MspI restriction fragment length polymorphism at the glycoprotein hormone alpha-subunit locus. Association of certain genotypes with neoplasia.

A restriction fragment length polymorphism (RFLP) in the human glycoprotein hormone common alpha-subunit gene has been identified and partially characterized in normal lymphocytes and placentae, established tumor cell lines, and tumor biopsy samples. High molecular weight DNA was digested with the restriction endonuclease MspI, separated by electrophoresis in agarose gels, transferred to nylon membranes by the method of Southern, and hybridized to 32P-labeled human chorionic gonadotropin alpha-subunit cDNA. After autoradiography, bands were detected at 5.3, 3.3, 2.1, 1.6, 0.8 and 0.6 kbp. Presence of the 5.3, 3.3 and 0.6 kbp bands was invariant and uninformative. Patterns missing the 0.8 kbp band and both the 2.1 and 1.6 kbp bands are consistent with separate alleles that occur in placental and lymphocyte DNA with frequencies of 0.44 (15/34) and 0.06 (2/34), respectively. Presence of all three bands (2.1, 1.6 and 0.8 kbp) is indicative of heterozygosity, occurring at a frequency of 0.50 (17/34). Additional restriction patterns, not yet observed in DNA isolated from term placentae or circulating lymphocytes, were detected in DNA obtained from tumor cell lines and fresh tumor tissues at frequencies of 0.79 (15/19) and 0.59 (10/17), respectively. Thus, particular alpha-subunit genotypes are disproportionately represented in tumor-derived DNA, occurring at frequencies 10- to 13-times higher than would be predicted from their occurrence in normal tissue. Paired normal and tumor tissues from the same individual exhibited identical hybridization patterns, suggesting that this RFLP may be representative of a predisposition toward a variety of neoplasias rather than indicative of a change in DNA structure at or near this locus as a result of tumor development.

New protein and PCR markers RAK for diagnosis, prognosis and surgery guidance for breast cancer.

Breast cancer antigens RAK-p120, -p42, -p25 were detected in 100% of breast cancer cases tested (71 cases). Only 10% of adjacent tissue cases tested positive for all three cancer antigens, and 17.5% of the cases tested positive for two antigens only. Eighty-five percent of histologically normal breast tissue samples, isolated either from breast cancer patients or patients with advanced fibrocystic disease, tested RAK-negative, with the exception of low expression of p25, observed in some patients. Polymerase chain reaction (PCR) with HIV-1 gp 41-derived primers revealed cancer-associated DNA fragments of similar size (140 bp) as in HIV-1 genome. Fifty-four percent of cancer adjacent tissues, and 50% of malignancy-free breast tissue samples, tested PCR-negative. It is suggested that genetic predisposition to cancer may be associated with the presence of RAK genes, while expression of RAK antigens marks an already ongoing process of malignant changes.

Patient-controlled analgesia in obstetrics and gynecology.

Patient-controlled analgesia, which provides pain relief through self-administration of intravenous doses of opioids, is widely available and advocated as an effective analgesic modality. This report reviews published experiences with patient-controlled analgesia during labor and after cesarean delivery or major gynecologic surgery. Currently employed devices allow accurate record-keeping of drug use and permit patient mobility. No one device has been shown to be preferable. The form of administration most commonly described is infusion of morphine or meperidine on demand without combined continuous basal infusion. During labor, brief but progressively intensifying episodes of pain undermine the effectiveness of these devices when used intravenously. Compared with intramuscular administration of narcotics, patient-controlled analgesia after cesarean or major gynecologic surgery has been judged by patients to be more acceptable in achieving a balance between tolerable pain and sedation. Respiratory depression has been rare and is often attributable to misprogramming. Costs of these devices can be justified with frequent usage and are at least partially offset by more efficient use of nursing personnel for pain management.

Loss of an X chromosome in aggressive angiomyxoma of female soft parts: a case report.

Cytogenetic analysis of an aggressive angiomyxoma of the vulvar region of a 16-year-old female revealed loss of one X chromosome (45,X,-X) in eight of 20 metaphase cells analyzed. Fluorescence in situ hybridization (FISH) performed on disaggregated cells from paraffin embedded lesional tissue confirmed loss of an X chromosome in 31% of cells. Cytogenetic analysis performed on peripheral blood showed a normal chromosomal complement (46,XX). Thus, loss of one X chromosome appears to be confined to the neoplasm. This anomaly has not been previously described in aggressive angiomyxoma.

The intraoperative detection of ovarian adenocarcinoma using radiolabeled CC49 monoclonal antibody and a hand-held gamma-detecting probe.

A phase II study to evaluate the safety and efficacy of the 125I-radiolabeled anti-TAG-72 monoclonal antibody, CC49, as a component of a system for the intraoperative detection of occult ovarian cancer deposits was carried out at the University of Nebraska Medical Center. Patients entered into the study were to have surgery for evaluation of their disease status. The primary objective of this study was to determine the ability of a gamma-detecting probe (GDP), the Neoprobe 1000, to intraopertively localize sites of disease not identified by traditional surgical or radiographic evaluation. It was postulated that improved detection of cancer foci might allow for therapeutic excision or might result in an alteration of subsequent treatment. Ten patients were enrolled in the study between May 1993 and March 1994. Nine of the patients were undergoing second-look surgery after completing primary chemotherapy. The remaining patient was having surgery to assess possible cancer recurrence. All patients received an intravenous injection of 2 mCi/1 mg 125I-radiolabeled CC49 without complication. After a mean of 24.5 days, the patients' background radiation counts were deemed low enough for accurate intraoperative cancer localization, and surgery was performed. Any visibly or palpably abnormal areas were biopsied after being evaluated with the GDP. Any areas suspicious for malignancy by GDP evaluation were also biopsied. Two patients without evident disease by radiographic or surgical examination had histologically confirmed metastases localized by the GDP. Four patients had obvious disease at surgery which was variably confirmed by the GDP; two of these patients had baseline elevations in circulating TAG-72 antigen levels that may have affected binding of antibody to the tumor. This system of radioimmunoguided surgery was well tolerated and practical in its application, and it permitted disease detection that resulted in potentially beneficial changes in patient management.

Inhibition of cancer cell growth by internalized immuno-histone conjugates.

MAb NS 88 directed against breast cancer cells, which is internalized and translocated to the cell nucleus, was conjugated with histone and labeled with 125I. 125I-MAb-histone complexes (M(r) 250,000) were internalized by breast and cervical cancer cells and localized in the cytoplasm and chromatin. Electrophoretic analysis of the cells extracted from the conjugates revealed the same molecular weights of the cytoplasmic and chromatin complexes as those of the native conjugate. Nicotine (0.1%), which suppresses lysosomal degradation, stabilized the conjugates within the cell and prolonged the presence of nondegraded complexes inside the cytoplasm and chromatin from 1 day to at least 3 days. MAb-histone complexes, but not MAb alone, inhibited RNA synthesis and proliferation of cervical and breast cancer cells. A new application of internalized MAbs as the vehicles for protein inhibitors of transcription or replication is discussed.

Nalbuphine after major gynecologic surgery. Comparison of patient-controlled analgesia and intramuscular injections.

A randomized investigation compared the efficacy and safety of nalbuphine administered by two methods, a patient-controlled infuser system and intramuscular (IM) injections, after major gynecologic surgery. Forty-seven patients were randomly assigned to receive nalbuphine by either method. The 22 patients using the infuser were given a 2.0-mg, incremental dose with a 10-minute lock-out interval between doses. A similar group receiving 10-15 mg IM every three hours served as the control. Misprogramming, overdosage, depressed respiration and drug dependence were not encountered. Self-administration provided equally satisfactory sedation and more immediate pain relief without painful injections. Although patients with the infuser had the ability to self-administer more medication, they did not use higher doses of nalbuphine than did the IM group. The additional cost of the infuser system was offset by the satisfaction expressed by the patients and by the improved nursing efficiency. Nalbuphine administered with a patient-controlled infuser provided an effective balance between analgesia and sedation and offered advantages over IM injections.

Pap smear screening after hysterectomy.

Using the Pap smear is an effective means to screen for cervical cancer. This articles discusses the importance of performing a Pap smear of the vagina to detect vaginal cancer or its precursors.

Growth factor-mediated mechanisms of nicotine-dependent carcinogenesis.

The direct effect of nicotine on the expression of receptors for the tumor necrosis factor alpha (TNF alpha) and transforming growth factor beta (TGF beta) and the internalization, intracellular distribution and stability of these growth factors in cervical cancer cell line SiHa was studied. Nicotine at concentrations in the range 0.05-0.25% inhibited cell growth and it exerted strong apoptotic and cytolytic effects at concentrations above 0.5%. Nicotine at 0.1% stabilized and protected from degradation [125I]TNF alpha and [125I]TGF beta internalized by cervical cancer SiHa cell line. In the absence of nicotine, [125I]TNF alpha and [125I]TGF beta were internalized during the first hour of incubation and localized mainly in the cytoplasm and in smaller amounts in the nucleus. After 1 day of cell exposure to growth factors, only traces of radioactivity were detected inside the cells, which indicated that both growth factors were rapidly degraded. In the presence of nicotine, both [125I]TNF alpha and [125I]TGF beta were detected in high quantities and in a non-degraded form in the cytoplasm and chromatin during 5 days of incubation. In addition to the lack of growth factor degradation, the presence of nicotine induced a nuclear accumulation of growth factors, with up to 37% of the internalized [125I]TGF beta being in the chromatin. An increased intracellular accumulation of [125I]TNF alpha and [125I]TGF beta in cells exposed to nicotine occurred without changes in expression of the cell surface receptors. Nuclear accumulation of TGF beta was followed by increased RNA synthesis and a switch from the growth-promoting action of TGF beta to the strong growth inhibitory effect. Inhibition of the lysosomal degradation of growth factors by nicotine is discussed as a potential mechanism of tobacco-induced carcinogenesis.

Novel family of gynecologic cancer antigens detected by anti-HIV antibody.

OBJECTIVE: The reactivity of gynecologic cancer proteins with monoclonal antibody (MAb) directed against the human immunodeficiency virus I (HIV-I) was tested. METHODS: Cytoplasmic and nuclear proteins, extracted from a broad range of gynecologic cancers obtained during standard surgical procedures, were tested in Western blotting with MAb 5023 developed against the amino acid sequences 308-322 of the envelope protein gp120 of HIV-I. RESULTS: Three cell membrane proteins, M(r)l20,000 (p120), M(r)41,000 (p41), and M(r)24,000 (p24), and one chromatin protein, M(r)24,000 (p24), were detected by MAb 5023 in invasive, poorly differentiated cervical squamous-cell carcinoma; ovarian serous cystadenocarcinoma; poorly and well-differentiated endometrial carcinoma; vulvar squamous-cell carcinoma; and malignant mixed müllerian tumor. The same antigens were identified in cervical carcinoma cell line SiHa. Neither p120 nor p24 was recognized by other MAbs directed against the variable loop of gp120. Antigens p120 and p41 were undetectable in normal ovarian tissue and in biopsy samples of normal vaginal and rectal mucosa. Rectosigmoid cancer as well as colon carcinoma, lung carcinoma, and melanoma cell lines all tested negative. CONCLUSIONS: The identified antigens may represent either the products of human genes (proto-onc-ogenes) or, more likely, the products of an unknown virus specifically expressed in female cancer.

PDGF AA as mediator in nicotine-dependent carcinogenesis.

Effect of nicotine on PDGF AA and PDGF BB interaction with cervical cancer SiHa cells was tested. [125I]PDGF AA was internalized by cells and accumulated in the cytoplasm and nucleus (chromatin). In the absence of nicotine, maximal accumulation of [125I]PDGF AA inside the cells occurred after 1 day of incubation, which was followed by a progressive degradation of the growth factor during the next 2, 3 and 5 days of cell exposure. In the presence or 0.001 or 0.01% nicotine, accumulation of [125I]PDGF AA was slightly higher than in the absence of nicotine, and maximal accumulation occurred after 2 days or incubation. In the presence of 0.1 % nicotine, maximal accumulation occurred after 5 days of incubation and was 20 and 14 times higher in the cytoplasm and chromatin, respectively. Nicotine-postponed degradation and increased nuclear accumulation of PDGF AA resulted in activation of RNA synthesis and cell proliferation. PDGF BB, which was not internalized by cells did not respond to nicotine treatment. The proposed mechanism of nicotine-PDGF AA co-carcinogenesis may involve inhibition of growth factor degradation at the lysosomal level and an increased chromatin accumulation of the non-degraded PDGF.

Giant syncytia and virus-like particles in ovarian carcinoma cells isolated from ascites fluid.

Ovarian cancer cells were isolated from ascites fluid of 30 different patients diagnosed with cystadenocarcinoma of ovaries. Large colonies of malignant ASC cells were observed during the first week of cell growth in vitro. Colony formation was followed by fusion of cells and formation of large multinucleated and highly vacuolated syncytia. In contrast, cells isolated from the ascites fluid produced by patients with benign mucinous cystadenoma of ovaries did not form syncytia. Nonmalignant Brenner tumor cells, isolated from the ascites fluid, also did not form syncytia. Syncytia, but not the nonmalignant tumor cells, were immunofluorescence stained with an anti-human immunodeficiency virus type 1 (HIV-1) gp120 monoclonal antibody (MAb) and MAb RAK-BrI. Both MAbs recognized cancer-associated antigens RAK (for Rakowicz markers) p120, p42, and p25. Exposure of ASC cells to either the anti-HIV-1 gp120 MAb or MAb RAK-BrI inhibited syncytium formation. PCR with HIV-1 Env-derived primers revealed DNA sequences with over 90% homology to HIV-1 gp41 in syncytia and in ovarian cancer cells but not in normal ovary cells. Electron microscopic analysis revealed viral particles, hexagonal in shape (90 nm in diameter), with a dense central core surrounded by an inner translucent capsid and dense outer shell with projections. Negative staining detected membrane-covered particles (100 to 110 nm in diameter) in the cell culture medium. Incubation of normal breast cells with viral particles resulted in drastic morphological changes and syncytium formation by the transformed breast cells. The cytopathic effects of the identified virus resembled those of spumaviruses, which, in addition to their epitopic and genetic homology to HIV-1, might suggest a common phylogeny.

Immunodetection of squamous cell carcinoma-associated variant of nuclear protein.

A new monoclonal antibody, MAb C63.3, was developed by immunizing mice with the high molecular weight fraction of the cytoplasmic proteins from the cervical squamous cell carcinoma. In Western blotting, MAb C63.3 reacted with cytoplasmic and chromatin antigens expressed by cervical and vulvar squamous cell carcinomas and much weaker with normal cervical tissue and skin cells. Different molecular variants of C63.3 chromatin antigens were found in normal tissues (heavy variants, M(r) 65,000 and 59,000) compared to squamous cell carcinomas (light variant, M(r) 48,000). Diagnostic value of the cancer-associated M(r) 48,000 antigen is discussed. MAb C63.3 was internalized by cells and bound to the chromatin which suggests that antigen(s) C63.3 might represent a receptor for a yet unknown ligand (growth factor). It is suggested that the high molecular weight C63.3 antigens play a role in maintaining the nonmalignant phenotype.

Prognostic factors and long-term survival in endometrial adenocarcinoma with cervical involvement.

The clinical, surgical, and histopathologic data from 202 patients with endometrial adenocarcinoma with cervical involvement are presented. One hundred fifty-one (75%) had histopathologically confirmed cervical involvement at the time of their definitive surgery, while in 51 (25%) no cervical involvement was conclusively identified. The 5-year actuarial survival for patients with true surgical stage II endometrial carcinoma (N = 24) was 76%. Extrauterine disease was documented in 32% (27/84) of patients in which the primary treatment modality was surgical. The 5-year actuarial survival was 65% for all patients with clinical surgical stage II disease. There appeared to be a survival advantage for patients treated by radical surgery as compared with more conventional treatments, especially in patients with numerous high-risk factors. The subgroup of patients (N = 53) having tumor grossly involving the cervix had a 5-year survival of 48%. In this subgroup of patients, radical hysterectomy offered improved 5-year survival over more traditional forms of treatment, particularly compared with simple hysterectomy or combined treatment with radiation and surgery. Multivariate analysis positively correlated myometrial invasion, grade, uterine serosal involvement, lower uterine segment involvement, adnexal metastasis, pelvic metastasis, aortic node metastasis, and peritoneal cytology, with disease-free survival. Clinical and surgical findings correlated poorly; therefore, primary surgical evaluation is recommended when possible.