cyclic-gmp and Uterine-Cervical-Neoplasms

This paper investigates the function of lncRNA DARS-AS1 in cervical cancer (CC) as well as its in-depth mechanism. The differential expression of DARS-AS1 and ATP1B2 were analyzed based on The Cancer Genome Atlas and the Genotype-Tissue Expression databases, and the survival rate was measured using Kaplan-Meier survival analysis. Biological function experiments were performed to detect cell proliferation, invasion, and migration. Quantitative real-time polymerase chain reaction was carried out to detect the expression of DARS-AS1 and ATP1B2. Western blot analysis was utilized to assess the protein levels of ATP1B2 and cGMP-PKG pathway-related proteins. DARS-AS1 was expressed at high levels in CC tissues and cell lines, and high expression of DARS-AS1 indicated a lower survival rate. CCK-8 and colony formation assays revealed that the overexpression of DARS-AS1 promoted the proliferation of CC cells. Furthermore, bioinformatics analysis suggested that the cGMP-PKG pathway ranks as the first pathway enriched by the differential genes that correlated with DARS-AS1 (|r| > 0.4). ATP1B2, as a cGMP-PKG pathway-related gene, was significantly correlated with the overall survival of CC patients. We further confirmed that ATP1B2 was lowly expressed in CC and negatively correlated with the DARS-AS1 expression. Then, biological function experiments exhibited that the promotion of cell proliferation, invasion, and migration resulted due to the upregulation of DARS-AS1 could be canceled by ATP1B2 overexpression. Finally, Western blot revealed that upregulation of DARS-AS1 could activate the cGMP-PKG pathway, while overexpression of ATP1B2 reversed this activation. Our study revealed that DARS-AS1/ATP1B2 contributes to regulating the progression of CC at least partially by modulating the cGMP-PKG pathway.

Topics: Cyclic GMP; Cyclic GMP-Dependent Protein Kinases; Female; Humans; Neoplasm Proteins; RNA, Long Noncoding; RNA, Neoplasm; Second Messenger Systems; Uterine Cervical Neoplasms

To observe the effect of propranolol in cervical cancer and investigate the mechanism of the effect． METHODS AND RESULTS: We found 5 direct protein targets (DPTs) of propranolol (PRO) by DrugBank5.0 firstly. Next, we analyzed protein-protein interaction (PPI) network and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways of PRO DPTs and the result showed that PRO was linked with cGMP/PKG pathway. Then, we recognized the top 38 upexpressed genes of cervical cancer (CC) based original microarray datasets (GSE7803, GSE9750, GSE39001 and GSE63514). Further, we analyzed the biological process with the 38 overexpressed genes by STRING. We found some of overexpressed genes of CC participated in GMP biosynthetic process. Lastly, the function of PRO in CC was validated by MTT assay, Western blotting, flow cytometry and colony formation assay methods. We verified PRO can suppress cGMP/PKG pathway then inhibits CC cell growth.. The bioinformatical analysis combine with traditional experiment can help us understanding potential molecular mechanism about how PRO acting in CC. This method is a new paradigm which can guide future researches about mechanism in existing diseases and drugs.

Topics: Apoptosis; Cell Line, Tumor; Cell Proliferation; Cyclic GMP; Cyclic GMP-Dependent Protein Kinases; Female; HeLa Cells; Humans; Propranolol; Protein Interaction Maps; Signal Transduction; Uterine Cervical Neoplasms

We previously reported that human squamous cell carcinoma (SCC) cell lines refractory to cis-diaminedichloro-platinum II (cisplatin [CDDP]) had significant upregulation of the phosphodiesterase 3B gene (PDE3B), suggesting that inhibiting PDE3B suppresses CDDP resistance. shRNA-mediated PDE3B depletion in CDDP-resistant cells derived from SCC cells and Hela cells and induced CDDP sensitivity and inhibited tumor growth with elevated cyclic GMP induction resulting in upregulation of the multidrug-resistant molecule, but this did not occur in the 5-fluorouracil-resistant hepatocellular carcinoma cell lines. Furthermore, the antitumor growth effect of the combination of a PDE3B inhibitor (cilostazol) and CDDP in vivo was also greater than with either cilostazol or CDDP alone, with a significant increase in the number of apoptotic and cell growth-suppressive cancer cells in CDDP-resistance cell lines. Our results provided novel information on which to base further mechanistic studies of CDDP sensitization by inhibiting PDE3B in human cancer cells and for developing strategies to improve outcomes with concurrent chemotherapy.

Topics: Animals; Antineoplastic Combined Chemotherapy Protocols; Apoptosis; Body Weight; Carcinoma, Squamous Cell; Cilostazol; Cisplatin; Cyclic AMP; Cyclic GMP; Cyclic Nucleotide Phosphodiesterases, Type 3; Dose-Response Relationship, Drug; Drug Resistance, Neoplasm; Female; Gene Expression Regulation, Enzymologic; Gene Silencing; HeLa Cells; Humans; Mice; Mice, Nude; Phosphodiesterase 3 Inhibitors; RNA, Messenger; Tetrazoles; Tumor Cells, Cultured; Uterine Cervical Neoplasms; Xenograft Model Antitumor Assays

Carcinoma of the uterine cervix represents the second most frequent female malignancy worldwide, but few biochemical tumour markers have been implemented into clinical practice. Elevated extracellular guanosine 3', 5'-cyclic monophosphate (cGMP) levels have been reported to be a sensitive, early and reliable marker for screening relapse in carcinoma of the uterine cervix. The mechanism behind this observation remains unknown. The possibility exists that the cancer cells develop resistance to the antiproliferative effect of high intracellular cGMP levels. The enhanced cGMP expression may originate from either an increase in cellular export capacity by increased expression of member 5 in subfamily C of ATP-Binding-Cassette transporters (ABCC5), or increased substrate (cGMP) levels for this pump. The latter situation occurs with increased expression of inducible nitric oxide synthase (iNOS) and/or soluble guanylyl cyclase (sGC) and/or reduced expression of member 5 of the cyclic nucleotide phosphodiesterases (PDE5). Four transformed human cell lines derived from carcinomas of the uterine cervix (C-4 I, C-33 A, SiHa and ME-180 cells) and one non-transformed human cell line (WI-38) were included in the study in order to unveil which biokinetic components are involved. The expressions of iNOS, sGC, PDE5 and ABCC5 in the initial and final phase of the exponential growth curve were compared. Assuming that the WI-38 control cells mimic the situation in a normal tissue, iNOS remains un-expressed during proliferation, and the expression of sGC is low but shows a clear increase during exponential growth. PDE5 is highly expressed and increases (≈130%) during growth whereas ABCC5 exhibited low to moderate expression, with a moderate increase (≈40%) during growth. The malignant cells exhibited moderate ABCC5 expression with a distinct increase during exponential growth, whereas PDE5 expression remained virtually unchanged. Dysregulation of the cGMP biokinetics in growing malignant cells may account for the elevation of extracellular cGMP observed in patients with carcinoma of the uterine cervix.

Topics: Cell Division; Cyclic GMP; Cyclic Nucleotide Phosphodiesterases, Type 5; Female; Gene Expression; Gene Expression Regulation, Neoplastic; Guanylate Cyclase; Humans; Multidrug Resistance-Associated Proteins; Nitric Oxide Synthase Type II; Polymerase Chain Reaction; Receptors, Cytoplasmic and Nuclear; Soluble Guanylyl Cyclase; Uterine Cervical Neoplasms

Several studies have shown altered biokinetics of cyclic nucleotides in human cancer. In order to mimic the growing tumor bulk in carcinomas of the uterine cervix, four human cell lines (C4-I, C33A, ME-180, and SiHa) were expanded in serum-supplemented cell cultures. The extra- and intracellular levels of cAMP and cGMP were determined at increasing cell densities. In all of the cell lines, a cell density-dependent increase in the extracellular cGMP/cAMP ratio was observed. cAMP was distributed to the extracellular compartment against a concentration gradient at low cell densities but was retained in the intracellular compartment at high cell densities. In contrast, cGMP was distributed to the extracellular compartment against a concentration gradient for the whole range of cell densities. This study suggests that the cell density-dependent increase in the extracellular cGMP/cAMP ratio in cell lines derived from carcinomas of the uterine cervix is a result of changes occurring in both intracellular levels and cellular excretion of cyclic nucleotides.

Topics: Cell Count; Cell Division; Cyclic AMP; Cyclic GMP; Female; Humans; Tumor Cells, Cultured; Uterine Cervical Neoplasms

We show here that the human cervix carcinoma cell line ME-180 expresses a constitutive nitric oxide (NO) synthase, as demonstrated by formation of [3H]citrulline and nitrite. The enzyme is dependent on tetrahydrobiopterin, NADPH, flavins and Ca2+/calmodulin. Enzyme activity is located in the cytosol rather than in the membrane fraction and can be inhibited by NG-monomethyl-L-arginine (NMMA). An antiserum to NO synthase purified from porcine cerebellum inhibited the enzyme activity. ME-180 cells released NO, as was shown by stimulation of guanylate cyclase (EC 4.6.1.2) in RFL-6 detector cells; this release was stimulated 8-fold by the Ca2+ ionophore A23187 and 2-fold by increasing the intracellular tetrahydrobiopterin levels with cytokines. This is the first characterization of a Ca2+/calmodulin-dependent NO synthase activity in human epithelial-type tumour cells.

Topics: Amino Acid Oxidoreductases; Arginine; Calcium; Calmodulin; Cyclic GMP; Female; Humans; Interferon-gamma; Kinetics; Lipopolysaccharides; Nitric Oxide Synthase; omega-N-Methylarginine; Pteridines; Recombinant Proteins; Tumor Cells, Cultured; Uterine Cervical Neoplasms

The ratio between cGMP and cAMP in plasma/urine is elevated in several types of malignancies. The present in vitro study showed that the ratio between extracellular cGMP and cAMP increased during the proliferation of C4-I cells (derived from a carcinoma of the uterine cervix), whereas this ratio decreased in WI-38 cells (normal lung fibroblasts). These results can be explained by differences between the transformed and non-transformed cells in the cell-density-dependent transmembrane distribution and intracellular levels of cyclic nucleotides. In the serum-deprived cultures, no profound effects were seen on the cell-density-dependent biokinetics of cAMP and cGMP. In the absence of serum, growth of C4-I cells was markedly retarded, whereas WI-38 cells were unable to expand at all.

Topics: Cell Count; Cell Division; Cell Line, Transformed; Culture Media; Culture Media, Serum-Free; Cyclic AMP; Cyclic GMP; Extracellular Space; Female; Humans; Intracellular Fluid; Lung; Uterine Cervical Neoplasms

Low sensitizing doses given to patients with cervical carcinoma in the course of complex radiation treatment were responsible for a greater decrease in the frequency of radiation injuries to the intestine as compared with standard complex radiation procedure. This was matched by an increase in the incidence of exacerbation of chronic adnexitis. Radiation appeared to inhibit thyroid function. Low sensitizing dose treatment given in the course of complex radiotherapy did not affect the gland's function. However, cases of stage III cancer exhibited a more pronounced decrease in thyrotrophin levels accompanied by higher somatotropin levels.

Topics: Brachytherapy; Combined Modality Therapy; Cyclic AMP; Cyclic GMP; Female; Homeostasis; Hormones; Humans; Uterine Cervical Neoplasms

Blinded urinary assays for cyclic guanosine 3':5'-monophosphate (cGMP) and cyclic adenosine 3':5'-monophosphate (cAMP) were performed on 49 subjects with documented abnormal cervical cytology and 21 control subjects with normal cytology. A significant difference in the mean cGMP:cAMP ratios between the case and control groups was found. A significantly greater proportion of women with cytological abnormalities had a cGMP:cAMP ratio above the 0.2 level (p less than 0.001). Cases treated surgically for severe dysplasia or carcinoma in situ of the cervix revealed a significant postsurgical fall in the cGMP:cAMP ratios (p less than 0.025). The possibility of utilizing urinary ratios of cyclic nucleotides as an objective index in the detection, monitoring of progression, and therapy of preneoplastic cervical lesions is discussed.

Topics: Carcinoma in Situ; Cyclic AMP; Cyclic GMP; Female; Humans; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms; Uterine Cervicitis

Cyclic guanosine 3',5' monophosphate (cyclic GMP) and cyclic adenosine 3',5' monophosphate (cyclic AMP) have been determined in random urine specimens from 95 healthy individuals, 60 patients with non-cancerous conditions, 52 patients with benign tumours, and 74 patients with malignant tumours. Concentrations of cyclic GMP have also been determined in a number of other groups, including some undergoing cancer treatment. Ninety-three per cent of cancer patients had raised urinary cyclic GMP concentrations compared to the reference range for healthy subjects. For the non-cancerous and benign groups, 33% and 42% respectively had raised concentrations. The urine cyclic AMP concentrations were similar in all groups. Urine cyclic GMP appeared to rise early in the onset of malignant growth. Successful cancer treatment was accompanied by a dramatic fall in the urine cyclic GMP concentrations, whereas if the treatment was unsuccessful the level did not change. It is concluded that urine cyclic GMP may have important applications in the monitoring of cancer treatment.

Topics: Adolescent; Adult; Aged; Breast Neoplasms; Cyclic AMP; Cyclic GMP; Female; Humans; Male; Middle Aged; Neoplasms; Ovarian Neoplasms; Uterine Cervical Neoplasms; Uterine Prolapse