgram/ml in 24-hour cultures. The 3H-thymidine uptake test showed that a concentration of 13 micrograms/ml for 2 hours or of 3.5 micrograms/ml for 24 hours resulted in 50% inhibition of DNA synthesis. Morphological changes were much greater in the 2-hour cultures at the higher concentration of Etoposide than in the 24-hour cultures at the lower concentration. Investigation of the drug sensitivity by cell kinetics disclosed prolongation of the cell cycle occurring after 96 hours at 1.0 microgram/ml and inhibition of cell cycle progression occurring after 24 hours at 10.0 micrograms/ml and after 4 or 8 hours at 50.0 mu/ml. Thus, the anti-cancer effect of Etoposide on SKG-IIIb depends on both the concentration and exposure time and is related to its ability to inhibit cell growth and DNA synthesis and to cause morphological changes in the cancer cells
Reference ID: 5260

Abstract: Various types of human gynecological cultured tumor cells were tested for the sensitivity to Peplomycin (PEP), an effective antitumor antibiotic for squamous cell carcinomas, by the regrowth assay method together with morphological observation. Bleomycin-hydrolase activity of these cell lines was also compared in cell-free extracts by assaying the conversion of Bleomycin into its deamidated from (HPLC method). SKG-I, SKG-II, SKG-IIIb cells derived from squamous cell carcinoma of the cervix and RKN cells derived from myosarcoma of the ovary were much more sensitive to PEP than other cell lines. PEP was found to be mainly a time-dependent drug, but also concentration dependent. The effect of PEP on cell morphology was characterized by the appearance of enlarged cells and swelling nuclei. The specific activities of Bleomycin-hydrolase in SKG-I, SKG-II, SKG-IIIb cells were shown to be relatively lower than that in other cell lines. These results suggested that cervical squamous carcinoma cells and ovarian myosarcoma cells were sensitive to PEP and Bleomycin-hydrolase activity was one of factors which decided the PEP sensitivity of human cultured tumor cells
Reference ID: 5261

Abstract: The production of Regan isoenzyme (heat-stable, L-phenylalanine-sensitive term-placental alkaline phosphatase), human chorionic gonadotropin beta-subunit, and pregnancy-specific beta 1-glycoprotein by newly characterized human uterine cervical cancer cell lines, SKG-IIIa and SKG-IIIb, is reported. These cell lines were derived from a moderately differentiated epidermoid cancer partially mixed with epidermoid clear-cell components. At the end of the first 4 months in culture 2 sublines with different morphologies were identified. In nude mice, SKG-IIIa produce clear-cell epidermoid cancer with much glycogen, while SKG-IIIb grew as a moderately differentiated epidermoid cancer rich in tonofilaments. The presence of Regan isoenzyme was established by biochemistry, enzyme cytochemistry, immunocytochemistry, and immunoelectrophoresis. However, the copresence of small amounts of early placental alkaline phosphatase was also demonstrated. The alkaline phosphatase specific activities of SKG-IIIa cells and SKG-IIIb cells were 3.7 and 1.4 nmol per mg protein per min, respectively. The existence was proven by radioimmunoassay of human chorionic gonadotropin beta-subunit (SKG-IIIa, 5.0 mlU/mg protein; SKG-IIIb, 4.4 mlU/mg protein), pregnancy-specific beta 1-glycoprotein (SKG-IIIa, 0.7 ng/mg protein) in the culture media as a tumor cell product. The described cell lines may serve as a more representative model system for studies of regulation of oncodevelopmental genes in gynecological tumors in general and in epidermoid cervical cancer in particular
Reference ID: 5262