Anti-cancer activity of methionine gamma-lyase isolated from Mucor irregularis PQ344458 against cervical adenocarcinoma HeLa cells and pleural lymphoma U937 cells.

Triptolide is a compound extracted from the Chinese herb Tripterygium wilfordii Hook. f. Triptolide has potent anticancer activity. However, the mechanisms by which triptolide exerts its anticancer activities remain unclear. To explore the molecular mechanisms involved in the anticancer activity of triptolide, we have examined the effect of triptolide on the growth of pancreatic carcinoma PANC-1 and cervical adenocarcinoma HeLa cells. We found that treatment of both HeLa and PANC-1 cells with triptolide potently suppressed cell growth and induced apoptosis, indicated by nuclear fragmentation and blebbing. In both HeLa and PANC-1 cells, apoptosis induced by triptolide was associated with activation of both caspase-3 and caspase-8, and cleavage of poly(ADP-ribose) polymerase and Bid. Moreover, in HeLa cells, caspase-9 is also significantly activated in response to triptolide. Overexpression of Bcl-2 in HeLa cells substantially attenuated triptolide-induced apoptosis. Interestingly, substitution of the 14-OH of triptolide with an acetyl group abrogated both its anticancer and its antiinflammatory activities. Our studies suggest that triptolide may exert its anticancer effects by initiating apoptosis through both death-receptor- and mitochondria-mediated pathways. Our results indicate that both the apoptosis-promoting and the antiinflammatory activities of triptolide depend on the 14-OH group.

Triptolide is a compound extracted from the Chinese herb Tripterygium wilfordii Hook F. Triptolide has potent anti-cancer activity. However, the mechanisms by which triptolide exerts its anti-cancer activities remain unclear. To explore the molecular mechanisms involved in the anti-cancer activity of triptolide, we have examined the effect of triptolide on the growth of pancreatic carcinoma PANC-1 and cervical adenocarcinoma HeLa cells. We found that treatment of both HeLa and PANC-1 cells with triptolide potently suppressed cell growth and induced apoptosis, indicated by nuclear fragmentation and blebbing. In both HeLa and PANC-1 cells, apoptosis induced by triptolide was associated with activation of both caspase-3 and caspase-8, and cleavage of PARP and Bid. Moreover, in HeLa cells, capase-9 is also significantly activated in response to triptolide. Over-expression of Bcl-2 in HeLa cells substantially attenuated triptolide-induced apoptosis. Interestingly, substitution of the 14-OH of triptolide with an acetyl group abrogated both its anti-cancer and its anti-inflammatory activities. Our studies suggest that triptolide may exert its anti-cancer effects by initiating apoptosis through both death receptor- and mitochondria-mediated pathways. Our results indicate that both the apoptosis-promoting and the anti-inflammatory activities of triptolide depend on the 14-OH group.

Origanum acutidens (Hand.-Mazz.) Ietsw. is an endemic and perennial plant grown mainly in East Anatolia. Recently, natural plant products have attracted interest due to their safety and therapeutic effects. Therefore, the aim of this study was to investigate phytochemical contents and biological effects of Origanum acutidens. The aerial parts of O. acutidens were extracted with water, ethyl acetate, nbutanol, and methanol/chloroform solvents. Phenolic compounds and other constituents of the extracts were analyzed by HPLC/TOF-MS. The Ethyl Acetate extract (EA) was fractionated by flash chromatography. The extracts and fractions were investigated for their antiproliferative activities on human cervical adenocarcinoma (HeLa) cell line by using BrdU ELISA assay. Antioxidant activities of the extracts and fractions were evaluated by complementary test systems, namely determination of total phenolic contents, metal chelating ability and DPPH radical scavenging assay. Among the extracts, Ethyl Acetate extract (EA) exhibited the highest antiproliferative activity (IC50 = 15.71 ± 0.04 µg/mL) on HeLa cells. It was therefore fractionated by flash chromatography to obtain 10 fractions which were investigated for their phenolic compounds and bioactivities. Rosmarinic acid was determined as the major component of EA and its fractions. EA exhibited higher antiproliferative activity against HeLa cell line than its fractions and 5-fluorouracil (5-FU) at the concentration of 100 µg/mL. EA and its fractions (F10, F6, F4, F7, F3, and F2) displayed higher radical scavenging activity compared to Butylated Hydroxytoluene (BHT). These effects may be attributed to the presence of rosmarinic acid in EA and its active fractions. This study demonstrated that O. acutidens is an essential natural source of polyphenols and a potent natural antioxidant and antiproliferative agent for food and pharmaceutical industries.

Antineoplastic activity of a novel ruthenium complex against human hepatocellular carcinoma (HepG2) and human cervical adenocarcinoma (HeLa) cells

Ionizing radiation induces the production of reactive oxygen species, which play an important causative role in apoptotic cell death. By supplying NADPH for antioxidant systems, we recently demonstrated that the control of mitochondrial redox balance and the cellular defense against oxidative damage are some of the primary functions of mitochondrial NADP(+)-dependent isocitrate dehydrogenase (IDPm). In this study, we demonstrate that modulation of IDPm activity in U937 cells regulates ionizing radiation-induced apoptosis. When we examined the regulatory role of IDPm against ionizing radiation-induced apoptosis in U937 cells transfected with the cDNA for mouse IDPm in sense and antisense orientations, a clear inverse relationship was observed between the amount of IDPm expressed in target cells and their susceptibility to apoptosis. Upon exposure to 2 gray gamma-irradiation, there was a distinct difference between the IDPm transfectant cells in regard to the morphological evidence of apoptosis, DNA fragmentation, cellular redox status, oxidative damage to cells, mitochondrial function, and the modulation of apoptotic marker proteins. In addition, transfection of HeLa cells with an IDPm small interfering RNA decreased the activity of IDPm, enhancing the susceptibility of radiation-induced apoptosis. Taken together, these results indicate that IDPm may play an important role in regulating the apoptosis induced by ionizing radiation, and the effect of IDPm small interfering RNA on HeLa cells offers the possibility of developing a modifier of radiation therapy.

Myelination in Schwann cells is governed by several transcription factors, including the POU proteins Oct6 and Brn2, the high mobility group protein Sox10 and the zinc-finger protein Krox20. How the function of these factors is integrated in the control of myelination has not been established. Previously, we identified an enhancer element controlling Krox20 expression throughout myelination in Schwann cells. In this paper, cell culture experiments were combined with transgenesis to identify transcription factors acting directly upstream of Krox20. The results show that during the promyelin-myelin transition, Krox20 expression is directly activated by Oct6 and Brn2 acting on this enhancer. In addition, the enhancer-dependent synergism between these POU proteins and Sox10 suggests that Krox20 expression requires this combination of factors. These results resolve previous controversy concerning the mechanism of action of Oct6 and Brn2 during myelination and provide an explanation for myelin deficiencies in Waardenberg-Hirschsprung disease patients whereby Sox10 mutations could lead to a loss of Krox20 expression.

The effect of G-protein-coupled estrogen receptor 1 (GPER1) on tumors depends on tumor entity, with its expression level influencing signal transduction and function. Recent research suggests that GPER1 promotes tumor suppression in cervical carcinoma (CC). In contrast, silencing GPER1 increases expression of serpin family E member 1 (SERPINE1) and its protein, plasminogen activator inhibitor-1 (PAI-1), and promotes tumor progression, raising the question of whether PAI-1 might be a suitable target for the treatment of CC. To explore this, we examined the impact of PAI-1 inhibition using Tiplaxtinin (PAI-039, TPX). The effects of TPX treatment on viability, colony formation, migration, and invasion of SiHa cervical squamous cell carcinoma (CSCC) and HeLa cervical adenocarcinoma (CAC) cells were assessed using AlamarBlue, colony formation, gap closure, and Boyden chamber assays, respectively. Apoptosis was examined using the Annexin/PI assay, while the cell cycle was analyzed in more detail using the PI assay. With increasing TPX concentration, viability and colony formation of SiHa and HeLa cells decreased significantly. Cell migration was strongly reduced under PAI-1 inhibitor treatment, while invasion showed a slight decline. Apoptosis and cell cycle were only minimally affected by TPX. PAI-1 inhibitor TPX showed a strong inhibitory effect on both SiHa CSCC and HeLa CAC cells, significantly reducing their viability, colony formation, and migratory capacity. The observed effects suggest that TPX could potentially be used to target and hinder the growth and spread of both CSCC and CAC cells.

Piperine (1-piperoylpeperdine), a nitrogenous pungent substance, is present in the fruits of black pepper (Piper nigrum Linn.) and long pepper (Piper longum Linn.). It possesses several pharmacological properties and has been extensively explored for its anti-cancerous activities. The mechanism underlying its anti-cancer potential in human cervical adenocarcinoma (HeLa) cells is not well interpreted. The anti-proliferative effect and the mode of action of piperine were investigated through some potent markers of apoptosis viz.reactive oxygen species (ROS) generation, cellular apoptosis and loss of mitochondrial membrane potential (MMP). DNA fragmentation, cell cycle kinetics, caspase-3 activity and cell migration assays were also conducted to observe the efficacy of piperine against HeLa cells. The results showed that piperine exposure induces apoptosis significantly in a dose-dependent manner and inhibits the growth of HeLa cells with an increase in ROS generation, nuclear condensation and delayed wound healing. In addition, piperine also encourages cell death by the loss of MMP, DNA fragmentation and the activation of caspase-3. Growth inhibition of HeLa cells was found to be associated with G2/M phase arrest and sub-G1 accumulation. The present study provides useful insight into the apoptotic potential of piperine and further in vivo and clinical studies will be needed for its validation and in the finding of more effective and least toxic regimens against cervical cancer.

Objective: In the current study, an extract of turmeric rhizome (Turmesac®) was evaluated for possible anticancer activity in human cervical adenocarcinoma (HeLa) cells.
 Methods: Turmesac®’s ability to elicit cytotoxicity in cancer cells was evaluated by (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay (MTT) assay, where the IC50 value was determined. Apoptosis induction and cell cycle arrest were analyzed by flow cytometry post-Turmesac® IC50 value treatment for 24 h.
 Results: The determined IC50 value of Turmesac® in HeLa cells was 115.12 μg/ml. This concentration was able to induce apoptosis 2 times greater than the apoptotic standard, camptothecin, treated cells. Cell cycle arrest was observed at the G0/G1 and S phases in Turmesac® treated HeLa cells.
 Conclusion: Turmesac® shows the potential of being a promising anticancer agent that may be incorporated into chemotherapies, but further study is required to elucidate the exact mechanisms involved with longer treatment duration, as would be the case in clinical trial phases.

Taking advantage of the fluorescence property of alkyne-modified naphthalimide, the two-photon excited fluorogenic probe Naph-yne is used for azide-tagged mannosyl glycoprotein imaging both at the cellular level and for vertebrate model organisms.

DNA interaction, antimicrobial, antioxidant and anticancer studies on Cu(II) complexes of Luotonin A

This study was performed to determine whether relaxin influences the growth of human adenocarcinoma (HeLa) cells of the cervix, which has been shown to contain relaxin-binding sites and proliferate by relaxin. Cells were maintained in Dulbecco's modified Eagle's medium with 5% fetal calf serum. Highly purified porcine relaxin was added at concentrations of 10(-11) to 10(-5) M and incubated for 44, 69, 94, and 118 h. Cell proliferation/cytotoxicity and endogenous nitric oxide production were determined using a colorimetric MTS assay and Griess assay, respectively. The results indicate that whereas relaxin promotes the growth rate of MCF-7 cells, which is in agreement with previous reports, at the doses and times of exposure used in this study relaxin does not significantly influence the number of viable HeLa cells. However, relaxin at nanomolar concentrations (10(-9) to 10(-8) M) promoted the growth of HeLa cells in the presence of both estrogen and progesterone to a small extent. In agreement with these results, relaxin did not affect nitric oxide production. We conclude that relaxin may have differential effects on the growth of two different cancer cell types, MCF-7 and HeLa cells.

We have previously described the isolation of a replication competent (RC) complex from calf thymus, containing DNA polymerase alpha, DNA polymerase delta and replication factor C. Here, we describe the isolation of the RC complex from nuclear extracts of synchronized HeLa cells, which contains DNA replication proteins associated with cell-cycle regulation factors like cyclin A, cyclin B1, Cdk2 and Cdk1. In addition, it contains a kinase activity and DNA polymerase activities able to switch from a distributive to a processive mode of DNA synthesis, which is dependent on proliferating cell nuclear antigen. In vivo cross-linking of proteins to DNA in synchronized HeLa cells demonstrates the association of this complex to chromatin. We show a dynamic association of cyclins/Cdks with the RC complex during the cell cycle. Indeed, cyclin A and Cdk2 associated with the complex in S phase, and cyclin B1 and Cdk1 were present exclusively in G(2)/M phase, suggesting that the activity, as well the localization, of the RC complex might be regulated by specific cyclin/Cdk complexes.

ENOX2 (tNOX), a tumor-associated cell surface ubiquinol (NADH) oxidase, functions as an alternative terminal oxidase for plasma membrane electron transport. Ubiquitous in all cancer cell lines studied thus far, ENOX2 expression correlates with the abnormal growth and division associated with the malignant phenotype. ENOX2 has been proposed as the cellular target for various quinone site inhibitors that demonstrate anticancer activity such as the green tea constituent epigallocatechin-3-gallate (EGCg) and the isoflavone phenoxodiol (PXD). Here we present a possible mechanism that explains how these substances result in apoptosis in cancer cells by ENOX2-mediated alterations of cytosolic amounts of NAD(+) and NADH. When ENOX2 is inhibited, plasma membrane electron transport is diminished, and cytosolic NADH accumulates. We show in HeLa cells that NADH levels modulate the activities of two pivotal enzymes of sphingolipid metabolism: sphingosine kinase 1 (SK1) and neutral sphingomyelinase (nSMase). Their respective products sphingosine 1-phosphate (S1P) and ceramide (Cer) are key determinants of cell fate. S1P promotes cell survival and Cer promotes apoptosis. Using plasma membranes isolated from cervical adenocarcinoma (HeLa) cells as well as purified proteins of both bacterial and human origin, we demonstrate that NADH inhibits SK1 and stimulates nSMase, while NAD(+) inhibits nSMase and has no effect on SK1. Additionally, intact HeLa cells treated with ENOX2 inhibitors exhibit an increase in Cer and a decrease in S1P. Treatments that stimulate cytosolic NADH production potentiate the antiproliferative effects of ENOX2 inhibitors while those that attenuate NADH production or stimulate plasma membrane electron transport confer a survival advantage.

Cancer is one of the leading causes of death in the world affecting millions of people. The commercially available anticancer drugs lack the selectivity and show several undue side effects during the biologically targeted therapy, thus calling for the exploration of wider chemical space to furnish new structural leads with promising anticancer potential. In this endeavor, we synthesized a series of coumarinyl thiazolotriazoles with diverse functional group tolerance and will be tested for their anticancer properties against cancer cell lines (HeLa and MCF-7) and a normal cell line (BHK-21). To overcome such complications, in the current study, we evaluated the cytotoxic effects of coumarinyl thiazolotriazole hybrids on human breast adenocarcinoma (MCF-7), cervical adenocarcinoma (HeLa) cells and normal cells i.e., Baby Hamster Kidney cells (BHK-21) using MTT (dimethyl-2-thiazolyl- 2,5-diphenyl-2H-tetrazolium bromide) assay. DNA binding studies of compound 6c was performed on Herring- Sperm DNA (HS-DNA) and docking studies were also carried out. The mechanistic studies were performed on potent compounds by fluorescent microscopic studies, release of Lactate Dehydrogenase (LDH) and mitochondrial membrane potential, activation of caspase-9 and -3 and flow cytometric analysis. As revealed by MTT assay, compounds 6m and 6c were identified as the most potent derivatives among the tested series with IC50 values of 5.64 and 29.1 μM against HeLa and MCF cells, respectively as compared to cisplatin which gave IC50 values of 11.3 and 6.20 μM, respectively. DNA binding studies of compound 6c showed the binding of compound in DNA with Gibbs free energy of ‒17 KJ/mol and docking studies validated the DNA binding studies. Fluorescent microscopic studies using 4',6-diamidino-2-phenylindole (DAPI) and Propidium Iodide (PI) staining confirmed the occurrence of apoptosis in HeLa cells treated with the most active compound 6m. Moreover, compounds 6m and 6c also triggered the release of Lactate Dehydrogenase (LDH) in treated HeLa and MCF-7 cells while a luminescence assay displayed a remarkable increase in the activity of caspase-9 and -3. Moreover, flow cytometric results revealed that compound 6m caused G0 /G1 arrest in the treated HeLa cells. Our results suggested that the compound possesses chemotherapeutic properties against breast cancer and cervical adenocarcinoma cells, thus warranting further research to test the anticancer efficacy of this compound at clinical level.