Mammalian Cancer Cell Lines Research Articles

MicroRNA modified tumor-derived exosomes as novel tools for maturation of dendritic cells.

Tumor-derived exosomes (TEX) are known by their immune suppression effects as well as initiation mediators in cancer progression and metastasis. Meanwhile, they are appropriate sources to induce immunity against tumor cells, as consist of tumor specific and associated antigens. The aim of the current study is modifying TEX with microRNA miR-155, miR-142, and let-7i, to enhance their immune stimulation ability and induce potent dendritic cells (DC). For this, exosomes were isolated from mouse mammalian breast cancer cell line; 4T1, and subjected to miR-155, miR-142, and let-7i by electroporation. Immature DCs were generated from mouse bone marrow in the presence of interleukin-4 (IL-4) and granulocyte-macrophage colony-stimulating factor (GM-CSF). To mature DCs, lipopolysaccharide (LPS), TEX, and modified TEX were used. The expression level of miRNAs and their target genes (IL-6, IL-17, IL-1b, TGFβ, SOCS1, KLRK1, IFNγ, and TLR4) was determined. TEX were nanovesicles with spheroid morphology which expressed CD81, CD63, and TSG101, as exosome markers, at protein level. MHCII, CD80, and CD40 as maturation markers were assessed by flow cytometry. Overexpression of miRNAs were confirmed in exosomes and mDCs. Up and downregulation of target genes confirmed the gene network in DC maturation. We found that Let-7i could efficiently induce the DC maturation, as well as miR-142 and miR-155 have enhancing effects. These findings reveal that the modified TEX would be a hopeful cell-free vaccine for the cancer treatment.

ANTI-PROLIFERATIVE ACTIVITIES OF THE AQUEOUS ROOT EXTRACT OF DIANTHUS THUNBERGII SS HOOPER (CARYOPHYLLACEAE)

Background: The roots of Dianthus thunbergii SS Hooper are used traditionally in South Africa for the treatment of
 diabetes, wounds, colic, chest complaints and cancer. This study was aimed at investigating the potential anti-proliferative
 activities of the D. thunbergii in mammalian cancer cell lines.
 Materials and Methods: Aqueous and ethanol extracts of D. thunbergii were tested in vitro on two cancer cell lines:
 human hepato-cellular carcinoma (HepG2) cells and murine insulinoma (INS-1) cells using the 3-(4,5-Dimethylthiazol-2-
 yl) 2,5-diphenyltetrazolium bromide (MTT) and crystal violet cell viability assays, as well as live-cell fluorescence imaging
 microscopy. A tentative profiling of the aqueous extract was also carried out using liquid chromatography-mass
 spectrometry (LC-MS).
 Results: The aqueous extract (50-200μg/ml) exhibited significant (p

Phenylselenyl containing turn-on dibodipy probe for selective detection of superoxide in mammalian breast cancer cell line

A monoselenium containing diBODIPY-based probe was synthesized. The probe was found to be selective and sensitive for the detection of superoxide (KO2) over other reactive oxygen species through fluorescence turn-on event with 11-fold increase in fluorescence intensity. The detection limit was found to be 4.42 μM. The utility of the probe for detection of intracellular superoxide was demonstrated in a mammalian breast cancer cell line (MCF-7) through fluorescence microscopy.

Plant thionins: structure, biological functions and potential use in biotechnology

Antimicrobial peptides (AMPs) are important components of defense system in both plants and animals. They represent an ancient mechanism of innate immunity providing rapid first line of defense against pathogens. Plant AMPs are classified into several families: thionins, defensins, nonspecific lipid-transfer proteins, hevein- and knottin-type peptides, hairpinins and macrocyclic peptides (cyclotides). The review focuses on the thionin family. Thionins comprise a plant-specific AMP family that consists of short (~5 kDA) cysteine-rich peptides containing 6 or 8 cysteine residues with antimicrobial and toxic properties. Based on similarity in amino acid sequences and the arrangement of disulphide bonds, five structural classes of thionins are discriminated. The three-dimensional structures of a number of thionins were determined. The amphipathic thionin molecule resembles the Greek letter Г, in which the long arm is formed by two antiparallel α-helices, while the short one, by two parallel β-strands. The residues responsible for the antimicrobial activity of thionins were identified. Thionins are synthesized as precursor proteins consisting of a signal peptide, the mature peptide region and the C-terminal prodomain. Thionins protect plants from pathogenic bacteria and fungi acting directly on the membranes of microorganisms at micromolar concentrations, although their precise mode of action remains unclear. In addition to plant pathogens, thionins inhibit growth of a number of human pathogens and opportunistic microorganisms, such as Candida spp., Saccharomyces cerevisiae, Fusarium solani, Staphylococcus aureus and Escherichia coli. Thionins are toxic to different types of cells including mammalian cancer cell lines. Transgenic plants expressing thionin genes display enhanced resistance to pathogens. A wide range of biological activities makes thionins promising candidates for practical application in agriculture and medicine.

Anti-tumor activity of Escherichia coli Shiga toxin A subunit delivered by SF9 insect cells

Cancer remains a major health problem around the world. A Shiga toxin is a bacterial toxin often produced by Shigella dysenteriae and Escherichia coli. A subunit of the Shiga toxin (StxA) is a cytotoxic agent which could be used to induce death in cancer cells.StxA expressed from baculovirus was evaluated in a pTriEx™ expression vector. The baculovirus vector was used for the A subunit delivery of StxA. StxA cell cytotoxicity was induced by the virus and assessed in the MCF7 and HeLa cell lines. In addition, the breast cancer cytotoxicity of the expressed StxA was also assessed in a cancer induced in mice.The cytotoxicity of the recombinant StxA baculovirus with different multiplicities of infection (MOI) was measured. The results showed that significant cytotoxicity can be induced on the mammalian epithelial breast cancer cell lines, MCF7 and HeLa cells with MOI ≥ 2. The results also showed that a malignant tumor induced by MCF7 could be inhibited in a mouse cancer model.Therefore, it can be concluded that StxA, expressed by baculovirus, could be used for in vitro and in vivo gene delivery. In this study StxA, delivered by the baculovirus inhibited cell proliferation, and eliminated HeLa and MCF7 cells, in vitro. In conclusion, this method can be used as a safe alternative for anticancer drug delivery inside cancer cells.

Metabolomic Analysis of HER2‐positive Breast Cancer Cells

PurposeHER2‐positive breast cancer accounts for more than 20% of the diagnosed cases and is characterized by aggressive growth, increased disease recurrence, and poor prognosis. While much of the signaling mechanism(s) have been elucidated, the impact of HER2 positivity on cellular metabolism is not well understood. We examined (1) gene expression profiles of mammalian breast cancer cell lines with increasing HER2 expression levels (MCF10A, MDA‐MB‐468, MDA‐MB‐453, HCC1954, HCC2218) and correlated changes to metabolic expression profiles and (2) the effect of serum starvation and pervanadate treatment on the metabolic profiles of these HER2‐expressing cell lines.MethodsOur study used a combination of 1H‐NMR and 2D‐NMR (TOCSY), multivariate analyses, and protein expression validation in human breast cancer cell lines (untreated, serum‐starved, serum‐starved with pervanadate treatment).ResultsWe found that both gene and metabolic expression profiles of HER2‐positive cell lines correlated with leucine and isoleucine biosynthesis (p‐value < 0.05). Comparative analyses of profiles from untreated, serum‐starved, and serum‐starved with pervanadate‐treated cell samples revealed that lactate concentration (p‐value <0.05) dropped significantly with serum starvation and pervanadate treatment consistent with reduced glycolytic metabolism and potentially mediated through suppression of tyrosine phosphatase activity.ConclusionThis study highlights important metabolites that are potential biomarkers of HER2‐positive breast cancer and suggests alternate/novel pathways to consider for future understanding and treatment.Support or Funding InformationThis work was funded in part through intramural programs at Fresno State to MKS and JAB, and through the NIGMS 1SC3GM122630‐01 to JAB.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Polycyclic polyprenylated acylphloroglucinols and biphenyl derivatives from the roots of Garcinia nuntasaenii Ngerns. & Suddee

Seven previously undescribed compounds, including three polycyclic polyprenylated acylphloroglucinols (garcinuntins A–C), three biphenyl derivatives (garcinuntabiphenyls A–C) and a lanostane triterpene (garcinuntine), along with thirteen known compounds were isolated from the root of Garcinia nuntasaenii Ngerns. & Suddee. Their structures were elucidated on the basis of spectroscopic techniques. For garcinuntins A–C, the absolute configurations were confirmed by the combination of single X-ray crystallography and ECD calculations. Anti-HIV activity using anti-HIV-1 reverse transcriptase and syncytium inhibition assays, and cytotoxic activity against a panel of cultured mammalian cancer cell lines of isolated compounds were investigated.

Substrates with patterned topography reveal metastasis of human cancer cells

In this study, we aimed at studying the effects of engineered and patterned substrates on the migration characteristics of mammalian cancer cell lines. On the shallow topographical patterns, cells from different histological origins showed different migration speed and directionality. We also observed that cells from the same origin showed distinctive behaviour, suggesting these substrate topographies could distinguish cancer subtypes. To eliminate the influence of genetic background, we examined two isogenic subpopulations of ovarian cancer cell lines for their different metastatic activities. While these cell lines showed indistinguishable migration characteristics on a flat substrate, their motilities on the patterned substrates were highly different, suggesting that cancer cells’ motilities on these substrates varied in a metastasis-dependent manner. While cells with different metastatic activities showed similar morphology and focal adhesion distribution on flat surface, vinculin aggregated into single cytoplasmic foci in metastatic cells cultured on the engineered substrates. This implies that the topographical patterns on the substrates induced vinculin redistribution in cancer cells with a higher invasive activity. The fabricated platforms with topographical patterns offer a novel in vitro technique for metastasis assessment. Moreover, such platforms could potentially provide the opportunity to sort cells in different metastatic states using advanced pattern designs and features.

Environmental cystine drives glutamine anaplerosis and sensitizes cancer cells to glutaminase inhibition.

Many mammalian cancer cell lines depend on glutamine as a major tri-carboxylic acid (TCA) cycle anaplerotic substrate to support proliferation. However, some cell lines that depend on glutamine anaplerosis in culture rely less on glutamine catabolism to proliferate in vivo. We sought to understand the environmental differences that cause differential dependence on glutamine for anaplerosis. We find that cells cultured in adult bovine serum, which better reflects nutrients available to cells in vivo, exhibit decreased glutamine catabolism and reduced reliance on glutamine anaplerosis compared to cells cultured in standard tissue culture conditions. We find that levels of a single nutrient, cystine, accounts for the differential dependence on glutamine in these different environmental contexts. Further, we show that cystine levels dictate glutamine dependence via the cystine/glutamate antiporter xCT/SLC7A11. Thus, xCT/SLC7A11 expression, in conjunction with environmental cystine, is necessary and sufficient to increase glutamine catabolism, defining important determinants of glutamine anaplerosis and glutaminase dependence in cancer.

Synthesis, structure, magnetic and biological activity studies of bis-hydrazone derived Cu(ii) and Co(ii) coordination compounds.

Four coordination compounds of formulae [Cu(II)2(H2L(1))(HL(1))](ClO4)3·H2O (1), [Cu(II)2(H2L(2))(CH3OH)2](ClO4)2·2CH3OH (2), [Co(II)2(H2L(1))2](ClO4)4 (3) and [Co(II)2(H2L(2))2]·2H2O (4) were synthesized via self-assembly of succinohydrazone derived ligands (H2L(1) = N',N'-4-bis(2-pyridyl)succinohydrazide, H4L(2) = N',N'-4-bis(2-hydroxybenzylidene)succinohydrazide) and Cu(2+) and Co(2+) ions, respectively. The compounds were characterized by crystal structure determination, magnetic measurements and biological activities. Compounds 1, 3 and 4 have discrete double helicate structures, whereas compound 2 is a one-dimensional chain. Magnetic studies show antiferromagnetic exchange interactions in 2 with a J value of -67.1 cm(-1) and antiferromagnetic spin-canting in compound 3 originates through supramolecular H-bonding. For compound 3, a clear bifurcation was observed in zero field cooled (ZFC) and field cooled (FC) measurement at a temperature of 3.5 K and field of 0.1 T, implying long range magnetic ordering below this temperature. Interestingly, all of compounds 1-4 show significant changes in their absorption (hypo- and hyperchromism) in the presence of SS-DNA, inferring interaction between the compounds and DNA. In addition, compounds 1-4 significantly exhibited nuclease activities on both RNA and pUC19 plasmid DNA. Moreover, the nuclease activity was further enhanced in the presence of oxidant (H2O2) and suggests the possible role of reactive oxygen species in DNA nicking ability of compounds 1-4. Furthermore, compounds 1, 2 and 4 exhibited significant cytotoxicity against mammalian cancer cell lines (HeLa, A549 and MDAMB-231). In addition, our results from Annexin/PI staining and DNA fragmentation assays revealed that these compounds are capable of inducing apoptosis and have potential to act as anticancer drugs.

Novel series of coumarinyl substituted-thiazolidin-2,4-dione analogs as anticancer agents: design, synthesis, spectral studies and cytotoxicity evaluation.

In this research work, a series of eighteen novel coumarinyl substituted thiazolidin-2,4-dione analogs (4a-4r) have been designed by molecular hybridization approach, synthesized and their structures were established on the basis of FTIR, 1H NMR, 13C NMR and elemental (CHN) analysis. These title compounds were screened for their cytotoxicity using MTT assay methodology against five different mammalian cancer cell lines viz. hormone dependant breast adenocarcinoma (MCF7), cervical carcinoma (HeLa), colorectal carcinoma (HT29), lung cancer (A549) and prostate adeno carcinoma (PC3). The cytotoxicity screening studies revealed that MCF-7, HeLa and A549 cancer cell lines were sensitive to all the tested compounds. Though the compounds showed varying degrees of cytotoxicity in the tested cell lines, most significant effect was observed for compounds 4i (1.06, 2.4 and 3.06 µM) and 4o (0.95, 3.2 and 2.38 μM) against MCF7, HeLa and A549 cell lines respectively. In conclusion, the anticancer results of these promising leads strongly encouraged us for additional lead optimization with the aim of developing more potential anticancer agents.

Design, synthesis and anticancer activity of functionalized spiro-quinolines with barbituric and thiobarbituric acids

A new series of spiro-quinoline compounds have been accomplished by the reaction of barbituric acid or thiobarbituric acid with derivatives of benzisoxazole-5-carbaldehyde or 2-substituted benzaldehyde. These compounds were evaluated for their in vitro cytotoxicity on two mammalian cancer cell lines MCF-7 and KB. The compounds exhibit cytotoxicity against these cell lines in micromolar range. Among the series of compounds, 11(a–j) particularly 11b and 11e showed relatively good activity against both the tested cell lines. Compound 11b was found to exhibit the highest cytotoxic activity with IC50 value 90.2 µM for MCF-7 and 49.8 µM for KB cell line. Flow cytometric analysis study confirmed that these molecules induced cytotoxicity via apoptosis.

Alteration in global DNA methylation after bisphenol a exposure in MCF-7 cells

Abstract: Bisphenol A (BPA), as synthetic monomer used especially in the production of polycarbonate plastic and epoxy resins, has endocrine disruptor properties and high risk on human health. Continuous release of free BPA into food, beverages, and environment has resulted in a widespread human exposure to this chemical. Role of endocrine effects of environmental chemicals such as BPA on the changes in gene expression may be associated with epigenetic mechanisms. Early detection of aberrant DNA methylation, which regulates transcription of tumor suppressor genes and activation of oncogenes in carcinogenesis process, may provide powerful mechanistic insights in the toxicity of BPA. 5-hydroxymethylcytosine (5-hmC), oxidation product of 5-methylcytosine (5-mC), is considered as a new epigenetic DNA modification with relevant roles in cell homeostasis regulating DNA demethylation and transcription. Our aim was to investigate possible changes in the global 5-mC and 5-hmC after 48 and 96 h BPA exposure in mammalian breast cancer cell line (MCF-7). 50% of inhibitory concentration (IC50) value of BPA was determined as 148 μM by MTT cytotoxicity test in MCF-7 cells. We revealed decrease on the level of global 5-mC after BPA exposure (100 nM and 1 μM) for 48 and 96 h by using 5-mC Elisa kit, whereas non-significant slightly increase were observed in the levels of 5-hmC%. We suggested that global DNA methylation may be involved in BPA toxicity in breast cancer cells. Furthermore, evaluation on the expression of enzymes involved in DNA methylation and demethylation after BPA exposure should be investigated in cell cultures. Key words: Bisphenol A, 5-methylcytosine, 5-hydroxymethylcytosine, Cytotoxicity, MCF-7 cells.

Abstract 4341: Metabolomic analysis of HER2-positive breast cancer cells

Abstract Purpose: HER2-positive breast cancer accounts for more than 20% of the diagnosed cases and is characterized by aggressive growth, increased disease recurrence, and poor prognosis. While much of the signaling mechanism(s) have been elucidated, the impact of HER2 positivity on cellular metabolism is not well understood. We examined (1) gene expression profiles of mammalian breast cancer cell lines with increasing HER2 expression levels (MCF10A, MDA-MB-468, MDA-MB-453, HCC1954, HCC2218) and correlated changes to metabolic expression profiles and (2) the effect of serum starvation and pervanadate treatment on the metabolic profiles of these HER2-expressing cell lines. Methods: Our study used a combination of 1H-NMR and 2D-NMR (TOCSY), bioinformatics analyses, and protein expression validation in human breast cancer cell lines (untreated, serum-starved, serum-starved with pervanadate treatment). Results: We found that both gene and metabolic expression profiles of HER2-positive cell lines correlated with leucine and isoleucine biosynthesis (p-value < 0.05). Comparative analyses of profiles from untreated, serum-starved, and serum-starved with pervanadate-treated cell samples revealed that lactate concentration (p-value <0.05) dropped significantly with serum starvation and pervanadate treatment consistent with reduced glycolytic metabolism and potentially mediated through suppression of tyrosine phosphatase activity. Conclusion: This study highlights important new metabolites that are potential biomarkers of HER2-positive breast cancer and suggests alternate/novel pathways to consider for future understanding and treatment. Citation Format: Malika K. Sahni, Resmi K. Ravindran, Imran H. Khan, Jason A. Bush, V. V. Krishnan. Metabolomic analysis of HER2-positive breast cancer cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4341. doi:10.1158/1538-7445.AM2014-4341

Abstract 3479: The cellular thermal shift assay as a tool for setting indivual treatment schemes

Abstract Many cancers respond insufficiently to existing therapies and acquired drug resistance often develops. To address this challenge, a range of different therapeutic strategies is currently in development. Many of these are based on the concept of targeted therapies, where target proteins are drivers of cancer processes or synthetic vulnerabilities of cancer cells. In this project we have used the newly developed CETSA method to monitor levels of drug binding to specific target proteins in tumors and primary cells from breast cancer patients. CETSA is a generic method for evaluating drug binding to target proteins in cells and tissues. The technique is based on the physical phenomenon of ligand-induced thermal stabilization of target proteins. Using this technique, it is possible to look at interactions between a drug and protein of interest without having to label them. Previously, we have validated drug binding in mammalian cancer cell lines for a set of important clinical targets and shown that a range of critical factors that are important for drug development can be identified at the target engagement level, including drug transport and activation, off-target effects, drug resistance as well as drug distribution in animal tissues. By using different drugs and amounts we now have preliminary data suggesting that CETSA could be used as a tool for optimizing treatment schemes for individual care and for detecting resistance development at an early stage. Citation Format: Daniel M. Molina, Pär Nordlund. The cellular thermal shift assay as a tool for setting indivual treatment schemes. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3479. doi:10.1158/1538-7445.AM2014-3479

Investigation of 6-[¹⁸F]-fluoromaltose as a novel PET tracer for imaging bacterial infection.

Despite advances in the field of nuclear medicine, the imaging of bacterial infections has remained a challenge. The existing reagents suffer from poor sensitivity and specificity. In this study we investigate the potential of a novel PET (positron emission tomography) tracer that overcomes these limitations.Methods6-[18F]-fluoromaltose was synthesized. Its behavior in vitro was evaluated in bacterial and mammalian cultures. Detailed pharmacokinetic and biodistribution profiles for the tracer were obtained from a murine model.Results6-[18F]-fluoromaltose is taken up by multiple strains of pathogenic bacteria. It is not taken up by mammalian cancer cell lines. 6-[18F]-fluoromaltose is retained in infected muscles in a murine model of bacterial myositis. It does not accumulate in inflamed tissue.ConclusionWe have shown that 6-[18F]-fluoromaltose can be used to image bacterial infection in vivo with high specificity. We believe that this class of agents will have a significant impact on the clinical management of patients.

Rapid one-step purification of single-cells encapsulated in alginate microcapsules from oil to aqueous phase using a hydrophobic filter paper: implications for single-cell experiments.

By virtue of the biocompatibility and physical properties of hydrogel, picoliter-sized hydrogel microcapsules have been considered to be a biometric signature containing several features similar to that of encapsulated single cells, including phenotype, viability, and intracellular content. To maximize the experimental potential of encapsulating cells in hydrogel microcapsules, a method that enables efficient hydrogel microcapsule purification from oil is necessary. Current methods based on centrifugation for the conventional stepwise rinsing of oil, are slow and laborious and decrease the monodispersity and yield of the recovered hydrogel microcapsules. To remedy these shortcomings we have developed a simple one-step method to purify alginate microcapsules, containing a single live cell, from oil to aqueous phase. This method employs oil impregnation using a commercially available hydrophobic filter paper without multistep centrifugal purification and complicated microchannel networks. The oil-suspended alginate microcapsules encapsulating single cells from mammalian cancer cell lines (MCF-7, HepG2, and U937) and microorganisms (Chlorella vulgaris) were successfully exchanged to cell culture media by quick (~10 min) depletion of the surrounding oil phase without coalescence of neighboring microcapsules. Cell proliferation and high integrity of the microcapsules were also demonstrated by long-term incubation of microcapsules containing a single live cell. We expect that this method for the simple and rapid purification of encapsulated single-cell microcapsules will attain widespread adoption, assisting cell biologists and clinicians in the development of single-cell experiments.

Isolation and synthesis of N-acyladenine and adenosine alkaloids from a southern Australian marine sponge, Phoriospongia sp.

Chemical fractionation of the southern Australian marine sponge Phoriospongia sp. (CMB-03107) yielded phorioadenine A (1) as a nematocidal agent and the first reported example of a 6-N-acyladenine natural product. The structure of 1 was confirmed by spectroscopic analysis and the chemical synthesis of racemic (1a) and enantiomeric (1b) analogues. HPLC–ESIMS analysis of the crude sponge extract with comparisons to the synthetic 6-N-acyladenosine 2a provided evidence that the biosynthetically related adenosine, phorioadenosine A (2), was present as a trace co-metabolite. The rare starfish metabolite asterubine (3) was also isolated as a co-metabolite, and its structure confirmed by spectroscopic analysis and chemical synthesis. Biological investigations confirmed that natural products 1–3 and synthetic analogues 1a–e and 2a were not cytotoxic to multiple mammalian cancer cell lines, or Gram-positive or -negative bacteria. Nematocidal activity (inhibition of larval development of Haemonchus contortus) detected in the Phoriospongia sp. extract was attributed to 1 (LD99 31μg/mL), with preliminary structure–activity relationship investigations confirming the importance of the N-acyl side chain.

STAT3 and MCL-1 associate to cause a mesenchymal epithelial transition

Embryo implantation is effected by a myriad of signaling cascades acting on the embryo-endometrium axis. Here we show, by using MALDI TOF analysis, far-western analysis and colocalization and co-transfection studies, that STAT3 and MCL-1 are interacting partners during embryo implantation. We show in vitro that the interaction between the two endogenous proteins is strongly regulated by estrogen and progesterone. Implantation, pregnancy and embryogenesis are distinct from any other process in the body, with extensive, but controlled, proliferation, cell migration, apoptosis, cell invasion and differentiation. Cellular plasticity is vital during the early stages of development for morphogenesis and organ homeostasis, effecting the epithelial to mesenchymal transition (EMT) and, the reverse process, mesenchymal to epithelial transition (MET). STAT3 functionally associates with MCL-1 in the mammalian breast cancer cell line MCF7 that overexpresses STAT3 and MCL-1, which leads to an increased rate of apoptosis and decreased cellular invasion, disrupting the EMT. Association of MCL-1 with STAT3 modulates the normal, anti-apoptotic, activity of MCL-1, resulting in pro-apoptotic effects. Studying the impact of the association of STAT3 with MCL-1 on MET could lead to an enhanced understanding of pregnancy and infertility, and also metastatic tumors.

Biological evaluation of 8-alkyl xanthines as potential cytotoxic agents

A series of 8-substituted alkyl xanthines were evaluated in vitro to test the cytotoxocity in cells. For this experiment, we utilized different mammalian cancer cell lines primarily representing prostrate and lung. One of the compounds synthesized, viz. 8-tertbutyl caffeine showed potent anticancer activity at low concentrations against DU145 when compared to adriamycin. Further experiments were carried out to check the cell cycle arrest in the DU145 cells treated with adriamycin, caffeine and 8-tert butyl caffeine. We observed that there was an arrest in G1 phase of cell cycle at 24 hours while at 48 hours of incubation, the cells were constantly distributed (59.71% -70.79%). We conclude that the effect of 8-tertbutyl caffeine is relatively comparable to caffeine whereas in adriamycin treated cells, we observed the cells underwent G2 arrest. We evaluate the studies on these effects by showing potent analogues which could be used as promising anticancer agents.