Cervical Carcinoma Cell Line HeLa Research Articles

Synthesis, Characterization, and in Vitro Evaluation of Long-Chain Hyperbranched Poly(ethylene glycol) as Drug Carrier

A series of novel long-chain hyperbranched poly(ethylene glycol)s (LHPEGs) with biodegradable connections were designed and synthesized in one pot through proton-transfer polymerization using PEG and commercial glycidyl methacrylate as monomers and potassium hydride as catalyst. The LHPEGs were hydrolyzed at neutral pH resulting in the decrease of molecular weights. In vitro evaluation demonstrated that LHPEGs were biocompatible and displayed negligible hemolytic activity. The efficient cellular uptake of LHPEGs was confirmed by flow cytometry and confocal laser scanning microscopy. Moreover, conjugation of a model hydrophobic anticancer drug methotrexate to LHPEGs inhibited the proliferation of a human cervical carcinoma Hela cell line. MTT assay indicated that the conjugated methotrexate dose required for 50% cellular growth inhibition against Hela cells was 20 μg/mL. By combining the advantages of long-chain hyperbranched structure and PEG, LHPEG provides a promising drug carrier for therapeutic fields.

Epidermal growth factor receptor expression affects the efficacy of the combined application of saponin and a targeted toxin on human cervical carcinoma cells

Cervical cancer is the second most common cancer in women worldwide. Targeting the epidermal growth factor receptor (EGFR) is a very promising approach since it is overexpressed in about 90% of cervical tumors. Here, we quantified the toxic effect of SE, a targeted toxin consisting of epidermal growth factor (EGF) as targeting moiety and the plant toxin saporin-3, on 3 common human cervical carcinoma cell lines (HeLa, CaSki and SiHa) and recently established lines (PHCC1 and PHCC2) from 2 different individuals. A human melanocytic and a mouse cell line served as negative control. Additionally, we combined SE with saponinum album, a saponin composite from Gypsophila paniculata, which exhibited synergistic properties in previous studies. The cell lines, except for SiHa cells, revealed high sensitivity to SE with 50% cell survival in the range of 5-24.5 nM. The combination with saponin resulted in a remarkable enhancement of cytotoxicity with enhancement factors ranging from 9,000-fold to 2,500,000-fold. The cytotoxicity of SE was clearly target receptor specific since free EGF blocks the effect and saporin-3 alone was considerably less toxic. For all cervical carcinoma cell lines, we evinced a clear correlation between EGFR expression and SE sensitivity. Our data indicate a potential use of targeted toxins for the treatment of cervical cancer. In particular, the combination with saponins is a promising approach since efficacy is drastically improved.

Antioxidant and cytotoxic effect of oxidized lutein in human cervical carcinoma cells (HeLa)

This study investigated the antioxidant and cytotoxic effect of oxidized lutein using human cervical carcinoma cell lines (HeLa). Liposome contained phosphatidylcholine (20μmol) in Tris–HCl buffer and lutein (200μmol) was exposed to sunlight for 100min. Photo-oxidized lutein products were characterized by LC–MS (APCI+) and studied for their antioxidant property and apoptosis in terms of cell viability, glutathione and malondialdehyde (MDA) levels. Photo-oxidized lutein fragmented ions were identified as 523 (M++H+-3CH3), 476 (M++H+-6CH3), 551 (M++H+-H2O) and its isomers as 13-Z lutein, 13′-Z lutein, 13-Z zeaxanthin, all-E zeaxanthin, 9-Z lutein, 9′-Z lutein. Free radical scavenging activity of oxidized lutein was higher by 45.9% (IC50, 3.71μg) than lutein (IC50, 5.28μg). Oxidized lutein lowered the lipid peroxidation by 20.7% than lutein. The viability of HeLa cells, glutathione and MDA levels were decreased by 64%, 40% and 18% than lutein. To conclude, oxidized lutein may be highly reactive, since oxidation results in radical ions, which can combine with similar reactive oxidative species that could lead to higher antioxidant effect. This may be true in this study that antioxidant property of oxidized lutein was higher than lutein that correlates with free radical scavenging activity and cytotoxic effects on HeLa cells.

Effect of siRNA‐mediated silencing of Bmi‐1 gene expression on HeLa cells

B-cell-specific Moloney murine leukemia virus insertion site 1 (Bmi-1) is highly expressed in several malignant tumors and its expression level is positively correlated with tumor invasion, distant metastasis, prognosis, and recurrence. In the present study, the biological effect of small interfering RNA (siRNA) that targeted Bmi-1 expression was studied in human cervical carcinoma cell line HeLa cells. Bmi-1 siRNA inhibited the expression of Bmi-1 at the mRNA and protein levels in HeLa cells, which significantly affected proliferation, colony formation, and migration of HeLa cells in vitro and in vivo. Therefore, silencing Bmi-1 may be a potential therapeutic option for the management of some human cancers.

Enhanced photodynamic effect of cobalt(iii) dipyridophenazine complex on thyrotropin receptor expressing HEK293 cells

Ternary cobalt(III) complexes [CoL(B)] (1-3) of a trianionic tetradentate phenolate-based ligand (L) and phenanthroline bases (B), viz. 1,10-phenanthroline (phen in 1), dipyridoquinoxaline (dpq in 2) and dipyridophenazine (dppz in 3) are synthesized, characterized from X-ray crystallographic, analytical and spectral techniques, and their utility in photodynamic therapy (PDT) of thyroid diseases caused by TSH receptor dysfunction is probed. The complexes display a visible spectral band within the PDT spectral window at ~690 nm. Photodynamic potential was estimated through DNA cleavage activity of the dpq and dppz complexes in UV-A light of 365 nm and red light of 676 nm. The reactions proceed via the hydroxyl radical pathway. The complexes retain their DNA photocleavage activity in red light under anaerobic conditions, a situation normally prevails in hypoxic tumor core. Investigation into the photocytotoxic potential of these complexes showed that the dppz complex 3 is approximately 4-fold more active in the HEK293 cells expressing human thyrotropin receptor (HEK293-hTSHR) than in the parental cell line and has an insignificant effect on an unrelated human cervical carcinoma cell line (HeLa). Photoexcitation of complex 3 in HEK293-hTSHR cells leads to damage hTSHR as evidenced from the decrease in cAMP formation both in absence and presence of hTSH and decrease in the TSHR immunofluorescence with a concomitant cytoplasmic translocation of the membrane protein, cadherin. The involvement of hTSHR is evidenced from the ability of complex 3 to bind to the extracellular domain of hTSHR (hTSHR-ECD) with a K(d) value of 81 nM and from the photocleavage of hTSHR-ECD.

Casein Kinase CK2 Inhibition Results in Tumor Suppression.

Abstract 5052Protein Kinase II (CK2) is a pleiotropic, and ubiquitous serine/threonine kinase taht utilizes both ATP and GTP as phosphate donors. Protein kinase CK2 mostly exists as a tetramer composed of two catalytic subunits α and α‘, which exists in heterogeneous or homogenous nature, and two regulatory β subunits. CK2 is a key regulator of signaling pathways involved in cell cycle, proliferation and apoptosis. It is consistently overexpressed in cancer tissue and capable of shuttling between cellular compartments but mainly localized in the nuclear matrix of cancer cells. CK2 is highly involved in apoptosis suppression, oncogene activation and tumorigenesis. It is also considered a bad prognostic marker in cancer tissue and is suggested to be a promising target for cancer therapy. In this study, we examined the effect, of a specific protein kinase inhibitor, and ATP competitor, 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole (DMAT), on the growth of various cancer types. We have reported DMAT as a potent cancer therapy. In vitro analysis of the viability of hematologic neoplasms and solid tumors, including human lymphoma U937, hormone dependent breast cancer MCF7, hormone independent breast cancer MDA-MB231 and MDA-MB468, and human cervical cancer HeLa cell lines, revealed marked reduction of cellular viability upon treatment with different DMAT concentration at varying time periods. Lymphoma cell line U937 showed an IC50 between 6 -12 μM. A sharp decrease in cancer cells growth was specifically observed following DMAT treatment of cervical carcinoma HeLa cell line with IC50 between 0.2-0.3 μM. Each of the breast cancer cell lines showed IC50 of 6, 10, and 20 μM DMAT for MDA-MB468, MCF7 and MDA-MB231 respectively. The more cancer cell lines we screen, the more evidence we have to suggest DMAT as a potential anti-cancer therapy. However, the specific mechanism of action of DMAT-inhibited-CK2 pathway in cancer ablation is not clear yet. Using Propidium-Iodide staining in conjunction with flow cytometry techniques, we analyzed and compared cell cycles of treated U937, MCF-7, MDA-MB231, MDA-MB468, and HeLa cell lines. Our data indicate that DMAT induces cell cycle arrest in HeLa cell line at G0/G1 phase. No effect on cell cycle was observed for all other cell lines tested. However, all cell lines underwent apoptosis following treatment with DMAT. Thus far our results suggest that DMAT can induce cell cycle arrest, apoptosis and maybe necrosis or multiple processes at the same time. We suggest that the mechanism of DMAT in cancer inhibition could be of multiple actions which further validate this molecule as a potent cancer therapy that could be suitable for clinical investigation. DisclosuresNo relevant conflicts of interest to declare.

Inhibin/activin-betaE subunit in normal and malignant human cervical tissue and cervical cancer cell lines

Inhibins are dimeric glycoproteins, composed of an alpha-subunit and one of two possible beta-subunits (betaA or betaB), with substantial roles in human reproduction and in endocrine-responsive tumours. Recently a novel beta subunit named betaE was described, although it is still unclear if normal or cancerous cervical epithelial cells as well as cervical cancer cell lines can synthesise the novel inhibin-betaE subunit. About 4 normal cervical tissue samples together with 10 specimens of well-differentiated squamous cervical cancer and adenocarcinoma of the cervix were immunohistochemical analyzed. Additionally, two cervical carcinoma cell lines (HeLa and CaSki) were analyzed by immunofluorescence and RT-PCR for the expression of this novel subunit. We demonstrated for the first time an immunolabelling of the inhibin-betaE subunit in normal and malignant cervical tissue, as well as cervical cancer cells. Although the physiological role is still quite unclear in cervical tissue, inhibin-betaE might play important roles in carcinogenesis. Moreover, the synthesis of this subunit in cervical carcinoma cell lines of squamous and glandular epithelial origins also allows the use of these cell lines in elucidating its functions in cervical cancer pathogenesis. However, since the expression of the inhibin-betaE is minimal in HeLa cells as assessed by immunofluorescence and RT-PCR, the CaSki cell line might be a better model for further functional experiments regarding cervical cancer pathogenesis.

8011 The synergistic effects of nedaplatin and cispaltin on the proliferation and apoptosis of human ovarian carcinoma Skov-3 and cervical carcinoma hela cell line

Ultra-fine grained Al–Fe–Si 8011 alloy was successfully produced by the accumulative roll bonding (ARB) process. However, softening behavior also happened after 2–3 cycles of ARB. The reason of softening behavior was analyzed preliminarily in this paper.

Novel ceramides from aerial parts of Saussurea involucrata Kar. et. Kir.

Novel ceramides, together with nine known compounds were isolated from aerial parts of the Saussurea involucrata Kar. et. Kir. The novel structures were determined by spectroscopic methods and microscale chemical degradation. The ceramides showed appreciable cytotoxicity against three human tumor cell lines including human promyelocytic leukemia (HL-60), human melanoma (A375-S2) and human cervical carcinoma (HeLa) cell lines. The results suggested that this class of ceramides may have potential as an antitumor agent.

Nuclear localization signal peptides enhance transfection efficiency of chitosan/DNA complexes

The purpose of this study was to investigate the potential of nuclear localization signal (NLS) “KPKKKRKV” to mediate the in vitro transfection efficiency of chitosan (CS)/DNA complexes, aiming at its use in gene therapy applications. In the preparation of CS/DNA complexes containing NLS, peptide with NLS was directly incorporated without covalent conjugation to pDNA or chitosan. The gene transfer efficiency of CS/DNA complexes with and without NLS was evaluated in the human cervical carcinoma cell line (Hela cells). The CS/DNA complex containing NLS increased transfection efficiencies in a NLS-dose dependent manner on the Hela cells, compared to the control (CS/DNA complex or NLS). The highest transfection efficiency was significantly observed in CS/DNA complex at the weight ratio of 8 with 120 μg NLS and was 74-fold higher than that in the cells transfected with CS/DNA complex. Cytotoxicity of the NLS/CS/DNA complexes increased as the amount of the peptide increased, however, over 80% average cell viability was observed for complexes at the effective concentration of the peptide for transfections. Therefore, the NLS is expected to be a potent transfection enhancing agent without a covalent conjugation to pDNA or chitosan. Our findings suggest that the high gene expression with the negligible cytotoxicity can be achieved by adding the NLS peptide to chitosan/DNA complexes at an optimal ratio.

Plasmon-induced photothermal cell-killing effect of gold colloidal nanoparticles on epithelial carcinoma cells

Gold colloidal nanoparticles were prepared by the liquid laser ablation of a gold metal plate in water and also by the citrate reduction of HAuCl(4).4H2O. The gold colloidal nanoparticles with the plasmonic band strongly absorb light, which is converted to the photothermal energy. This photothermal energy gives a cytotoxic effect on epithelial carcinoma cells. Interestingly, we found that the size and shape of the nanoparticles are changed by light during the photothermal process in vitro. The cervical carcinoma cell line (HeLa cell) was incubated with the colloidal gold nanoparticles and then exposed to continuous visible light at 400-600 nm with UV- and heat-cutoff filters. The distinct cell-killing effect was observed by this procedure. In the absence of the gold colloidal nanoparticles, only a small amount of cells were photothermally destroyed.

Antitumor Activity of Small Double-Stranded Oligodeoxynucleotides Targeting Telomerase RNA in Malignant Melanoma Cells

Human telomerase RNA (hTR) is an intrinsic component of telomerase enzyme. Small interfering RNAs (siRNAs) and single-stranded antisense oligonucleotides have been used previously for silencing of the hTR. The objective of this study was to investigate the effect of partially double-stranded oligodeoxynucleotides (ODNs), in vitro and in vivo in comparison to single-stranded antisense ODNs and siRNAs. ODNs were designed on the basis of structural properties of an ODN from previous studies on HIV, to target the hTR in the human cervical carcinoma HeLa cell line and mouse telomerase RNA (mTR) in the murine metastatic melanoma B16-F10 cell line, respectively. Our results indicate that ODNs were able to inhibit the hTR by 68% and the mTR by 81% in the respective cell lines. This correlated with ODN-mediated rapid inhibition of cell proliferation and induction of apoptosis excluding slow effects on telomerase function. The inhibition of the hTR was decreased by knock-down of the cellular RNases H suggesting their contribution. Furthermore, we showed a reduction in numbers of metastases by 70% after intravenous administration of ODN-transfected B16-F10 cells in C57BL/6 mice. Our study demonstrates the potential utility of these hairpin-loop-structured ODNs as a different group of nucleic acids for telomerase-based antiproliferative strategies.

The Synergistic Effects of Nedaplatin and Cispaltin on the Proliferation and Apoptosis of Human Ovarian Carcinoma Skov-3 and Cervical Carcinoma Hela Cell Line

The Synergistic Effects of Nedaplatin and Cispaltin on the Proliferation and Apoptosis of Human Ovarian Carcinoma Skov-3 and Cervical Carcinoma Hela Cell Line

Antiproliferation and apoptosis of human tumor cell lines by a lectin (AMML) of Astragalus mongholicus

A lectin (AMML) from the roots of Astragalus mongholicus was extracted and purified by affinity chromatographic technique. Human cervical carcinoma cell line (HeLa), human osteoblast-like cell line (MG63) and human leukemia cell line (K562) were used to check the effects of AMML on cell proliferation, apoptosis and cell cycle. Maximum growth inhibition (92%) was observed with HeLa cells, followed by K562 cells (84%) and MG63 (48%) cells. Morphological observation showed that AMML-treated HeLa cells displayed outstanding apoptosis characteristics, such as nuclear fragmentation and appearance of membrane-enclosed apoptotic bodies. The apoptosis of HeLa cells was confirmed by flow cytometry using Annexin V/FITC and propidium iodide (PI) staining technique. For the first time we also report a significant cell cycle arrest at S phase of HeLa cells by AMML. Therefore, the present investigation may lead to the possible therapeutic use of Astragalus mongholicus lectin.

Early Hematopoietic Zinc Finger Protein Prevents Tumor Cell Recognition by Natural Killer Cells

Early hematopoietic zinc finger/zinc finger protein 521 (EHZF/ZNF521) is a novel zinc finger protein expressed in hematopoietic stem and progenitor cells and is down-regulated during their differentiation. Its transcript is also abundant in some hematopoietic malignancies. Analysis of the changes in the antigenic profile of cells transfected with EHZF cDNA revealed up-regulation of HLA class I cell surface expression. This phenotypic change was associated with an increased level of HLA class I H chain, in absence of detectable changes in the expression of other Ag-processing machinery components. Enhanced resistance of target cells to NK cell-mediated cytotoxicity was induced by enforced expression of EHZF in the cervical carcinoma cell line HeLa and in the B lymphoblastoid cell line IM9. Preincubation of transfected cells with HLA class I Ag-specific mAb restored target cell susceptibility to NK cell-mediated lysis, indicating a specific role for HLA class I Ag up-regulation in the NK resistance induced by EHZF. A potential clinical significance of these findings is further suggested by the inverse correlation between EHZF and MHC class I expression levels, and autologous NK susceptibility of freshly explanted multiple myeloma cells.

Early Hematopoietic Zinc Finger (EHZF/ZNF521) prevents tumor cell recognition by Natural Killer cells. (41.43)

Abstract EHZF/ZNF521 is a novel zinc finger protein expressed in hematopoietic stem and progenitor cells and is downregulated during their differentiation. Its transcript is also abundant in some hematopoietic malignancies. Analysis of the changes in the antigenic profile of cells transfected with EHZF cDNA revealed upregulation of HLA class I cell surface expression. This phenotypic change was associated with increased level of HLA class I heavy chain, in absence of detectable changes in the expression of other antigen processing machinery (APM) components. Enhanced resistance of target cells to NK cell-mediated cytotoxicity was induced by enforced expression of EHZF in the cervical carcinoma cell line HeLa and in the B-lymphoblastoid cell line IM9. Preincubation of transfected cells with HLA class I antigen-specific mAb restored target cell susceptibility to NK cell-mediated lysis, indicating a specific role for HLA class I antigen upregulation in the NK resistance induced by EHZF. A potential clinical significance of these findings is further suggested by the inverse correlation between EHZF and MHC class I expression levels, and autologous NK susceptibility of freshly explanted multiple myeloma cells.

New sesquiterpenes, JBIR-27 and -28, isolated from a tunicate-derived fungus, Penicillium sp. SS080624SCf1

In the course of our screening program for novel metabolites from tunicate-derived fungi, novel sesquiterpenoids, named JBIR-27 (1) and -28 (2), together with known sporogen-AO1 and phomenone, were isolated from the culture broth of Penicillium sp. SS080624SCf1. The structures of 1 and 2 were determined to be eremophilane analogs on the basis of extensive NMR and MS analyses. Sporogen-AO1, phomenone and 2 showed cytotoxicity against human cervical carcinoma cell line HeLa at IC(50) values of 8.3, 19 and 92 microM, respectively, whereas 1 was inactive at a concentration of 80 microM.

Correlativity study between expression of DNA double-strand break repair protein and radiosensitivity of tumor cells

DNA double-strand break (DSB) is generally regarded as the most lethal of all DNA lesions after radiation. Ku80, DNA-PK catalytic subunit (DNA-PKcs) and ataxia telangiectasia mutated (ATM) proteins are major DSB repair proteins. In this study, survival fraction at 2Gy (SF2) values of eight human tumor cell lines (including four human cervical carcinoma cell lines HeLa, SiHa, C33A, Caski, three human breast carcinoma cell lines MCF-7, MDA-MB-231, MDA-MB-453, and one human lung carcinoma cell line A549) were acquired by clone formation assay, and western blot was applied to detect the expressions of Ku80, DNA-PKcs and ATM protein. The correlativity of protein expression with SF2 value was analyzed by Pearson linear correlation analysis. We found that the expression of the same protein in different cell lines and the expression of three proteins in the same cell line had a significant difference. The SF2 values were also different in eight tumor cell lines and there was a positive correlativity between the expression of DNA-PKcs and SF2 (r =0.723, P = 0.043), but Ku80 and ATM expression had no correlation with SF2 (P > 0.05). These findings suggest that the expression level of DNAPKcs protein can be an indicator for predicting the radiosensitivity of tumor cells.

Effects of Res on proliferation and apoptosis of human cervical carcinoma cell lines C33A, SiHa and HeLa

Objective To explore the effects of Res on the proliferation and apoptosis of human cervical cancer cell lines C33A, SiHa and HeLa. Methods Inhibition rates by different concentrations of Res were calculated using MTT method. Apoptosis rates and cell cycles were measured and examined by flow cytometry (FCM). Morphological configuration of apoptotic cells were observed under the fluorescence microscope. Results The growth of cancer cells was inhibited by Res of varied concentrations in a time- and dose-dependent manner ( P<0.01). The cells showed characteristic apoptosis morphologic changes under fluorescence microscope. Res exerted no effects on cell cycles. Conclusion Res inhibits the growth of cervical cell lines C33A, SiHa and HeLa by inducing cell apoptosis in a time- and dose-dependent manner.

Dendritic Effect of Ligand-Coated Nanoparticles: Enhanced Apoptotic Activity of Silica−Berberine Nanoconjugates

We describe the synthesis and biological characterization of a novel prototype, namely, silica nanoconjugates bearing a covalently linked berberine, a plant alkaloid known to have antiproliferative activity. The effect of synthesized nanoconjugates on cell proliferation, the cell cycle profile, and apoptosis in the human cervical carcinoma cell line (HeLa), human hepatocellular liver carcinoma cell line (HepG2), and human embryonic kidney (HEK) 293T cell line has been studied and compared with the results obtained for free berberine. Our results show that all the nanoconjugates display higher antiproliferative activity than free berberine. The ability of these nanoconjugates to inhibit cellular proliferation is mediated by the cell cycle arrest at the G1 phase. Moreover, silica nanoconugates caused selective apoptotic arrest with a higher efficiency than free berberine followed by apoptotic cell death as shown by quantitative fluorescence-activated cell sorting analyses. Efficiency of the nanoconjugates increases upon an increase in the linker chain length, demonstrating the distinct role of the spacer chain that conjugates nanoparticles and ligands. The actual reason to show enhanced efficiency by the nanoconjugates has not been elucidated in the present study; however, we hypothesize that an increase in local concentration due to the confinement of a ligand on the nanosurface ("dendritic" effect) might have led to the observed effect.