Normal Human Lung Cell Line Research Articles

Synthesis, characterization, antiproliferative activity, docking, and molecular dynamics simulation of new 1,3-dihydro-2H-benzimidazol-2-one derivatives

Cancer is a global public health problem that affects millions each year. Novel anticancer drug candidates are in need to treat various cancers and to overcome the resistance that exists against drugs in use. Benzimidazole derivatives have been reported as anticancer agents. These lead us to synthesize similar benzimidazole derivatives and investigate their anticancer activity. In this study, six new 1,3-dihydro-2H-benzimidazol-2-one-based molecules (2a-f) were synthesized. The structures of these molecules were verified by spectroscopic methods. The antiproliferative activities of molecules 2a-f were screened against a panel of human cancer cell lines, including the liver, colon, lung, and breast. The molecules were also tested towards normal human lung cell line to determine their selectivity. The results demonstrated that compound 2d had the highest cytotoxic effect compared to compounds 2a-c, 2e, and 2f against DLD-1 and MDA-MB-231 cell lines. The binding potential of the relatively active compound, 2d, with three targets was investigated through molecular docking. The stability of target-compound complexes procured from the docking was explored through molecular dynamics (MD) simulation. The docking and MD simulation studies revealed that compound 2d had the highest potential to bind to GALR3 among the targets. Furthermore, the computational pharmacokinetic study demonstrated that the synthesized compounds had drug-like properties. Communicated by Ramaswamy H. Sarma

Molecular studies regarding colon cancer cell line (HCT116) treated by new ruthenium and palladium complexes

Ru(H2dtpa) (H2O) and Pd (DME)Cl2 were prepared, and their structures were confirmed by different structural tools. [Ru(H4dtpa) (cysteine)] Cl was separated as a model for the interaction of Ru (III) complex containing DTPA ligand with its corresponding most active amino acid and the geometrical parameters for the ternary complex and their constituent ligands are evaluated. Cytotoxicity was examined for both complexes against different cancer cell lines (HCT116, A549 and Mcf7). They were tested against a normal human lung cell line (W138). In addition, both complexes ensured cytocidal, not cytostatic effect. Several molecular tools were utilized in order to figure out the mechanism of action that two complexes followed to inhibit human colon cancer (HCT116) cell line. Gene expression analysis, cell cycle assay, genomic DNA analysis, and apoptosis were a molecular techniques that proved both Ru(H2dtpa) (H2O) and Pd (DME)Cl2 complexes were promising and effective chemotherapeutic agents against colon carcinoma in this current study. So, authors recommended more in vivo future studies should be done to make sure of complexes bioactivity and safety before they would be used commercially.

Green synthesis of nanostructured zinc oxide by Ocimum tenuiflorum extract: characterization, adsorption modeling, cytotoxic screening, and metal ions adsorption applications

Nano-ZnO was synthesized by the reduction of Zn (CH3COO)2.2H2O salt using the extract of Ocimum tenuiflorum leaves. The generated ZnO NPs were characterized by FT-IR, XRD, SEM, and EDX techniques. FT-IR results approved the characteristic peaks, the formation of ZnO bonds, and the morphology changes after the adsorption of Cd2+ and Pb2+ from solutions. The outlined data of the XRD pointed to the formation of a hexagonal wurtzite structure. SEM images showed the spherical nature of the synthesized particles with an average diameter of 19 nm. Moreover, the best conditions for the adsorption of Cd2+ and Pb2+ by ZnO NPs were evaluated and fitted to isotherm and kinetic models. Short contact time of ~ 20 min and a small sorbent dosage of 40 mg were sufficient conditions for attaining maximum Pb2+ adsorption capacity. Based on the modeling parameters, the adsorption follows pseudo-second-order kinetics where ZnO and metal ions are involved in the rate-determining step. Two important applications were thoroughly studied. The nanoparticles significantly removed Pb2+ and Cd2+ contaminants from real environmental water samples collected from different locations in Egypt. Additionally, the cytotoxic activity results provided perfect evidence for the higher efficacy of the synthesized ZnO NPs as an anticancer agent against Panc-1, PC-3, and CACO-2 cell lines with IC50 of 1.70, 3.67, and 5.70 μgml−1, respectively, compared to cisplatin (IC50 = 3.57, 5.09, and 7.75 μgml−1). Furthermore, a low cytotoxic effect was observed on the normal human lung cell line (MRC-5, IC50 = 22.40 μgml−1). The data can be used as a preliminary study for anticancer drug design after further clinical investigations.Graphical

Design, synthesis and evaluation of novel substituted fused pyrido diazepine and pyrimido piperazine derivatives: In vitro cytotoxicity study over various cancer cell lines

The current study describes, a novel combinatorial library of substituted fused pyrimido[3,2–e]pyrrolo[1,2-a]piperazine and pyrido[3,2-b]pyrrolo[1,2-d][1,4]diazepin heterocycles procured via utilization of suzuki coupling and acid amine coupling protocols. The synthesized compounds were screened for their in vitro cytotoxic activity over nine different cancer cell lines such as U-251, Panc-1, MCF-7, HepG2, DU-145, A549, 786-O, OVCAR3 and HCT-116 as well as on non-cancerous cell lines like normal human lung cell line WI38 and Mouse embryonic normal cell line NIH3T3 to evaluate safety profile of synthesized compounds. The bioassay results indicates that compound Cpd-J2 to be most potent among all the synthesized compounds towards ovary cancer cell lines (OVCAR-3) with promising activity value (EC50: 6.3 μM) relative to standard doxorubicin (EC50: 0.2 μM).

A Computational QSAR, Molecular Docking and In Vitro Cytotoxicity Study of Novel Thiouracil-Based Drugs with Anticancer Activity against Human-DNA Topoisomerase II.

Computational chemistry, molecular docking, and drug design approaches, combined with the biochemical evaluation of the antitumor activity of selected derivatives of the thiouracil-based dihydroindeno pyrido pyrimidines against topoisomerase I and II. The IC50 of other cell lines including the normal human lung cell line W138, lung cancer cell line, A549, breast cancer cell line, MCF-7, cervical cancer, HeLa, and liver cancer cell line HepG2 was evaluated using biochemical methods. The global reactivity descriptors and physicochemical parameters were computed, showing good agreement with the Lipinski and Veber’s rules of the drug criteria. The molecular docking study of the ligands with the topoisomerase protein provides the binding sites, binding energies, and deactivation constant for the inhibition pocket. Various biochemical methods were used to evaluate the IC50 of the cell lines. The QSAR model was developed for colorectal cell line HCT as a case study. Four QSAR statistical models were predicted between the IC50 of the colorectal cell line HCT to correlate the anticancer activity and the computed physicochemical and quantum chemical global reactivity descriptors. The predictive power of the models indicates a good correlation between the observed and the predicted activity.

Evaluation of Green Silver Nanoparticles Fabricated by Spirulina platensis Phycocyanin as Anticancer and Antimicrobial Agents.

Green nanotechnology has attracted attention worldwide, especially in treating cancer and drug-resistant section 6 microbes. This work aims to investigate the anticancer activity of green silver nanoparticles synthesized by Spirulina platensis phycocyanin (SPAgNPs) on two cancer cell lines: Lung cancer cell line (A-549) and breast cancer cell line (MCF-7), compared to the normal human lung cell line (A138). We also aimed to investigate the bactericidal activity against Staphylococcus aureus ATCC29737, Bacillus cereus ATCC11778, Escherichia coli ATCC8379, and Klebsiella pneumonia, as well as the fungicidal activity against Candida albicans (ATCC6019) and Aspergillus niger. The obtained SPAgNPs were spherical and crystalline with a size of 30 nm and a net charge of −26.32 mV. Furthermore, they were surrounded by active groups responsible for stability. The SPAgNPs scavenged 85% of the DPPH radical with a relative increase of approximately 30% over the extract. The proliferation of cancer cells using the MTT assay clarified that both cancer cells (A-549 and MCF-7) are regularly inhibited as they grow on different concentrations of SPAgNPs. The maximum inhibitory effect of SPAgNPs (50 ppm) reached 90.99 and 89.51% against A-549 and MCF7, respectively. Regarding antimicrobial activity, no inhibition zones occurred in bacterial or fungal strains at low concentrations of SPAgNPs and the aqueous Spirulina platensis extract. However, at high concentrations, inhibition zones, especially SPAgNPs, were more potent for all tested microorganisms than their positive controls, with particular reference to Staphylococcus aureus, since the inhibition zones were 3.2, 3.8, and 4.3 mm, and Bacillus cereus was 2.37 mm when compared to tetracycline (2.33 mm). SPAgNPs have more potent antifungal activity, especially against Aspergillus niger, compared to their positive controls. We concluded that SPAgNPs are powerful agents against oxidative stress and microbial infection.

Lab-scale Preparation of Recombinant Human Insulin-like Growth Factor-1 in Escherichia coli and its Potential Safety on Normal Human Lung Cell Line.

Insulin-like growth factor-1 (IGF-1) is structurally similar to insulin and acts as an endocrine hormone secreted by the liver. Production of recombinant human IGF-1 (rhIGF-1) in Escherichia coli (E.coli) and evaluation of its proliferation stimulatory activity. hIGF-1 gene cloned into pBSK (+) simple vector was transformed into TOP 10 chemically competent cells of E. coli. Polymerase chain reaction (PCR) was achieved using specific hIGF-1 gene primers to confirm the successful transformation. To express the rhIGF-1 in E. coli (Rosetta (DE3) pLysS); the hIGF-1 gene was cloned into the pET-15b expression vector and then the recombinant pET-15b/IGF-1 vector was transformed into a chemically prepared competent expression bacterial cells; Rosetta (DE3) pLysS. The rhIGF-1 was expressed as insoluble aggregates called inclusion bodies (IBs) using a 2 mM Isopropyl β-D-1-thiogalactopyranoside (IPTG) inducer. IBs were solubilized in a denatured form using 6 M guanidinium hydrochloride (GdmCl), followed by in vitro protein refolding using the rapid dilution method. The refolded hIGF-1 was purified using the HiTrap- ANX anion exchange column. Western blot and ELISA using rabbit polyvalent anti-hIGF- 1 were performed to confirm the protein antigenic identity. Cell proliferation activity of rhIGF-1 was testified on normal human lung cell line (WI-38). rhIGF-1 was purified from the HiTrap-ANX column at a concentration of 300 μg/ml. Western blot showed a single 7.6 kDa band obtained in the induced Rosetta (DE3) pLYsS. ELISA confirmed the molecular identity of the rhIGF-1 epitope, the concentration of purified rhIGF-1 obtained from the ELISA standard curve using rhIGF-1 reference protein as a standard was 300 μg/ml, and activity on WI-38 cells was 2604.17I U/mg. Biologically active native rhIGF-1 protein was successfully expressed. Patents related to the preparation of IGF-1 were mentioned along the text.

Inflammatory Signaling and DNA Damage Responses after Local Exposure to an Insoluble Radioactive Microparticle.

Simple SummaryA cesium-bearing microparticle (Cs-BMP) is an insoluble radioactive microparticle possessing high specific radioactivity, which was discovered after the incident at the Fukushima nuclear power plant. Due to their insoluble nature, such Cs-BMPs are assumed to adhere in the long term to normal tissue, leading to chronic local exposure. However, radiation risk due to the intake of internal exposure to radioactive cesium is conventionally estimated from the organ dose given by uniform exposure to soluble cesium. As such, it is critical to clarify the normal tissue effects posed by heterogeneous exposure to Cs-BMPs. This in vitro study reports on the relationship between the inflammatory responses and DNA damage induction during local exposure to a Cs-BMP.Cesium-bearing microparticles (Cs-BMPs) can reach the human respiratory system after inhalation, resulting in chronic local internal exposure. We previously investigated the spatial distribution of DNA damage induced in areas around a Cs-BMP; however, the biological impacts have not been fully clarified due to the limited amount of data. Here, we investigated the inflammatory signaling and DNA damage responses after local exposure to a Cs-BMP in vitro. We used two normal human lung cell lines, i.e., lung fibroblast cells (WI-38) and bronchial epithelial cells (HBEC3-KT). After 24 h exposure to a Cs-BMP, inflammation was evaluated by immunofluorescent staining for nuclear factor κB (NF-κB) p65 and cyclooxygenase 2 (COX-2). The number of DNA double-strand breaks (DSBs) was also detected by means of phospholylated histone H2AX (γ-H2AX) focus formation assay. Cs-BMP exposure significantly increased NF-κB p65 and COX-2 expressions, which were related to the number of γ-H2AX foci in the cell nuclei. Compared to the uniform (external) exposure to 137Cs γ-rays, NF-κB tended to be more activated in the cells proximal to the Cs-BMP, while both NF-κB p65 and COX-2 were significantly activated in the distal cells. Experiments with chemical inhibitors for NF-κB p65 and COX-2 suggested the involvement of such inflammatory responses both in the reduced radiosensitivity of the cells proximal to Cs-BMP and the enhanced radiosensitivity of the cells distal from Cs-BMP. The data show that local exposure to Cs-BMP leads to biological effects modified by the NF-κB pathway, suggesting that the radiation risk for Cs-BMP exposure can differ from that estimated based on conventional uniform exposure to normal tissues.

Antioxidant, antibacterial and cytotoxic effect of Cymbopogon citratus, Mentha longifolia, and Artemisia absinthium essential oils

This study aims to evaluate the benefits of some herbs such as antioxidant, antibacterial, and anti-cancer activities of Lemongrass (Cymbopogon citratus), peppermint (Mentha longifolia), and wormwood (Artemisia absinthium) essential oils (EOs). The chemical composition was identified using the GC-MS technique. The total phenolic content and the antioxidant activities were monitored by radical scavenging assay (DPPH). Furthermore, the antibacterial properties were evaluated. The possible anti-cancer activity was determined in vitro against colon (HCT116), breast (MC7) cancer, and normal human lung cell lines. The results showed that the major compounds of lemongrass EO were neral, citral,β-myrcene and camphor, while peppermint were E-Menthone, pulegone, Z-menthone, 1,8 cineole and menthol. Moreover, wormwood EO vital constituents were artemisia ketone, camphor, camphene and α-pinene. Lemongrass and wormwood EOs contain the highest total phenolic content than peppermint. Wormwood EO has the highest antioxidant activity using DPPH (IC50= 0.689%). The inhibitory effect of lemongrass, peppermint, and wormwood EOs was higher against Gram-negative bacteria. Lemongrass and wormwood EOs showed the highest anti-cancer potential against HCT116 (IC50=77.413 and IC50=297.5 µg/ml, respectively). Lemongrass and wormwood EOs effectively inhibited the HCT116 cancer cell line's growth. We recommended using these plants, which act as antioxidants, antibacterial and anti-cancer, in the future.

Recombinant Expression of Cec-B Peptide in Escherichia coli with a Significant Anticancer Effect on Hepatocellular Carcinoma.

Cecropin-B (Cec-B) is an Antimicrobial Peptide (AMP) found in insects. Recombinant production of Cec-B peptide in Escherichia coli (Rosetta™ DE3), and studying its anticancer effect on Hepatocellular Carcinoma Cell line (HCC). The Cec-B gene of Drosophila melanogaster was synthesized by PCR assembly using the Simplified Gene Synthesis (SGS) method. To express the recombinant peptide in E. coli (Rosetta™ DE3); the synthesized gene was cloned into pET-15b expression vector. The recombinant peptide was expressed as insoluble aggregates called Inclusion Bodies (IBs) using 2mM lactose inducer. IBs were solubilized in a denatured form using 8 M urea followed by in-vitro protein refolding using rapid dilution method. The refolded Cec-B was purified using cation-exchange SP-FF column. Cytotoxicity of recombinant Cec-B (rCec-B) was reported on normal human lung cell line (WI-38), and Hepatocellular carcinoma cell line (HepG2). The Cec-B gene was expressed and purified at concentration 1.212±0.1 mg/ml which represents 48.49±4% of the total proteins injected to the column (2.5±0.2 mg/ml). The safe dose of purified rCec-B on normal WI-38 cells was calculated to be 1.57 mg/ml. The half-maximal Inhibitory Concentration (IC50) of rCec-B on HepG2 cell line was calculated to be 25 μg/ml. Scanning Electron Microscope (SEM) showed that untreated and treated HepG2 cells had cell diameters from 11-12.92 μm and 14.18-21.58 μm, respectively. The results of this study revealed a successful expression of the rCec-B peptide using a pET-based expression system with a simple purification step. The purified peptide could be considered as a hopeful anticancer drug against HCC.

Antiproliferative constituents from the aerial parts of Chrysophthalmum montanum (DC.) Boiss

Three undescribed sesquiterpene lactones (1-3), 1β,4β-dihydroxy-guaia-10(14),11(13)-dien-8α,12-olide (1), 4α, 6α-dihydroxy-9β,10β-epoxy-1βH-guaia-11(13)-en-8α,12-olide (2), and 4α,9β-dihydroxy-6α-acetoxy-1βH-guaia-10(14),11(13)-dien-8α,12-olide (3), as well as a known sesquiterpene lactone, (4α,5α,8β,10α)-4,10-dihydroxy-1,11(13)-guaiadien-12,8-olide (4), along with a lignan, pinoresinol (5), a flavonoid, chrysosplenol C (6), and two triterpenes, a mixture of isomers, taraxasterol acetate and Ψ-taraxasterol acetate (7a/7b), and a mixture of isomers taraxasterol and Ψ-taraxasterol (8a/8b) were isolated from the aerial parts of Chrysophthalmum montanum (DC.) Boiss. The structures of 1-6, 7a/7b, and 8a/8b were established on the basis of spectroscopic evidence, such as MS, NMR, UV, and IR spectroscopy. All isolated compounds, except for 5 and 8a/8b, were assayed for their antiproliferative activities against three human cancer cell lines, i.e. cervical (HeLa), breast (MCF-7), and lung (A549), and a normal human lung cell line (BEAS-2B) using MTT method. Compounds 1, 4, and 6 showed significant inhibitory effects on cancer cell growth at 20 μg/mL concentration, with cell viability ranging from 53 to 64 % against MCF-7 cell line. In addition, compounds 4, and 6 exhibited cytotoxicity against HeLa cancer cell line with the viability of approximately 64 %. In conclusion, compounds 1, 4, and 6 may serve as leads for further research towards the development of anticancer agents.

Design, synthesis, and antimicrobial evaluation of 1,4-dihydroindeno[1,2-c]pyrazole tethered carbohydrazide hybrids: exploring their in silico ADMET, ergosterol inhibition and ROS inducing potential.

A series of new 1,4-dihydroindeno[1,2-c]pyrazole tethered carbohydrazide hybrids (5a-u) were designed, synthesized and evaluated for their antimicrobial activity. Compounds 5d, 5g, 5j, 5k and 5q demonstrated significant activity against the entire panel of test pathogens. Further, compounds 5d and 5g exhibited significant anti-Candida activity. These potential hybrids (5d and 5g) also exhibited promising ergosterol biosynthesis inhibition against Candida albicans, which was further validated through molecular docking studies. Furthermore, compounds 5d and 5g caused intracellular ROS accumulation in C. albicans MTCC 3017 and were non-toxic to normal human lung cell line MRC5. In silico studies revealed that they demonstrated drug likeness and an appreciable pharmacokinetic profile. Overall, the findings demonstrate that 5d and 5g may be considered as promising leads for further development of new antifungal drugs.

Some divalent metal(II) complexes of salicylaldehyde‐derived Schiff bases: Synthesis, spectroscopic characterization, antimicrobial and in vitro anticancer studies

Mononuclear transition metal(II) complexes of the type M(L)2⋅2H2O (where M = Co, Ni, Cu, Zn) have been synthesized from uninegative Schiff base ligands (HL1–HL4) designed by condensation of 4‐fluorobenzylamine with 2‐hydroxy‐1‐naphthaldehyde/3,5‐dichlorosalicylaldehyde/3,5‐dibromosalicylaldehyde/3‐bromo‐5‐chlorosalicylaldehyde. The compounds were successfully characterized using spectroscopic and physiochemical methods together with elemental analysis. Spectroscopic elucidation indicates a monobasic bidentate nature of ligands coordinated via deprotonated phenolic oxygen and azomethine nitrogen atom which suggests an octahedral geometry around the central metal ions. The complexes and ligands were screened for their in vitro antimicrobial activity against bacterial and fungal strains, the zinc(II) complexes being more active against the tested microbial strains. Further, the metal complexes were found to be more active than the uncomplexed ligands due to chelation process and, moreover, the complexes were more active against fungal strains than bacterial strains. Cytotoxic activities of all compounds were evaluated towards human alveolar adenocarcinoma epithelial cell line (A549), human breast adenocarcinoma cell line (MCF7), human prostate cancer cell line (DU145) and one normal human lung cell line (MRC‐5) using MTT colorimetric assay with doxorubicin as a standard. The zinc complexes were most active against the cancer cell lines and also found to be less toxic against MRC‐5 normal cell line than standard doxorubicin.

Induction of apoptosis in lung carcinoma cells by antiproliferative cyclic lipopeptides from marine algicolous isolate Bacillus atrophaeus strain AKLSR1

Abstract Microbes possess a vast reservoir of bioactive compounds, including cyclic lipopeptides (CLPs). In the present study, an exopolymeric biosurfactant composed of lipoheptapeptides was isolated and identified from a marine algicolous bacterial isolate, Bacillus atrophaeus strain AKLSR1, endophytic to Padina tetrastromatica. The lipopeptides were characterized by using FT-IR, NMR, GC–MS and UPLC-ESI-Q-Tof-MS/MS spectroscopic studies. Structural analysis unveiled five CLP variants that formed one major polymeric lipopeptide. These cyclic lipoheptapeptides represent a new class of surfactin family, with significant variations in the fatty acid chain length and amino acid substitutions in the peptide moiety as compared to standard lipopeptides of surfactin family. The critical micelle concentration of this biosurfactant is 0.9 mg L−1 at a surface tension of 28 mN m−1. Polymeric biosurfactant showed stable emulsification index against various organic solvents and vegetable oils. CLPs showed substantial mortality of different cancer cell lines but no toxicity towards normal human lung cell line, MRC5. Further, mechanistic studies revealed cell cycle arrest, ROS accumulation, nuclear fragmentation and cell death. Our studies indicated that CLPs induced apoptosis in lung carcinoma cell line, A549. Anti-proliferative effect of CLPs produced by marine algicolous B. atrophaeus strain AKLSR1 suggests that it could be further explored for biomedical applications.

Synthesis of novel trifluoromethyl substituted furo[2,3-b]pyridine and pyrido[3′,2′:4,5]furo[3,2-d]pyrimidine derivatives as potential anticancer agents

A series of novel trifluoromethyl substituted furo[2,3-b]pyridine and pyrido[3′,2′:4,5]furo[3,2-d] pyrimidine derivatives 3a-b, 6a-k, 9, 10a-b, 11a-c and 12a-c were prepared from 2-carbethoxy-3-amino-6-trifluoromethyl furo[2,3-b]pyridine 1 under different set of conditions. Compounds functionalized with oxadiazole 11a-c were also prepared from 2-carbohydrazide-3-amino-6-trifluoromethyl furo[2,3-b]pyridine 4. All the final products were screened for anticancer activity against four human cancer cell lines such as Neuro-2a, Hela, A549 and COLO 205 as well as normal human lung cell line, IMR-90. All the compounds showed promising anticancer activity against all the tested cell lines at <25 μM concentration except 5b, 6d, 6e and 6k. The selectivity index (SI) values have also been calculated for all the tested compounds in comparison to the normal cell line. Compounds 6g, 10a, 10b, and 11a were considered as potential leads which showed cytotoxicity with IC50 values of 10, 10.7, 11.0 and 10.5 μM, respectively. Compounds 7 and 12a were considered as highly potent exhibiting promising cytotoxicity with IC50 value of 5.8 and 3.6 μM, respectively.

Isolation and characterization of a new cytotoxic dihydrophenanthrene from Dioscorea membranacea rhizomes and its activity against five human cancer cell lines

Ethnopharmacological relevanceThe rhizomes of Dioscorea membranacea Pierre (DM) have been used as ingredients in anticancer herbal formulations in Thai traditional medicine (TTM). Thus, the aim of this study was to investigate the active constituents of DM for cytotoxic activity in order to support its TTM use. Materials and methodsA bioassay-guided isolation procedure was used to separate the cytotoxic constituents from ethanolic extract of Dioscorea membranacea rhizomes by testing against five human cancer cell lines, i.e. large cell lung carcinoma, COR-L23; liver cancer cells, HepG2; prostate cancer cells, PC3; breast cancer cells MCF-7; cervical cancer cells, Hela; and one normal human lung cell line (MRC 5) using the SRB assay. ResultsTwo known dihydrophenanthrene compounds [2,4 dimethoxy-5,6-dihydroxy-9,10-dihydrophenanthrene (1) and 5-hydroxy-2,4,6-trimethoxy-9,10-dihydrophenanthrene (2)], and a new dihydrophenanthrene compound, 5,6,2 -trihydroxy 3,4-methoxy, 9,10-dihydrophenanthrene (3) were isolated and fully characterized. 1 showed the highest cytotoxic activity against COR-L23, MCF-7 and PC3 cell lines (IC50=14.89, 17.49 and 19.04µM, respectively), and 2 showed selective cytotoxic activity against PC3 (IC50=23.54µM). The new compound 3 showed selective cytotoxic activity against only MCF-7 cells (IC50=31.41µM). Interestingly the crude extract of DM was much less toxic to the normal cell line (MRC-5) (IC50>50µg/ml) compared to the five cancer cell lines, (IC50 value ranged between 6 and 29µg/ml). ConclusionThe phytochemicals isolated from DM may serve as lead compounds for the design of new anti-cancer agents with better selective cytotoxic indices.

Genome-wide identification of cancer-related polyadenylated and non-polyadenylated RNAs in human breast and lung cell lines

Eukaryotic mRNAs consist of two forms of transcripts: poly(A)+ and poly(A)-, based on the presence or absence of poly(A) tails at the 3' end. Poly(A)+ mRNAs are mainly protein coding mRNAs, whereas the functions of poly(A)- mRNA are largely unknown. Previous studies have shown that a significant proportion of gene transcripts are poly(A)- or bimorphic (containing both poly(A)+ and poly(A)- transcripts). We compared the expression levels of poly(A)- and poly(A)+ RNA mRNAs in normal and cancer cell lines. We also investigated the potential functions of these RNA transcripts using an integrative workflow to explore poly(A)+ and poly(A)- transcriptome sequences between a normal human mammary gland cell line (HMEC) and a breast cancer cell line (MCF-7), as well as between a normal human lung cell line (NHLF) and a lung cancer cell line (A549). The data showed that normal and cancer cell lines differentially express these two forms of mRNA. Gene ontology (GO) annotation analyses hinted at the functions of these two groups of transcripts and grouped the differentially expressed genes according to the form of their transcript. The data showed that cell cycle-, apoptosis-, and cell death-related functions corresponded to most of the differentially expressed genes in these two forms of transcripts, which were also associated with the cancers. Furthermore, translational elongation and translation functions were also found for the poly(A)- protein-coding genes in cancer cell lines. We demonstrate that poly(A)- transcripts play an important role in cancer development.

Abstract B36: Transcriptional targeting of tumor cells with AAV vectors carrying diphtheria toxin A fragment

Abstract Suicide gene therapy is an attractive way of combating cancers. Diphtheria toxin (DT) has been widely studied for its application to kill cancer cells. Due to DT's extreme toxicity issue, viral vectors carrying DT are very difficult to produce since producer cells are killed before viral vectors can be produced. In this study adeno-associated viral (AAV) vectors carrying DT fragment A (DT-A) was generated at high titers in insect cells by inserting a mammalian intron to abolish the toxin expression. When these toxin carrying AAV vectors were introduced into mammalian cells, the intron was spliced out and the toxin expressed and killed the mammalian cells. Since they can direct gene expression in tumor cells but not in normal cells, tumor-specific promoters were cloned to direct the expression of the DT-A gene and AAV vectors were prepared. These vectors carrying DT-A under control of surviving or CXCR4 promoter specifically killed tumor cell lines HepG2, Hep3B, and BE (2)-M17, but not the normal human lung cell line WI38. The results in this study indicate that this novel way of targeting tumor cells with AAV vectors carrying DT-A should provide a new tool for the development of toxin-based suicide gene therapy drugs. Citation Format: Haifeng Chen. Transcriptional targeting of tumor cells with AAV vectors carrying diphtheria toxin A fragment. [abstract]. In: Proceedings of the Eleventh Annual AACR International Conference on Frontiers in Cancer Prevention Research; 2012 Oct 16-19; Anaheim, CA. Philadelphia (PA): AACR; Cancer Prev Res 2012;5(11 Suppl):Abstract nr B36.

Structural and In Vitro Biological Studies of Organotin(IV) Precursors; Selective Inhibitory Activity Against Human Breast Cancer Cells, Positive to Estrogen Receptors

Crystals of Ph3SnCl (1) were grown from a methanol/acetonitrile solution. Compounds [Ph3SnOH]n (2) and [(Ph2Sn)4Cl2O2(OH)2] (3) were crystallized from diethyl ether/methanol/acetonitrile and hot acetone/water solutions respectively, of the white precipitation, formed by adding KOH to solutions of 1 and [Ph2SnCl2] in 1 : 1 and 1 : 2 molar ratios respectively. Complex 1 was characterized by X-ray crystallography. X-ray structure determination of compounds 2 and 3 confirmed the previously reported identities. The molecular structure of 1, reported here, is a new polymorphic form of the known one for Ph3SnCl. Four independent [Ph3SnCl] molecules constitute the crystal structure of 1. The moieties are packed in two pairs in a tail-to-tail arrangement. Complexes 1–3 were evaluated for their in vitro cytotoxic activity (cell viability) against human cancer cell lines: HeLa (human cervical), MCF-7 (breast, estrogen receptor (ER) positive), MDA-MB-231 (breast, ER negative), A549 (lung), Caki-1 (kidney carcinoma), 786-O (renal adenocarcinoma), K1 (thyroid carcinoma), and the normal human lung cell line MRC-5 (normal human fetal lung fibroblast cells) versus, the normal immortalized human mammary gland epithelial cell line MTSV17 with a sulforhodamine B (SRB) assay. The results show potent cytotoxic activity of the complexes against all cell lines used, which was superior to that of cisplatin (CDDP). Compounds 1–3 showed higher activity against breast cancer cells MCF-7 (ER positive) than against of MDA-MB-231 (ER negative). These findings prompted us to search for possible interaction of these complexes with other cellular elements of fundamental importance in cell proliferation. The influence of these complexes 1–3 upon the catalytic peroxidation of linoleic acid to hydroperoxylinoleic acid by the enzyme lipoxygenase (LOX), as well as their binding affinity towards calf thymus-DNA, were kinetically and theoretically studied.

Synthesis, structural characterization and biological studies of the triphenyltin(IV) complex with 2-thiobarbituric acid

The reaction between 2-thiobarbituric acid (H 2TBA), which was treated with an equimolar amount of potassium hydroxide, in a water with triphenytin chloride in methanol, results in the formation of the {[Ph 3Sn(O-HTBA)]} n ( 1) complex. Crystals of the hydrated 1 with formula {[Ph 3Sn(O-HTBA)]·0.7(H 2O)} n were growth from methanol/acetonitrile solution, of the white precipitation, filtered off, from the reaction. The crystal structure of complex 1 has been determined by X-ray diffraction at 120 K. Complex 1 is polymeric. The geometry around the tin(IV) ions is trigonal bi-pyramidal with coordination to three C atoms from phenyl groups and one O atom from a de-protonated HTBA ligand. Complex 1 and the already known [(n-Bu) 3Sn(O-HTBA)·H 2O] ( 2) were evaluated for their in vitro cytotoxic activity (cell viability) against human cancer cell lines: HeLa (cervical), OAW-42 (ovarian), MCF-7 (breast, ER positive), MDA-MB-231 (breast, ER negative), A549 (lung), Caki-1 (renal) and additionally, the normal human lung cell line MRC-5 (normal human fetal lung fibroblast cells) and normal immortalized human mammary gland epithelial cell line MTSV17 with a Trypan Blue assay. Moreover complex 1 was evaluated for its in vitro cell growth proliferation activity against leiomyosarcoma cells (LMS), MCF-7 and MRC-5 cells with a Thiazolyl Blue Tetrazolium Bromide (MTT) assay. The type of cell death caused by complexes 1 and 2 was also evaluated by use of flow cytometry assay. The results showed that these compounds mediate a strong cytotoxic response to normal and cancer cell lines tested through apoptosis and induce cell cycle arrest in S phase of the cell cycle, suggesting DNA intercalation (direct or indirect) with the complexes. Finally, the influence of these complexes 1 and 2 upon the catalytic peroxidation of linoleic acid to hydroperoxylinoleic acid by the enzyme lipoxygenase (LOX) was kinetically and theoretically studied.