BACKGROUND: The HER-2/neu proto-oncogene (also known as c-ErbB-2) encodes a 185 kD transmembrane glycoprotein with intrinsic tyrosine kinase activity. Many studies revealed the correlation between the aberrant overexpression of HER-2/neu oncogene and poor prognosis of the malignant tumors such as breast, stomach, colon, lung cancers. But the significance of HER-2/neu oncogene overexpression as a prognostic factor in ovarian cancer remains controversial. OBJECTIVE: The aims of this study were to assess the prevalence of HER-2/neu oncogene amplification by polymerase chain reaction(PCR) and to evaluate the prognostic significance of HER-2/neu oncogene overexpression in terms of chemo-responsiveness and survival rate. MATERIALS AND METHODS: This study included 32 patients with advanced ovarian cancers(24 epithelial ovarian cancers, 2 Brenner tumors, 2 malignant mixed miillerian tumors, 2 granulosa cell tumors, 1 struma ovarii, 1 Krukenberg tumor). All patients had underwent staging laparotomy, and postoperative adjuvant chemotherapy with platinum-based combination chemotherapy. PCR was performed using tissues preserved in liquid nitrogen at the time of debulking operation. Overexpression of HER-2/neu oncogene was defined as being equal to or greater than 1.5 a.u. We analyzed whether the HER-2/neu overexpression correlated with chemoresponsiveness and 5-year survival rate(5-YSR). RESULT: HER-2/neu oncogene amplification was present in all of the ovarian cancers(32/32). Significant overexpression[gene copy number(GCN) > or =1.5 a.u.] was present in 13 of 32 ovarian cancers(41%) and 12 of 24 epithelial ovarian cancers (50%). The clinical response rate to chemotherapy in high copy group(GCN > or = 1.5 a.u.) was 67%(8/12) and that of low copy group(GCN<1.5 a.u.) was 92%(11/12)(p>0.05). Pathologic response rate to chemotherapy was 0%(0/5) and 50%(3/6), respectively(p>0.05). 5-YSR was 8% in high copy group and 25% in low copy group, but this difference was not statistically significant(p=0.17). CONCLUSION: HER-2/neu overexpression might be a poor prognostic factor, but this difference was not definitely elucidated by satistical analytsis in this study. Larger scaled prospective randomized study is needed to define the prognostidc significance of the HER-2/neu overezpression in ovarian cancer.
Breast ; Brenner Tumor ; Chemotherapy, Adjuvant ; Colon ; Drug Therapy ; Drug Therapy, Combination* ; Glycoproteins ; Granulosa Cell Tumor ; Humans ; Laparotomy ; Lung Neoplasms ; Nitrogen ; Oncogenes* ; Ovarian Neoplasms* ; Polymerase Chain Reaction ; Prevalence ; Prognosis ; Protein-Tyrosine Kinases ; Proto-Oncogenes ; Stomach ; Struma Ovarii ; Survival Rate

p73, a first p53 relative, has been identified at chromosome 1p36, a region that is deleted in variety of human cancers. This protein shares strong homology with p53 protein, suggesting functional similarities with p53. Indeed, p73 can activate p53 downstream genes inducing apoptosis or growth arrest in tumor cells lacking p53. This phenomenon leads us to investigate the function of p73 in ovarian cancer because aberrant p53 was very frequently found in this cancer. We hypothesize that DNA damaging agents trigger p53 dependent apoptotic pathway through p73 instead of p53 in ovarian cancer having aberrant p53. We selected SKOV3 ovarian cancer cell line having no p53 gene and treated this cell line with cisplatin. After the treatment, we examined the transcriptional level of p73 and p21. Moreover, to identify whether the status of p53 influence to the function of p73, we performed same experiment after inserting adenovirus mediated p53(Avp53) into cell line. We detected significantly increased transcripts of p73 whcn treated with cisplatin. But treated with Avp53 or combined treatment with cisplatin, the transcriptional levels were not changed. These data suggest that overexpression of p73 may be important to trigger apoptotic pathway when the p53 gene is lost, but not so important in cells having normal p53.
Adenoviridae ; Apoptosis ; Cell Line* ; Cisplatin ; DNA ; Genes, p53 ; Genes, Tumor Suppressor ; Humans ; Ovarian Neoplasms*

Twenty six cases of borderline ovarian tumor(BOT) were treated between Jan. 1985 and Dec. 1997 at the Department of Obstetrics and Gynecology, Hanyang University. The clinical records were reviewed for all patients including histopathology, clinical features, and follow-up. The frequency of BOT was 12%(26/214) of epithelial ovarian malignancies, and patients with these tumors tend to present at a younger age(36 yrs) than those with invasive carcinomas. In terms of histologic type, mucinous type(21/26: 81%) were more prevalent than serous tumor(5/21: 19%) in this study. The positive rate of CA 125 was 20% in serous, and the positive rate of CA 19-9 was 24% in mucinous tumor. The size of mucinous was larger than that of serous tumors(17.1 cm vs 9,3 cm). Almost all of these tumor categorized as early stage(stage I: 96%), however, only one patient with serous tumor had advanced stage of disease(stage III: 4%), Therefore BOT tend to be diagnosed as earlier than invasive carcinoma. About 2/3 of patients were treated as conservative surgery(unilateral salpingooophorectomy or enuclation). Postoperative adjuvant chemotherapy was not given about half of cases(13/26). Median follow-up was 43 months and recurrent case was found only one in serous tumor, All patients in this study are still alive and free of disease except one, 5-year survival rate was 100%. But large number of study and long-term follow-up are needed to make a decision to treat and manage of BOT.
Chemotherapy, Adjuvant ; Female ; Follow-Up Studies ; Gynecology ; Humans ; Mucins ; Obstetrics ; Ovary* ; Survival Rate

HPV E2 protein is known to act as a negative regulator of transcription and the disruption of E2 open reading frame by HPV integration can release suppression of E6 and E7 mRNA expression, resulting in uncontrolled cellular growth and malignant transformation by inactivating tumor suppressor gene products (p53, pRb). YY1 mutation of HPV URR has been suggested as one of indicator that explains development of cervical neoplasia by episomal type of HPV. To extend this hypothesis, we examined whether mutation(s) in specific sites of HPV URR is functionally related to the invasiveness of cervical neoplasia and the physical status of HPV DNA. The URR sequences were obtained by PCR amplification of HPV-16 genome from CIN and invasive cancer patients, cloned into pUC18 for sequencing, and into pBLCAT8+ for functional CAT assay. Our previous data classified HPV-infected patients into three groups: 3 cancer cases carrying episomal HPV DNA; 12 cancer cases carrying integrated HPV DNA; 12 CIN cases carrying episomal HPV DNA. The specific variants in HPV-16 URR were found in Korean women: GA transition at nt 7520 (100%, 27/27), AC transition at nt 7729 (70%; 19/27), and GA transition at nt 7841 (78%; 21/27). Selective mutations were observed at the YY1-binding sites of HPV-16 URR in the 3 patients with invasive cervical cancer, who having the episomal forms of HPV-16 DNA: AC transition at nt 7484 and GA transition at nt 7488 (YY1-binding site 2; from 7481 to 7489). Additionally, CT transition at nt 7785 (YY1-binding site 3; from 7781 to 7790) was found from 2 of 3 patients. No YY1 site mutations were detected in the 12 CIN patients and in the HPV-integrated invasive cancer patients. To determine whether these mutations have effect on the expression of HPV E6/E7 genes driven by URR, the transient transfection assay was employed using URR-CAT reporter plasmid. The relative activities of three URR mutants from episomal HPV-16 DNA of cervical cancers were 2- to 4-fold higher than that of HPV-16 URR prototype. In contrast, the URRs from integrated HPV-16 DNA in cervical cancer and from episomal HPV-16 DNA in CIN, where no mutation of the YY1-binding site was detected, showed similar levels of promoter activity to that of URR prototype.Our results support the hypothesis that the mutation at YY1 binding site is functionally related to the development of cervical neoplasia caused by episomal HPV-16 DNA in Korean cervical cancer patient. Thus, mutation in YY1 site of episomal HPV-16 URR may play a role of HPV integration in the progression of cervical cancer.
Animals ; Binding Sites ; Cats ; Clone Cells ; DNA* ; Female ; Genes, Tumor Suppressor ; Genome ; Human papillomavirus 16* ; Humans ; Open Reading Frames ; Plasmids ; Polymerase Chain Reaction ; RNA, Messenger ; Transfection ; Uterine Cervical Neoplasms

The cell cycle is the set of events that is responsible for the duplication of the cells. Recent studies indicate that cell cycle regulatory proteins, mainly the cyclins and cyclin-related genes, can be critical targets during oncogenesis. The genes and gene products normally control specific events in the cell cycles, particularly during the late G1 and early S phase and G2/M phase. A large body of date implicates cyclins in oncogenesis. The first evidence came from human cyclin A in oncogenesis. Cyclin A is expressed from the late G1 phase through the M-phase of the cell cycle. Cyclin A is known as positive regulator of cell cycle and participates in the tumorigenesis. Overexpression of cyclin A has been reported in several cancers. Ki-67 is a nuclear protein expressed during the cell cycle except in Go. The labeling index of Ki-67 in the tumor cell nuclei has been used as a good prognostic factor. In this study, we compared labeling index of cyclin A and Ki-67 to assess the feasibility between them with 30 cases of cervical intraepithelial neoplasia(CIN) and 20 cases of invasive squamous cell carcinoma(SCC)by immunohistochemistry. The results were as follow; 1. Cyclin A expressed in normal parabasal cells and their labeling index was 0.8+/-0.4%, while in CIN and invasive SCC 65.5+/-9.4% and 86.5+/-12.3% respectively. Ki-67 expressed in normal parabasal cells as 1.3+/-0.7% while in CIN and invasive SCC as 77.8+/-12.9% and 92.2+/-17.6% respectively. 2. In CIN, the expression of cyclin A increased according to the grades of the CIN as 32.5+/-5.7%, 75.8+/-9.0%, and 83.2+/-13.4% in CIN I, II and III respectively. The expression of the Ki-67 also increased according to the grades of the CIN as 51.8+/-9.8%, 87.9+/-11.3%, and 93.6+/-17.5% respectively in CIN I, II and III. 3. There was no differences of cyclin A and Ki-67 expressions according to the histologic types of invasive SCC. Above results suggests that the cyclin A labeling index could be used as a marker of tumor progression in the uterine cervical carcinoma as Ki-67.
Carcinogenesis ; Cell Cycle ; Cell Cycle Proteins ; Cell Nucleus ; Cyclin A* ; Cyclins* ; G1 Phase ; Humans ; Immunohistochemistry ; Nuclear Proteins ; S Phase

BACKGROUNDS: Human papillomavirus (HPV) infection is known as the major causative phenomenon in the development of cervical cancer. E6 and E7 proteins of oncogenic HPV types can play critical roles in immortalization and malignant transformation of cervical epithelial cells. From the previous epidemiologic data, long term use of oral contraceptives may be one of the risk factor for cervical cancer. PURPOSE: Investigation of estrogenic and anti-estrogenic effects on the proliferation of cervical cancer cells and gene expression of HPV under the regulation of HPV upstream regulatory region (URR) would help to explain the role of estradiol in HPV-associated pathogenesis of cervical cancer. METHODS: Cervical cancer cells (HeLa, CaSki and C33A) were cultured in vitro in the presence of 17 beta-estradiol or tamoxifen and the numbers of cells were directly counted to observe the growth stimulatory or suppressive effect of the treatment. The correlation between the growth regulatory effect and HPV E6/E7 gene expression was determined by reverse transcription-polymerase chain reaction (RT-PCR). The estrogenic effect on the promoter activity of HPV URR was further confirmed by transient co-transfection assays, which were conducted in C33A cells using the HPV-18 URR-CAT reporter plasmid. Supplemental effect of estrogen receptor on the URR promoter activity was also evaluated. To analyze the growth suppressive function at the higher concentration of estradiol or tamoxifen in HeLa cells, DNA fragmentation assay was performed. RESULTS: The proliferation of HeLa and CaSki cells was stimulated by estradiol at the concentration of physiological level (< or =1 X 10-6M), reaching maximal growth at 0.5 X 10-6M. At concentration of 0.1 X 10-6M, tamoxifen also stimulated the proliferation of HeLa and CaSki cells. In contrast to HPV-positive cervical cells, C33A cells were not influenced to cell proliferation by addition of estradiol at the physiological level, indicating that HPV might play role in growth stimulatory effect of estrogen or tamoxifen. Interestingly, the proliferation of HeLa cells was totally suppressed by estradiol and tamoxifen at the higher concentration (5 and 10 X 10-6M), whereas those of CaSki and C33A cellswere not responded and little suppressed at the concentration, respectively. The levels of HPV-18 E6 and E7 mRNA were significantly increased after treatment of 0.5 X 10-6M estradiol as determined by RT-PCR. Furthermore, transient transfection experiments using the URR-CAT reporter plasmid indicated that the increased expression of HPV E6/E7 genes was related with the growth stimulatory effect of estradiol and tamoxifen. In addition, co-transfection of estrogen receptor (ER) leads to an over 4-fold increase in CAT activity after treatment of estradiol or tamoxifen with 0.5 X 10-6M. When estradiol or tamoxifen was treated at the concentration over 5 X 10-6M for 96 hr, a typical DNA ladder, a indicative of apoptosis, was observed in HeLa cells. However, DNA ladder was not detected in C33A cells of which growth was some suppressed under same concentration of estradiol. CONCLUSION: At the physiological levels, estradiol stimulated the growth of HPV-positive cervical cancer cells and tamoxifen also did at the concentration of 0.1 X 10-6M. The increased expression of HPV E6/E7 at the physiologic levels appeared to be related with the growth stimulation of HPV-positive cervical cancer cells. Growth suppression observed at the higher concentration (5 and 10 X 10-6M) might be a indicative of apoptosis shown by DNA fragmentation assay in HeLa cells. Taken together, these data suggested that the concentration of estradiol (< or =1 X 10-6M) could be a risk-factor in HPV-mediated cerivcal carcinogenesis.
Animals ; Apoptosis ; Carcinogenesis ; Cats ; Cell Proliferation ; Contraceptives, Oral ; DNA ; DNA Fragmentation ; Epithelial Cells ; Estradiol ; Estrogens* ; Gene Expression ; HeLa Cells ; Human papillomavirus 18 ; Humans ; Plasmids ; Regulatory Sequences, Nucleic Acid ; Risk Factors ; RNA, Messenger ; Tamoxifen ; Transfection ; Uterine Cervical Neoplasms*

PURPOSE: To set up more simplified detection method for human papillomavirus (HPV) sequence polymorphism which could be used for the study of HPV-related pathogenesis, route of infection, and many other epidemiologic studies. MATERIALS AND METHODS: One hundred and thirteen cases of uterine cervical tissues containing HPV 16 DNA confirmed by polymerase chain reaction (PCR) from Korean women were subjected to investigate the URR gene mutations. PCR-amplified products were sequenced by the fluorescent dideoxy termination method and opposite strand sequencing was performed as required. The results obtained from sequencing were analysed to find the most hypervariable segment which contains the greatest number of variants and subjected to PCR-single strand conformation polymorphism (SSCP) analysis. RESULTS: Among the length of nucleotide position (np) from 7175 to 24, we found 60 sites (60/815=7.36%) of base substitutions. Segment from np 7743 to 24 was the most hypervariable and contain 20 kinds of variants. In this segment, C-to-T mutation at position 24, G-to-A at 7842, and T-to-C at 7781 were more frequently found than other sites. By comparing sequencing results with PCR-SSCP, we found 15 patterns distinguishable each other. All of the mobility shift occurred in the PCR-SSCP pattern could be accounted for by the base substitutions and nearly all of the DNA sequencing results observed were reflected as alterations in the PCR-SSCP patterns. CONCLUSIONS: We have assigned the hypervariable segment in one portion of URR which could be used as PCR-SSCP analysis. Identification of HPV polymorphism by PCR-SSCP is potentially useful for elucidating a number of epidemiologic questions such as the pathway of viral spread and so on.
DNA ; Female ; Human papillomavirus 16 ; Humans* ; Molecular Epidemiology ; Polymerase Chain Reaction ; Sequence Analysis, DNA

The squamous cell carcinoma of the uterine cervix is the most common malignant tumor among women in Korea. Since 1976, when a research result that human papillomavirus(HPV) infection played some role in the carcinogenesis of the uterine cervical carcinoma had been published, numerous reports supporting the result have been released. Among the types of the HPV, type 16 and type 18 are classified as high risk types because they are frequently found in cervical lesions with high grade dysplasia and invasive carcinomas. However, it is impossible to ascertain by host histologic or cellular changes which type of HPV is infected. The HPV genome is composed of six open reading frames (ORF' s) named as E1, E2, E4, E5, E6 and E7 in the early region. Among these oncoproteins HPV E6/E7 have been strongly suggested to be important in carcinogenesis. When HPV infects the epithelial cells, it promotes cellular proliferation. The cellular proliferation can be evaluated by immunohistochemistry with the antibodies for proliferating cell nuclear antigen(PCNA) and Ki-67. Because PCNA has long half-life, and can be detected 48 hours after completion of mitosis, an estimation of proliferating cells by PCNA could be inaccurate. The expression of Ki-67 antigen is more correct than PCNA for the evaluation of proliferation cells due to its short half-life and rapid degradation after completion of the mitosis. In this study, immunohistochemical staining was conducted to determine the rate of expression of HPV E6, E7 and Ki-67, correlation with relationship in carcinoma in situ and invasive uterine cervical cancer. Fifty cases of carcinoma of in situ(CIS) and invasive carcinoma were immunohistochemically stained, and the results obtained were as follows: 1) E6 protein of HPV type 16/18 was expressed in 5 of 14 cases(35.7%) of carcinoma in situ, in 3 of 7 cases of microinvasive carcinoma (42.8%) and in 12 of 20 cases of invasive carcinoma(60%) but there was no significant difference in expression between the carcinoma in situ and invasive cancer group (p=0.138). 2) E7 protein of HPV type 16/18 was expressed in 10 of 14 cases(71.4%) of carcinoma in situ, in 6 of 7 cases of microinvasive carcinoma (85.7%), and in 18 of 20 cases of invasive carcinoma(90%) but there was no significant difference in expression between the carcinoma in situ and invasive cancer group (p=0.138). 3) The cell fraction expressing Ki-67 was expressed in 5 of 14 cases(35.7%) of carcinoma in situ, in 5 of 7 cases of microinvasive carcinoma (71.4%), and in 18 of 20 cases of invasive carcinoma.(90%) The cell fraction expressing Ki-67 increased according to the progress of cervical cancer. 4) There was no statistical significance between HPV type 16/18 E6 protein and the cell fraction expressing Ki-67(p=0.09). 5) There was no statistical significance between HPV type 16/18 E7 protein and the cell fraction expressing Ki-67(p=0.17). The above results suggest that the cell fraction expressing Ki-67 increases according to the invasiveness of cervical cancer and E6/E7 protein seem to play a role in the progression of cervical cancer. However we were not able to reveal a relation between E6/E7 protein and the cell fraction expressing Ki-67 in progress of cervical carcinoma, and it is recommended that further studies should be undertaken.
Antibodies ; Carcinogenesis ; Carcinoma in Situ ; Carcinoma, Squamous Cell ; Cell Proliferation ; Cervix Uteri ; Epithelial Cells ; Female ; Genome ; Half-Life ; Humans* ; Immunohistochemistry ; Ki-67 Antigen ; Korea ; Mitosis ; Oncogene Proteins ; Open Reading Frames ; Proliferating Cell Nuclear Antigen ; Uterine Cervical Neoplasms*

BACKGROUND: Recurrence of ovarian carcinoma is difficult to diagnose by current diagnostic modalities. Positron emission tomography(PET) might be useful for detecting recurrence of ovarian carcinoma by producing images which reflect biochemical change of tissues rather than their physical characteristics using a positron emitting glucose analog, 2-[18F]-fluoro-2-deoxy-D-glucose(FDG), because glycolysis is increased in malignant tissue. OBJECTIVE: To determine if PET is sensitive for the detection and demonstration of recurrence of ovarian carcinoma and to compare detectability of PET to that of serum CA-125 and CT/MRI. MATERIALS & METHODS: Whole body PET scan was performed in thirty patients with epithelial ovarian carcinoma from March, 1996 to March, 1998 in Seoul National University Hospital. All patients received cytoreductive surgery and combination chemotherapy. The recurrence of ovarian carcinoma was declared by surgico-pathologic evidence or abnormal elevation of serum CA-125 level. CTI/Siemens scanner was used for PET. Ten mCi(370 MBq) of FDG was injected intravenously before whole body scan was obtained from the head to the lower leg. Regional transmission and emission scan was also obtained for areas of tumor. Serum CA-125 levels and CT/MRI findings by the time of PET were matched with PET results. Correlation analysis was performed between each diagnostic modalities and the recurrence of ovarian carcinoma. RESULTS: Ovarian carcinomas were recurred in twelve out of thirty patients. FDG PET detected the recurrence of ovarian carcinoma in ten patients(sensitivity = 66.7%; specificity = 86.7%; contingency coefficient, CC = 0.48; p = 0.003), and better than CA-125(sensitivity = 75.0%; specificity = 75.0%; CC = 0.28; p > 0.05) and CT/MRI(sensitivity = 38.5%; specificity = 57.1%; CC = 0.01; p > 0.05). CONCLUSION: FDG PET accurately predicted the recurrence of ovarian carcinoma and is a useful adjunctive diagnostic method. A prospective case-control study with more patients might be needed in the future.
Case-Control Studies ; Drug Therapy, Combination ; Electrons* ; Glucose ; Glycolysis ; Head ; Humans ; Leg ; Positron-Emission Tomography* ; Recurrence ; Sensitivity and Specificity ; Seoul ; Whole Body Imaging

Cellular oncogenes are expressed as an intrinsic part of the transformed or neoplastic phenotype. More than 60 of the known cellular oncogenes play a specific role in normal cellular development and differentiation. To examine the correlation between ras oncogene expression and the development of cervical cancer, this study investigated the reactivity of cervical intraepithelial neoplasia(CIN) and carcinoma of the uterine cervix by using anti-ras P21 mouse monoclonal antibody. The expression of ras oncogene significantly increased with the grade of malignancy from 11% in severe dysplasia, 30% in carcinoma in situ, 43% in microinvasive carcinoma, to 53% in invasive cancer. The expression of ras oncogene was not correlated with histologic type, tumor size, and nodal status of cervical cancer. It was concluded that expression of ras oncogene is related to early phase of carcinogenesis and tumor invasion of carcinoma of the uterine cervix.
Animals ; Carcinogenesis ; Carcinoma in Situ ; Cervix Uteri* ; Female ; Genes, ras* ; Mice ; Oncogenes ; Phenotype ; Uterine Cervical Neoplasms