Cervical Carcinoma Cell Line HeLa Research Articles

Characterization and Prediction of Lysine (K)-Acetyl-Transferase Specific Acetylation Sites

Lysine acetylation is a well-studied post-translational modification on both histone and nonhistone proteins. More than 2000 acetylated proteins and 4000 lysine acetylation sites have been identified by large scale mass spectrometry or traditional experimental methods. Although over 20 lysine (K)-acetyl-transferases (KATs) have been characterized, which KAT is responsible for a given protein or lysine site acetylation is mostly unknown. In this work, we collected KAT-specific acetylation sites manually and analyzed sequence features surrounding the acetylated lysine of substrates from three main KAT families (CBP/p300, GCN5/PCAF, and the MYST family). We found that each of the three KAT families acetylates lysines with different sequence features. Based on these differences, we developed a computer program, Acetylation Set Enrichment Based method to predict which KAT-families are responsible for acetylation of a given protein or lysine site. Finally, we evaluated the efficiency of our method, and experimentally detected four proteins that were predicted to be acetylated by two KAT families when one representative member of the KAT family is over expressed. We conclude that our approach, combined with more traditional experimental methods, may be useful for identifying KAT families responsible for acetylated substrates proteome-wide.

The Human Papillomavirus E6 Oncogene Represses a Cell Adhesion Pathway and Disrupts Focal Adhesion through Degradation of TAp63β upon Transformation

Cervical carcinomas result from cellular transformation by the human papillomavirus (HPV) E6 and E7 oncogenes which are constitutively expressed in cancer cells. The E6 oncogene degrades p53 thereby modulating a large set of p53 target genes as shown previously in the cervical carcinoma cell line HeLa. Here we show that the TAp63β isoform of the p63 transcription factor is also a target of E6. The p63 gene plays an essential role in skin homeostasis and is expressed as at least six isoforms. One of these isoforms, ΔNp63α, has been found overexpressed in squamous cell carcinomas and is shown here to be constitutively expressed in Caski cells associated with HPV16. We therefore explored the role of p63 in these cells by performing microarray analyses after repression of endogenous E6/E7 expression. Upon repression of the oncogenes, a large set of p53 target genes was found activated together with many p63 target genes related to cell adhesion. However, through siRNA silencing and ectopic expression of various p63 isoforms we demonstrated that TAp63β is involved in activation of this cell adhesion pathway instead of the constitutively expressed ΔNp63α and β. Furthermore, we showed in cotransfection experiments, combined with E6AP siRNA silencing, that E6 induces an accelerated degradation of TAp63β although not through the E6AP ubiquitin ligase used for degradation of p53. Repression of E6 transcription also induces stabilization of endogenous TAp63β in cervical carcinoma cells that lead to an increased concentration of focal adhesions at the cell surface. Consequently, TAp63β is the only p63 isoform suppressed by E6 in cervical carcinoma as demonstrated previously for p53. Down-modulation of focal adhesions through disruption of TAp63β therefore appears as a novel E6-dependent pathway in transformation. These findings identify a major physiological role for TAp63β in anchorage independent growth that might represent a new critical pathway in human carcinogenesis.

Synthesis of polyfunctionalized piperidone oxime ethers and their cytotoxicity on HeLa cells

A series of twenty 2,6-diarylpiperidin-4-one O-methyloximes were synthesized with fluoro/chloro/bromo/methyl/methoxy/ethoxy/isopropyl substituents on various positions of the phenyl at C-2 and C-6 in association with/without methyl substituent on the secondary amino group and methyl/ethyl/isopropyl substituents on the active methylene centers. Regardless of their substitution all compounds predominantly exist in the chair conformation except 3m, which adopts a twist-boat conformation. All the synthesized compounds were evaluated for their in vitro antiproliferative activity against human cervical carcinoma (HeLa) cell line. The cytotoxicity of the test compounds was determined by measuring the number of live cells after 24 h of treatment by MTT assay method. This preliminary SAR suggests some lead molecules 3c-f, 3j-k, 4d-g, and 4i with a scope of further structural optimization of the piperidone pharmacophore toward the development of anticancer drug synthesis.

Effects of Buthus martensii Karsch Venom on Cell Proliferation and Cytotoxicity in Hela Cells

This paper focuses on the effect of the venom of the scorpion Buthus martensii on the proliferation of human cervical carcinoma Hela cell line and the related molecular mechanism. MTT test showed that the scorpion venom inhibited proliferation of Hela cells in time-dependent and concentration-dependent manner with 50% inhibitory concentration (IC50) of 34.5 μg/mL(48 h). By using flow cytometry, it was found that the scorpion venom could induce apoptosis and necrosis in Hela cells. RT-PCR and Western blot indicated there were obviously up-regulated in the expressions of p21 protein but the expression of p21 mRNA showed no significant difference in the Hela cell by the scorpion venom. These results suggest that the possible mechanism of the scorpion venom is to activate the expressions of p21 protein and to cause Hela cell apoptosis.

CCAAT/Enhancer-binding Protein δ Mediates Tumor Necrosis Factor α-induced Aurora Kinase C Transcription and Promotes Genomic Instability

Epidemiologic and clinical research indicates that chronic inflammation increases the risk of certain cancers, possibly through chromosomal instability. However, the mechanism of inflammation-dependent chromosomal instability associated with tumorigenesis is not well characterized. The transcription factor CCAAT/enhancer-binding protein δ (C/EBPδ, CEBPD) is induced by tumor necrosis factor α (TNFα) and expressed in chronically inflamed tissue. In this study, we show that TNFα promotes aneuploidy. Loss of CEBPD attenuated TNFα-induced aneuploidy, and CEBPD caused centromere abnormality. Additionally, TNFα-induced CEBPD expression augmented anchorage-independent growth. We found that TNFα induced expression of aurora kinase C (AURKC) through CEBPD, and that AURKC also causes aneuploidy. Furthermore, high CEBPD expression correlated with AURKC expression in inflamed cervical tissue specimens. These data provide insight into a novel function for CEBPD in inducing genomic instability through the activation of AURKC expression in response to inflammatory signals.

Suppression of HPV E6 and E7 expression by BAF53 depletion in cervical cancer cells

Deregulation of the expression of human papillomavirus (HPV) oncogenes E6 and E7 plays a pivotal role in cervical carcinogenesis because the E6 and E7 proteins neutralize p53 and Rb tumor suppressor pathways, respectively. In approximately 90% of all cervical carcinomas, HPVs are found to be integrated into the host genome. Following integration, the core-enhancer element and P105 promoter that control expression of E6 and E7 adopt a chromatin structure that is different from that of episomal HPV, and this has been proposed to contribute to activation of E6 and E7 expression. However, the molecular basis underlying this chromatin structural change remains unknown. Previously, BAF53 has been shown to be essential for the integrity of higher-order chromatin structure and interchromosomal interactions. Here, we examined whether BAF53 is required for activated expression of E6 and E7 genes. We found that BAF53 knockdown led to suppression of expression of E6 and E7 genes from HPV integrants in cervical carcinoma cell lines HeLa and SiHa. Conversely, expression of transiently transfected HPV18-LCR-Luciferase was not suppressed by BAF53 knockdown. The level of the active histone marks H3K9Ac and H4K12Ac on the P105 promoter of integrated HPV 18 was decreased in BAF53 knockdown cells. BAF53 knockdown restored the p53-dependent signaling pathway in HeLa and SiHa cells. These results suggest that activated expression of the E6 and E7 genes of integrated HPV is dependent on BAF53-dependent higher-order chromatin structure or nuclear motor activity.

RNA interference targeting protein kinase C iota inhibits cell proliferation of cervical carcinoma cell line HeLa

Objective To explore the effect of RNA interference （RNAi） targeting protein kinase C iota （PKCl） on proliferation and apoptosis of cervical carcinoma cell line HeLa. Methods Stable cell lines which were transfected with PKCl shRNA or vector-mock plasmids were established. The cell lines, psi-PKCiota-HeLa, psi-mock-HeLa and HeLa, were used to detect methyl thiazol tetrazolium （MTT） absorbance at 1 st-5th day. The cells were cultured in 6-well plates and stained by crystal violent at 21 st day, and the stained cells colonies were counted. Flow cytometry was used to detect apoptosis and cell cycle distribution after HeLa cells were transiently transfected with PKC iota shRNA at 72nd h. Results Compared with the psi-mock and blank controls, the transfected cells stably expressing PKCl shRNA showed markedly decrease of proliferation assayed by MTT and plate colony formation. The apoptosis rate of cells transiently transfected with PKCL shRNA （9. 54 ± 0. 55 ） % was higher than that of cells transiently transfected with psi-mock （ 1.31 ±0. 10） % or HeLa blank control calls （2. 75 ±0. 24） % （ P 〈 0. 05 ）. The cells transiently transfected by PKCl shRNA were arrested in G0/G1 phase. Conclusion PKCl is essential for maintaining of HeLa cell proliferation. Silencing PKCl can inhibit the growth of HeLa cells and induce apoptosis. Key words: Cervical carcinoma ; RNA interference ; Proliferation ; Apoptosis

ChemInform Abstract: Two Novel Benzastatin Derivatives, JBIR‐67 and JBIR‐73, Isolated from Streptomyces sp. RI18.

AbstractThe new benzastatin derivatives, JBIR‐67 (I) and JBIR‐73 (II) show weak cytotoxicity against human cervical carcinoma HeLa cell lines.

Honeycomb-Structured Films by Multifunctional Amphiphilic Biodegradable Copolymers: Surface Morphology Control and Biomedical Application as Scaffolds for Cell Growth

Recently, fabrication of functional porous polymer films with patterned surface structures at the scale from nanometer to micrometer has been attracting increasing interests in material science and nanobiotechnology. In this work, we present new preparation of two series of multifunctional amphiphilic copolymers and preparation of their microporous thin films on solid substrates. First, diblock dendritic poly(l-lysine)-b-poly(l-lactide)s and triblock dendritic poly(l-lysine)-b-poly(l-lactide)-b-dendritic poly(l-lysine)s (C1-C6) were synthesized through 4-dimethylaminopyridine (DMAP)-catalyzed living ring-opening polymerization of (l-)-lactide with (l-)-lysine dendron initiators, and their structures were characterized by nuclear magnetic resonance spectrometer (NMR), gel permeation chromatography (GPC) and matrix-assisted laser desorption/ionization Fourier-transformed mass spectra (MALDI-FTMS). Employing the breath-figure (BF) fabrication strategy, thin films of the synthesized amphiphiles (C1-C6) were drop-cast, and their surface topologies were examined by scanning electron microscopy (SEM) and atomic force microscopy (AFM), and the effects of new amphiphile structure and drop-casting parameters of amphiphile concentration, humidity and temperature on self-assembly of ordered porous surface were studied. Furthermore, the influence of surface energy of drop-casting substrates was additionally investigated. With a human cervical epithelial carcinoma cell line (HeLa), cytotoxicity of the prepared honeycomb-structured films by new amphiphile C6 was evaluated by thiazoyl-blue-tetrazolium-bromide (MTT) assay, and HeLa cell growth behavior with microporous amphiphile films as the scaffolds was also examined. It was found that tunable micropore diameter sizes and well ordered surface topologies of BF films could be achieved for the new prepared amphiphiles, and utilization of the honeycomb-like microporous films as scaffolds indicated favorable enhancement in cell proliferation. Therefore, the honeycomb-structured films by these biocompatible multifunctional amphiphiles may provide new materials as 3D-scaffold materials for potential application in tissue engineering and regeneration.

Antioxidant activity and cytotoxic effect of aviprin and aviprin-3″-O-D-glucopyranoside on LNCaP and HeLa cell lines

Many researchers have shown that plant-derived polyphenolic compounds are helpful nutraceuticals in restraining various disorders such as neoplastic diseases. In this study two linear furanocoumarins, aviprin and aviprin-3″-O-D-glucopyranoside (A3G), were isolated from methanol extract of Prangos uloptera roots. The evaluation of free radical scavenging capacity of the compounds showed that aviprin is a more effective antioxidant than A3G with RC50 of 0.54 mg mL−1. The biological and antiproliferative activities of the furanocoumarins were examined using human cervical carcinoma HeLa cell line and LNCaP prostatic cell line. Cell membrane integrity and cell viability were evaluated by measuring trypan blue exclusion assay and reduction of the tetrazolium blue compound, respectively. Treating the LNCaP cell line with various concentrations of the furanocoumarins showed that IC50 of aviprin and A3G were 0.4 and 6.6 mg mL−1, whereas their CC50 values were 0.7 and 11 mg mL−1, respectively. These results indicated that 42.7% of LNCaP cells were not dead by necrosis. Treating the HeLa cells by the furanocoumarins showed the greater sensitivity of the HeLa cell line than the LNCaP cell line. A morphological analysis and the study of DNA fragmentation provided further some evidence for the inhibition of the LNCaP cell line via apoptosis induction.

TRAIL-R4 Promotes Tumor Growth and Resistance to Apoptosis in Cervical Carcinoma HeLa Cells through AKT

BackgroundTRAIL/Apo2L is a pro-apoptotic ligand of the TNF family that engages the apoptotic machinery through two pro-apoptotic receptors, TRAIL-R1 and TRAIL-R2. This cell death program is tightly controlled by two antagonistic receptors, TRAIL-R3 and TRAIL-R4, both devoid of a functional death domain, an intracellular region of the receptor, required for the recruitment and the activation of initiator caspases. Upon TRAIL-binding, TRAIL-R4 forms a heteromeric complex with the agonistic receptor TRAIL-R2 leading to reduced caspase-8 activation and apoptosis.Methodology/Principal FindingsWe provide evidence that TRAIL-R4 can also exhibit, in a ligand independent manner, signaling properties in the cervical carcinoma cell line HeLa, through Akt. Ectopic expression of TRAIL-R4 in HeLa cells induced morphological changes, with cell rounding, loss of adherence and markedly enhanced cell proliferation in vitro and tumor growth in vivo. Disruption of the PI3K/Akt pathway using the pharmacological inhibitor LY294002, siRNA targeting the p85 regulatory subunit of phosphatidylinositol-3 kinase, or by PTEN over-expression, partially restored TRAIL-mediated apoptosis in these cells. Moreover, the Akt inhibitor, LY294002, restituted normal cell proliferation index in HeLa cells expressing TRAIL-R4.Conclusions/SignificanceAltogether, these results indicate that, besides its ability to directly inhibit TRAIL-induced cell death at the membrane, TRAIL-R4 can also trigger the activation of signaling pathways leading to cell survival and proliferation in HeLa cells. Our findings raise the possibility that TRAIL-R4 may contribute to cervical carcinogenesis.

Synthesis, characterization, and in vitro biological evaluation of highly stable diversely functionalized superparamagnetic iron oxide nanoparticles

In this article, we report the design and synthesis of a series of well-dispersed superparamagnetic iron oxide nanoparticles (SPIONs) using chitosan as a surface modifying agent to develop a potential T 2 contrast probe for magnetic resonance imaging (MRI). The amine, carboxyl, hydroxyl, and thiol functionalities were introduced on chitosan-coated magnetic probe via simple reactions with small reactive organic molecules to afford a series of biofunctionalized nanoparticles. Physico-chemical characterizations of these functionalized nanoparticles were performed by TEM, XRD, DLS, FTIR, and VSM. The colloidal stability of these functionalized iron oxide nanoparticles was investigated in presence of phosphate buffer saline, high salt concentrations and different cell media for 1 week. MRI analysis of human cervical carcinoma (HeLa) cell lines treated with nanoparticles elucidated that the amine-functionalized nanoparticles exhibited higher amount of signal darkening and lower T 2 relaxation in comparison to the others. The cellular internalization efficacy of these functionalized SPIONs was also investigated with HeLa cancer cell line by magnetically activated cell sorting (MACS) and fluorescence microscopy and results established selectively higher internalization efficacy of amine-functionalized nanoparticles to cancer cells. These positive attributes demonstrated that these nanoconjugates can be used as a promising platform for further in vitro and in vivo biological evaluations.

The new 5- or 6-azapyrimidine and cyanuric acid derivatives of l-ascorbic acid bearing the free C-5 hydroxy or C-4 amino group at the ethylenic spacer: CD-spectral absolute configuration determination and biological activity evaluations

We report on the synthesis of the novel types of cytosine and 5-azacytosine ( 1– 9), uracil and 6-azauracil ( 13– 18) and cyanuric acid ( 19– 22) derivatives of l-ascorbic acid, and on their cytostatic activity evaluation in human malignant tumour cell lines vs. their cytotoxic effects on human normal fibroblasts (WI38). The CD spectra analysis revealed that cytosine ( 5 and 6), uracil ( 14– 16), 6-azauracil ( 17) and cyanuric acid ( 21) derivatives of l-ascorbic acid bearing free amino group at ethylenic spacer existed as a racemic mixture of enantiomers, whereas L-ascorbic derivatives containing the C-5 substituted hydroxy group at the ethylenic spacer were obtained in (4 R, 5 S) enantiomeric form. The stereochemistry of 6-azauracil derivative of l-ascorbic acid ( 13) was confirmed by X-ray crystal structure analysis. The molecules are self-assembled by one N–H⋯O hydrogen bond, two C–H⋯O hydrogen bonds and two C–H⋯π interactions into three-dimensional framework. Cytostatic activity evaluation indicated that compounds did not show distinctive antiproliferative effects on tested cell line panel. However, the cytosine derivative of l-ascorbic acid ( 1) containing the C4- C5 double bond conjugated with the lactone moiety produced rather marked growth inhibitory effect on hepatocellular carcinoma (HepG2), metastatic breast epithelial carcinoma (MCF-7) and cervical carcinoma (HeLa) cell lines at micromolar concentrations, but also exerted strong cytostatic effect on WI38. 5-Azacytosine derivative of l-ascorbic acid ( 2) with a double bond at the C4– C5 conjugated with the lactone moiety displayed potent antitumour activity against tested tumour cell lines with meanIC 50 values ranging from 0.92 to 5.91 μM. However, this compound also exhibited pronounced cytotoxicity towards WI38. Flow cytometric analysis of the cell cycle revealed that compound 2 triggers S phase arrest, which clearly demonstrates its interference with DNA replication, a key event of cell proliferation. Marked anticancer efficacy of compound 2 supports further in vivo investigation into its possible clinical utility.

Veraguamides A−G, Cyclic Hexadepsipeptides from a Dolastatin 16-Producing Cyanobacterium Symploca cf. hydnoides from Guam

Cytotoxicity-directed purification of a Symploca cf. hydnoides sample from Cetti Bay, Guam, afforded seven new cyclic depsipeptides, veraguamides A-G (1-7), together with the known compound dolastatin 16. The planar structures of 1-7 were elucidated using NMR and MS experiments, while enantioselective HPLC and Mosher's analysis of acid and base hydrolysates, respectively, were utilized to assign the absolute configurations of the stereocenters. Veraguamides A-G (1-7) are characterized by the presence of an invariant proline residue, multiple N-methylated amino acids, an α-hydroxy acid, and a C8-polyketide-derived β-hydroxy acid moiety with a characteristic terminus as either an alkynyl bromide, alkyne, or vinyl group. These compounds and a semisynthetic analogue (8) showed moderate to weak cytotoxic activity against HT29 colorectal adenocarcinoma and HeLa cervical carcinoma cell lines. Preliminary structure-activity relationship analysis identified several sensitive positions in the veraguamide scaffold that affect the cytotoxic activity of this compound class. Dolastatin 16 showed only weak cytotoxic activity on both cell lines tested. The complete stereostructure of dolastatin 16 was proposed for the first time through degradation followed by a combination of advanced Marfey's analysis and modified Mosher's analysis using phenylglycine methyl ester as a chiral anisotropic reagent.

Liposome Encapsulation of Thiol-Capped CdTe Quantum Dots for Enhancing the Intracellular Delivery

Although water soluble thiol-capped quantum dots (QDs) have been widely used as photoluminescence (PL) probes in various applications, the negative charges on thiol terminals limit the cell uptake hindering their applications in cell imaging. The commercial liposome complex (Sofast®) was used to encapsulate these QDs forming the liposome vesicles with the loading efficiency as high as about 95%. The cell uptakes of unencapsulated QDs and QD loaded liposome vesicles were comparatively studied by a laser scanning confocal microscope. We found that QD loaded liposome vesicles can effectively enhance the intracellular delivery of QDs in three cell lines (human osteosarcoma cell line (U2OS); human cervical carcinoma cell line (Hela); human embryonic kidney cell line (293 T)). The photobleaching of encapsulated QDs in cells was also reduced comparing with that of unencapsulated QDs, measured by the PL decay of cellular QDs with a continuous laser irradiation in the microscope. The flow cytometric measurements further showed that the enhancing ratios of encapsulated QDs on cell uptake are about 4-8 times in 293 T and Hela cells. These results suggest that the cationic liposome encapsulation is an effective modality to enhance the intracellular delivery of thiol-capped QDs.

Anti-tumour activities and a high-performance liquid chromatography mass spectrometric method for analysis of the constituents of Lomatogonium carinthiacum

In the present study, extracts of CHCl3, n-BuOH and water of Lomatogonium carinthiacum were tested for their possible anticancer effects on human lung adenocarcinoma A549 cell line, human erythroleukaemia K562 cell line and human cervical carcinoma HeLa cell line. The inhibitory effect of the extracts on cell proliferation was assessed by MTT colourimetric assay in vitro. A high-performance liquid chromatography-electrospray-mass spectrometric (HPLC-EIS-MS/MS) method was developed for the determination of the constituents of the extracts. According to HPLC-EIS-MS/MS data, the chemical structures of 21 constituents of L. carinthiacum were identified on-line without time-consuming isolation. The L. carinthiacum extracts showed inhibitory effects on the abovementioned cell lines. Extracts of CHCl3 were found to be the most inhibitory, with IC50 values of 0.13, 0.75 and 0.60 µg mL−1 on A549, K562 and HeLa, respectively. According to the IC50 values, the order of sensitivity of the cell lines was A549 > HeLa > K562 and the inhibitory effects to the cell lines of these extracts were in the order CHCl3 extract > water extract > n-BuOH, as xanthones > iridoids and secoiridoids > flavonols. The present study showed inhibitory activity of L. carinthiacum extracts on tumour cells.

Regulation of Na +-coupled glucose carrier SGLT1 by human papillomavirus 18 E6 protein

Tumor cells utilize preferably glucose for energy production. They accomplish cellular glucose uptake in part through Na +-coupled glucose transport mediated by SGLT1 (SLC5A1). This study explored the possibility that the human papillomavirus 18 E6 protein HPV18 E6 (E6) participates in the stimulation of SGLT1 activity. E6 is one of the two major oncoproteins of high-risk human papillomaviruses, which are the causative agent for cervical carcinoma. According to Western blotting, SGLT1 is expressed in the HPV18-positive cervical carcinoma cell line HeLa. To explore whether E6 affects SGLT1 activity, SGLT1 was expressed in Xenopus oocytes with and without E6 and electrogenic glucose transport determined by dual electrode voltage clamp. In SGLT1-expressing oocytes, but not in oocytes injected with water or expressing E6 alone, glucose triggered a current ( I g). I g was significantly increased by coexpression of E6 but not by coexpression of E2. According to chemiluminescence and confocal microscopy, coexpression of E6 significantly increased the SGLT1 protein abundance in the cell membrane. The decay of I g following inhibition of carrier insertion by Brefeldine A (5 μM) was not significantly affected E6 coexpression. Accrodingly, E6 was not effective by increasing carrier protein stability in the membrane. In conclusion, HPV18 E6 oncoprotein participates in the upregulation of SGLT1.

CHIP Promotes Human Telomerase Reverse Transcriptase Degradation and Negatively Regulates Telomerase Activity

The maintenance of eukaryotic telomeres requires telomerase, which is minimally composed of a telomerase reverse transcriptase (TERT) and an associated RNA component. Telomerase activity is tightly regulated by expression of human (h) TERT at both the transcriptional and post-translational levels. The Hsp90 and p23 molecular chaperones have been shown to associate with hTERT for the assembly of active telomerase. Here, we show that CHIP (C terminus of Hsc70-interacting protein) physically associates with hTERT in the cytoplasm and regulates the cellular abundance of hTERT through a ubiquitin-mediated degradation. Overexpression of CHIP prevents nuclear translocation of hTERT and promotes hTERT degradation in the cytoplasm, thereby inhibiting telomerase activity. In contrast, knockdown of endogenous CHIP results in the stabilization of cytoplasmic hTERT. However, it does not affect the level of nuclear hTERT and has no effect on telomerase activity and telomere length. We further show that the binding of CHIP and Hsp70 to hTERT inhibits nuclear translocation of hTERT by dissociating p23. However, Hsp90 binding to hTERT was not affected by CHIP overexpression. These results suggest that CHIP can remodel the hTERT-chaperone complexes. Finally, the amount of hTERT associated with CHIP peaks in G(2)/M phases but decreases during S phase, suggesting a cell cycle-dependent regulation of hTERT. Our data suggest that CHIP represents a new pathway for modulating telomerase activity in cancer.

Inhibin-betaA and -betaB subunits in normal and malignant glandular epithelium of uterine cervix and HeLa cervical cancer cell line

Inhibins, dimeric peptide hormones composed of an alpha-subunit and one of two possible beta-subunits (betaA or betaB), exhibit substantial roles in human reproduction and in endocrine-responsive tumors. However, it is still unclear if normal and cancerous cervical glandular epithelial cells as well as cervical cancer cell lines of glandular origin express the inhibin-betaA and -betaB subunits. Normal cervical tissue samples and a total of 10 specimens of well-differentiated adenocarcinomas of the human cervix were analyzed for inhibin-betaA and -betaB subunit expression by immunohistochemical analysis. Additionally, the cervical carcinoma cell line HeLa was analyzed by immunofluorescence and RT-PCR analysis for the expression of inhibin subunits. Immunolabeling of normal and malignant glandular epithelium of human cervical tissue revealed a positive staining reaction for the inhibin-betaA and -betaB subunits. Additionally, the cancer cell line HeLa synthesized both inhibin subunits. When compared to the normal cervical glandular epithelium, the expression of the inhibin beta subunits became significantly reduced in cervical adenocarcinoma tissues. In conclusion, we demonstrated a strong, though differential expression pattern of inhibin-betaA and -betaB subunits in normal and malignant glandular epithelial cells of the human uterine cervix. Although the physiological role of inhibins is still quite unclear in cervical tissue, the expression of inhibin-beta-subunits might play an important role in cervical cancer carcinogenesis, since they are significantly down-regulated during pathogenesis in cervical adenocarcinomas.

Effect of ferric ions on reactive oxygen species formation, cervical cancer cell lines growth and E6/E7 oncogene expression

As iron ions may participate in the pathogenesis of cancer and viral infections, the aim of this study was to monitor their influence on proliferation, E6 and E7 oncogene expression and reactive oxygen species (ROS) production in two human papilloma virus (HPV) positive cervical carcinoma cell lines (HeLa and SiHa) and one HPV negative vulvar cell line (A431). The anti-anaemic drug, ferric–sorbitol–citric acid complex (FSC) as a source of Fe III ions was used. Cells were treated with FSC at the concentrations between 0.001 and 1 mM Fe III for different time periods. Fe III ions inhibited the viability of HeLa and A431 cells while it had no influence on SiHa cells. Furthermore, Fe III treatment showed a time-dependent and a higher stimulatory effect on E6/E7 expression in SiHa cells than in HeLa cells. Fe III ion treatment with concentrations lower than 0.1 mM showed a time and a concentration dependent intracellular ROS production in all tested cell lines, while the treatment with 1 mM concentration decreased ROS production in all tested cell lines. In conclusion, Fe III ion treatment apart from having an anti-tumour effect, as we previously described, enhances survival of HPV 16-positive cells and might be associated with HPV oncogenesis.