MIR-142-5p and miR-9 may be involved in squamous lung cancer by regulating cell cycle related genes.

OBJECTIVE: To identify key genes associated with squamous lung cancer (SLC) through analyzing gene expression data with bioinformatic tools, which could be potential biomarkers for diagnosis and treatment. MATERIALS AND METHODS: Gene expression data set GSE3268 was downloaded from Gene Expression Omnibus, including 5 SLC samples and 5 healthy controls. Data pre-treatment and differential analysis were performed with packages of R. Cluster analysis was done based on gene expression values to globally present the difference between the two states. Differentially expressed genes (DEGs) were divided into up-regulated and down-regulated genes, and then underwent functional enrichment analysis with DAVID tools. WebGestalt was used to retrieve microRNAs for the DEGs and then a regulatory network was constructed. GENECODIS was selected for functional annotation for all the genes in the network. RESULTS: A total of 537 DEGs were obtained. Functional enrichment analysis revealed that cell cycle was significantly enriched in up-regulated genes. Besides, two microRNAs (miRNAs), MIR-142-5p and miR-9, were retrieved, which were potential tools to regulate the expression of key genes. CONCLUSION: These DEGs may be involved in pathogenesis of SLC and some of them could be potential biomarkers. Besides, MIR-142-5p and miR-9 may be utilized to treat SLC as they could modulate cell cycle.

Reduction of estrogen receptor concentration in MCF-7 human breast carcinoma cells following exposure to chemotherapeutic drugs.

To study the effects of commonly used chemotherapeutic drugs on estrogen receptor (ER) of human breast cancer, we investigated the specific binding of [3H]estradiol within intact MCF-7 human breast cancer cells after 1 to 4 hr of exposure to methotrexate (0.5 to 50 micrograms/ml), 5-fluorouracil, and vincristine in serum- and hormone-free medium. Intracellular [3H]estradiol binding was either slightly increased (methotrexate and 5-fluorouracil) or not changed (vincristine) during the first 2 hr of drug exposures at 37 degrees, but slightly decreased at the third hr. After 4 hr of drug treatments, [3H]estradiol binding within MCF-7 cells was reduced by 30 to 70%; the response was dose dependent. Most (80 to 83%) of the intracellularly bound [3H]estradiol was found within the nuclei, and the drug-induced reduction of ER was reflected by a depleted nuclear uptake of [3H]estradiol. The Scatchard plot showed a large decrease of receptor number per cell with no apparent alteration in the binding affinity. The reduction of ER was reversible; regeneration of receptors to the control level occurred at either 4 hr (methotrexate and 5-fluorouracil) or 8 hr (vincristine) after removal of these drugs. The restoration was followed by an increase of ER beyond the control level. The dose-dependent depletion of ER by these cytotoxic drugs was also detectable in a second ER-positive cell line, MDA-MB-134. These data indicate that the cytotoxic drugs may cause a dose-dependent, reversible depletion of ER in human breast cancer, and the effect seems to be due to inhibition of receptor synthesis rather than inhibition of the binding of estradiol to its receptors.

Characterization of an inhibitor for luteinizing hormone receptor site binding.

An inhibitor for luteinizing hormone (LH) receptor site binding has been partially purified from aqueous extracts of ovaries from pseudo-pregnant or pregnant rats. The LH receptor binding inhibitor (LH-RBI) was heat-resistant, partially inactivated by trypsin or pronase digestion, and had a molecular weight of approximately 3800 daltons. The LH-RBI was not found in the ovary of mature non-pregnant rats or immature rats, nor was it found in testis or liver extracts. The LH-RBI strongly inhibited the in vitro binding of 125I-labeled ovine LH to ovarian LH receptors but did not inhibit 125I-labeled ovine prolactin binding to ovarian PRL receptors.

Insulin-like growth factor-II (IGF-II) messenger ribonucleic acid is expressed in steroidogenic cells of the developing ovine adrenal gland: evidence of an autocrine/paracrine role for IGF-II.

Insulin-like growth factors (IGFs) are potent mitogenic and differentiation-promoting factors that regulate the growth and development of many fetal tissues. Their role in the development of the adrenal gland and activation of its function is not known. The latter is crucial in providing the stimulus for the maturation of various fetal organs and determines the onset of parturition in sheep. To examine the hypothesis that IGFs are important autocrine/paracrine regulators of fetal adrenal development in vivo, we localized IGF-I and IGF-II mRNAs and peptides in the adrenal glands of developing sheep fetuses and correlated the cellular distribution with localization of 3 beta-hydroxysteroid dehydrogenase, tyrosine hydroxylase, and phenylethanolamine-N-methyltransferase enzymes by immunohistochemistry. Adrenal glands from 60- to 75-day-old (n = 4), 100- to 110-day-old (n = 4), 120- to 130-day-old (n = 4), and 145- to 147-day-old (term; n = 4) fetal sheep and 1- to 4-day-old newborn lambs (n = 4) were dissected and either snap-frozen or fixed. Total RNAs were subjected to Northern analysis using ovine IGF-I and IGF-II cDNA probes. Seven IGF-II transcripts of 1.2-6.0 kilobases (kb) were identified in the adrenal glands of fetuses at all gestational ages, and in the newborn. By densitometry, the abundance of IGF-II mRNA was highest in the fetal adrenal gland at 60 days, decreased slightly between 60 and 100 days, remained relatively constant until term, and decreased significantly after birth. At all gestational ages, IGF-II mRNA was detectable in significantly greater abundance than IGF-I mRNA. IGF-I and IGF-II mRNAs were localized by in situ hybridization using 35S-labeled anti-sense cRNA probes, and the peptides by immunohistochemistry using specific antisera. Low levels of IGF-I mRNA were detected in the zona fasciculata, but not in the zona glomerulosa. There was strong hybridization of the IGF-II cRNA to the zona glomerulosa and fasciculata and to the capsule. The hybridization signal was greater in the zona fasciculata than in the zona glomerulosa. IGF-II mRNA was also detected in groups of cells within the medulla. Localization of IGF-II mRNA by in situ hybridization correlated well with the distribution of IGF-II immunoreactivity and with 3 beta-hydroxysteroid dehydrogenase-positive cells in the cortex and in groups of cells within the medulla.(ABSTRACT TRUNCATED AT 400 WORDS)

Quantification of opioid-binding sites in the ewe hypothalamus.

Opioid-binding sites were quantified in the ewe hypothalamus using [3H]diprenorphine ([3H]DIP) as the radioligand. [3H]DIP binding to hypothalamic membrane preparations was stereospecific, saturable with respect to [3H]DIP concentration, and linear with hypothalamic membrane protein content. Scatchard analysis revealed a single class of binding sites. There were no significant differences in binding site concentration or binding affinity in hypothalami from intact ewes during the breeding and non-breeding seasons, or from long-term ovariectomized ewes with and without oestradiol treatment during the breeding season. Thus, whilst ovarian steroid hormones are known to modify LH responses to opioids and their antagonists in the ewe in vivo, they do not appear to do this by modulating the numbers of hypothalamic opioid-binding sites.

Controls of corticotrophin-releasing factor output by hypothalamic tissue from fetal sheep in vitro.

Corticotrophin-releasing factor (CRF) is thought to be an important physiological regulator of the pituitary-adrenal axis in fetal sheep and, as such, plays a fundamental role in the initiation of parturition in this species. However, little is known of the controls of CRF secretion from the fetal hypothalamus. We looked for the presence of CRF in fetal hypothalami, and examined whether the hypothalamic CRF concentration or molecular species changed in relation to gestational age. We established an in-vitro perifusion system to examine the release of CRF from perifused hypothalami taken from fetuses at day 100 and day 140 of pregnancy, under basal conditions and in response to potassium depolarization and/or dexamethasone administration. Immunoreactive CRF was present in fetal hypothalami as early as day 100 (2.42 +/- 0.99 (S.E.M.) micrograms/g protein, n = 9) and in similar concentrations at day 140 (2.31 +/- 0.69 micrograms/g protein, n = 9). There was a significant (P less than 0.05) increase in hypothalamic CRF content to 14.79 +/- 4.09 micrograms/g protein (n = 16) between day 122 and day 135 of gestation. Using Sephadex G-75 chromatography, hypothalamic extracts at day 100, days 122-135 and day 140 eluted with a single peak of immunoreactivity which corresponded to synthetic ovine CRF(1-41). The basal release of CRF from perifused hypothalami at day 140 (76.6 +/- 10.4 pg/fraction, n = 8) was significantly (P less than 0.05) greater than at day 100 (50.1 +/- 10.2 pg/fraction, n = 11). Dexamethasone significantly inhibited basal CRF release at day 140 of gestation but not at day 100.(ABSTRACT TRUNCATED AT 250 WORDS)

Hypercalcemia in patients with known malignent disease.

In 82 patients, a preoperative diagnosis of primary hyperparathyroidism has been established by means of transfemoral neck vein catheterization and measurement of serum immunoreactive parathyroid hormone (iPTH). Twenty-five of these patients have had cancer in other parts of the body but with no evidence of recurrence or metastasis. One patient had carcinoma of the colon with metastases, and four were members of families with multiple endocrine adenomatosis (MEA, Types I and II). In six other hypercalcemic patients, high levels of iPTH were found also in the effluent blood from cancer sites other than the parathyroid gland, secondary to ectopic hormone production or pseudohyperparathyroidism. In addition, a high serum level of iPTH was found in the superior vena cava of a seventh patient who had carcinoma of the breast but no clinical or radiological signs of recurrence or metastasis with the exception of an enlarged liver. This iPTH finding was interpreted as being, probably, the result of parathyroid adenoma in either the neck or the mediastinum. At the time of operation, a transcervical mediastinal search was made. Four normal cervical parathyroid glands were found; three were removed. Hypercalcemia persisted after operation, and the patient died. At postmortem examination, microscopic study revealed that the disease had metastasized to lungs and hilar lymph nodes. There was massive metastasis in the liver; the liver contained a large amount of iPTH. The results of these investigations suggest that (1) venous catheterization of the neck veins and the effluent blood from extraparathyroid tumors aid in identifying and localizing iPTH production; (2) primary benign hyperparathyroidism is not uncommon in patients with cancer, and its co-existence must be recognized; (3) high serum iPTH level in the superior vena cava may be found in patients with metastatic or primary cancer of the thoracic cavity; and (4) hyperparathyroidism may be the first hint of a familial multiple endocrine syndrome.

Enhancement of hormonal synchronization and 5-fluorouracil cytotoxicity in breast cancer cells by low concentration of thymidine.

A method is described that applies a low concentration of thymidine to achieve maximal hormonal synchronization in cultures of an established human breast cancer cell line, MCF-7. The method consists of sequential treatments with tamoxifen, estradiol, and thymidine. The increase (3.8 fold) in the S-phase fraction achieved by this method is higher than that (2.6 fold) achieved by a solely published method using tamoxifen block plus estradiol rescue. The MCF-7 cells, which have an enhanced labeling index (S-phase fraction) after being treated with the synchronizing method, have a markedly increased sensitivity to an S-phase-specific cytotoxic agent, 5-fluorouracil. This method may have potentially important implications for clinical chemotherapeutic strategy in the treatment of breast cancer.

Lethal efficacy of doxorubicin on human medullary thyroid carcinoma cells in vitro.

Cytotoxicity of doxorubicin was evaluated on a cultured human thyroid carcinoma (MTC) cell line, TT, by the colony-formation technique. The concentration-dependent survival curve showed a biphasic exponential pattern. Doxorubicin in concentrations of 5 X 10(-9) to 3 X 10 (-5) M produced a 15% to 71% cell kill after 1 hr of treatment. Mean lethal concentrations, 0.6 and 59 micrograms/ml, were considerably higher than those reported for cells of other tumors. Prolonged continuous treatment with a single concentration (1 X 10(-8) M) resulted in a cell kill of only 39% by 20 hr, and no further improvement was achieved with extended treatment of up to 48 hr. That doxorubicin activity was not enhanced by prolonged treatment was shown by controls not to be due to inactivation of the drug. Our results suggest that TT cells are somewhat resistant to doxorubicin and that acute administration of larger doses rather than continuous infusion of small doses should perhaps be considered when doxorubicin is used in patients with metastatic MTC.

Intrinsic drug resistance in a human medullary thyroid carcinoma cell line: association with overexpression of mdrl gene and low proliferation fraction.

Medullary thyroid carcinoma (MTC) is a cancer that is relatively insensitive to clinical chemotherapy. Our previous studies have demonstrated that an established human MTC cell line, TT, seems to possess an intrinsic resistance phenotype when tested for its chemosensitivity to multiple antineoplastic agents. We now report our investigation on the potential mechanisms responsible for the chemoresistance of TT cells. Northern analysis showed an increased level of multidrug resistance gene (mdrl) mRNA in TT and in an inherently drug-resistant colon carcinoma cell line, LoVo, when compared with CEM, a drug-sensitive leukemic lymphoblastic cell line; the latter two cell lines were included here as a control. Verapamil (10 microM) partially reversed resistance to doxorubicin in TT and LoVo cells, but had no effect on doxorubicin cytotoxicity to CEM cells. Expression of glutathione-S-transferase-pi (GST pi) gene was undetectable in TT, whereas, under similar conditions, GST pi mRNA was detectable in LoVo. Growth kinetics studies revealed that doubling times of the 3 cell lines in exponential growth were 95, 37, and 24 h for TT, LoVo and CEM, respectively. Flow-cytometric analysis showed that the percentage of TT population is S phase was 49% and 33% of the LoVo and CEM cell populations, respectively, while the G1/G0 fraction of TT was about 63% and 61% higher than that of LoVo and CEM, respectively. Our data suggest that the intrinsic chemoresistance in TT cells may be attributed to the combined factors of overexpression of the mdrl gene, a slower growth rate and a smaller proliferation fraction, although other factors or mechanisms that are yet to be investigated may also act in concert to contribute to the resistance phenotype of TT.

C-myc, N-myc, N-ras, and c-erb-B: lack of amplification or rearrangement in human medullary thyroid carcinoma and a derivative cell line.

We investigated the copy number and possible rearrangement of the four protooncogenes, c-myc, N-myc, N-ras, and c-erb-B, in DNA from seven untreated primary cancers or metastases of medullary thyroid carcinoma and an established human medullary thyroid carcinoma cell line, TT, using the Southern blotting technique. The purpose of this study was two-fold: 1) to examine whether protooncogene perturbations in medullary thyroid carcinoma could be considered as a prognostic marker; and 2) to determine whether the protooncogenes could have a possible role in medullary thyroid tumorigenesis. Neither amplification nor rearrangement of the protooncogenes was detectable in the DNA from any tumor samples or in the cell line. Our results suggest that DNA-evident amplification and rearrangement of the c-myc, N-myc, N-ras, and c-erb-B oncogenes may not be mechanisms through which these oncogenes become activated in this malignancy.

Deletion mapping on the distal third region of chromosome 1p in multiple endocrine neoplasia type IIA.

Multiple endocrine neoplasia type IIA (MEN IIA) syndrome is an autosomal-dominant endocrine disorder that consists of medullary thyroid carcinoma (MTC), pheochromocytoma, and parathyroid hyperplasia. The susceptibility gene to this disorder has been mapped to chromosome 10. However, molecular studies of tumor cells from patients with familial and sporadic MTC and/or pheochromocytoma have shown a high frequency (50%) of abnormalities on chromosome 1p. In the present study, we examined MTC or pheochromocytoma tumor specimens from eight patients (familial and nonfamilial cases) to investigate gene losses on chromosomes 1 and 10 as potential mechanisms for the tumors' development. The patients studies had homozygous genotypes in their leukocyte DNAs for the chromosome 10 marker used in this study, and the patients were, therefore, uninformative. However, the patients were informative for the chromosome 1 markers and five of the patients' tumor-cell DNAs (63%) had allelic deletions at one or multiple loci on chromosome 1p, and one tumor DNA had evidence of possible gene rearrangement; in all six cases, the abnormalities involved the distal third of chromosome 1p. Furthermore, we determined that the common break point for the 1p deletions was at 1p32. These results suggest that a tumor suppressor gene in this defined region is involved in the development/progression of MTC and pheochromocytoma by being either lost or inactivated.

Computerized classification of temporomandibular joint sounds.

Sounds, such as clicking and/or crepitation, evoked in the temporomandibular (jaw) joint during function may indicate pathology. Analysis of the reduced interference time-frequency distribution of these sounds is of diagnostic value. However, visual evaluation is expensive and error prone, and there is, thus, a need for automated analysis. The aim of this study was to find the optimal signal representation and pattern recognition method for computerized classification of temporomandibular joint sounds. Concepts of time-shift invariance with and without scale invariance were employed and mutually compared. The automated analysis methods provided classification results that were similar to previous visual classification of the sounds. It was found that the classifier performance was significantly improved when scale invariance was omitted. This behavior occurred because scale invariance interfered with the frequency content of the signal. Therefore, scale invariance should not be pursued in the classification scheme employed in this study.

Holistic outcome measurement for terminally ill cancer patients in medical centers in Taiwan.

The purpose of this cross-sectional study was to examine holistic patient outcomes for terminally ill cancer patients, as well as to examine whether different care patterns affect patient outcomes differently. Holistic patient outcomes were measured by the patients' quality of life, satisfaction with care, and cost of care. A purposive sampling of 224 subjects including 123 patients and 101 nurses was drawn from four medical centers in Taiwan. Among these settings, various care patterns were adopted and categorized into 4 groups: hospice inpatients, hospice team consultation, home hospice care services, and a conventional acute care group. Results showed that hospice inpatients had a higher quality of life, a higher level of satisfaction with the care and a lower average inpatient cost, whereas conventional care tended to have the highest length of hospital stay. Home hospice care patients had better psychological well-being than those with other care patterns. In addition, nurses' work satisfaction with the inpatients care unit tended to be significantly higher than with the other groups. The study findings not only provide an instrument for evaluating the quality of care, but also contribute to identifying patterns of care that will influence the dying process, which can only be beneficial for patients. Given the wide variety of healthcare services available now, understanding and selecting the most effective care patterns to enhance patient outcomes is of utmost importance in Taiwan.

[11 beta-Hydroxysteroid dehydrogenase].

There are two types of 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD). 11 beta-HSD 1 is an oxoreductase interconverting biologically active cortisol and inactive cortisone. 11 beta-HSD 2 is an exclusive oxidase only converting cortisol to cortisone. It has been recognized that 11 beta-HSD 2 confers the specificity of mineralocorticoid receptor in the kidney and protects the fetus from high levels of maternal glucocorticoids in the placenta. Lack or malfunction of this enzyme could result in the development of apparent mineralocorticoid excess syndrome and intrauterine growth retardation of the fetus. 11 beta-HSD is possibly involved in a number of other physiological and pathological conditions such as stress, hypertension, diabetes mellitus and neurodegenerative disorders. The functions of 11 beta-HSD 1 are not very well understood.