Synthesis and characterization of 2,3-diaminophenazine@CuhNFs and 2,3-diaminopyridine@CuhNFs: a new strategy to enhance anticancer activity

To evaluate the effect of schedule on the interaction of etoposide with paclitaxel in vitro against the A549 human lung cancer cell line and the MDA-231 and MCF-7 human breast cancer cell lines. Exposure schedules that were 24-h concurrent, 24-h sequential, and sequential 24-h with a 24-h intervening drug-free period were quantitatively evaluated by the use of the median-effect principle and the combination index. The clonogenic assay was used to assess cytotoxicity, and calculations were done with computer software. Concurrent exposures were less than additive in two of the three cell lines tested. Sequential 24-hour and sequential 24-h with an intervening 24-h drug-free period showed synergism at high effect levels in all three cell lines. Similar synergistic interactions were found when either agent was administered first. These results show a schedule-dependent cytotoxic interaction between etoposide and paclitaxel in the human lung and breast cancer cell lines evaluated, with optimal synergism occurring with sequential, but not with concurrent, treatment.

A panel of monoclonal antibodies (n = 72 including controls) directed against lung cancer antigens was screened immunohistochemically against a panel of seven human lung cancer cell lines (including small cell carcinoma, squamous cell carcinoma, adenocarcinoma and mesothelioma), six human breast cancer cell lines and one human colon cancer cell line. The majority of the antibodies (n = 42) reacted also with antigens present on breast and colon cancer cell lines. This cross reactivity especially between lung and breast cancer cell lines is not altogether unexpected since antigens common to breast and lung tissue including their neoplasms such as MUC1 antigen have been described. Our results indicate that epitopes shared by lung and breast cancers are probably more common than previously thought. The relevance for prognosis and therapy of these shared antigens, especially as disease markers in breast cancer, has to be investigated.

Reaction of 4,4-biphenyl-disulfonyl chloride with aromatic/heterocyclic sulfonamides also incorporating a free amino group, such as 4-aminobenzenesulfonamide, 4-aminoethyl-benzenesulfonamide, 6-chloro-4-aminobenzene-1,3-disulfonamide or 5-amino-1,3,4-thiadiazole-2-sulfonamide afforded bis-sulfonamides which have been tested as inhibitors of the zinc enzyme carbonic anhydrase (CA, EC 4..2.1.1). The compounds were rather modest inhibitors of isozymes CA I and XII, but were more efficient as inhibitors of the cytosolic CA II and transmembrane, tumor-associated CA IX (inhibition constants in the range of 21–129 nM gainst hCA II, and 23–79 nM against hCA IX, respectively). The new bis-sulfonamides also showed inhibition of growth of several tumor cell lines (ex vivo), with GI50 values in the range of 0.74–10.0 μg/mL against the human colon cancer cell line HCT116, the human lung cancer cell line H460 and the human breast cancer cell line MCF-7.

The insulin-like growth factors (IGFs), IGF-I and IGF-II, are potent mitogens for human lung and other epithelial cancer cell lines. Previous studies in defined medium lacking added IGF or insulin suggest that an IGF-related ligand can act as an autocrine growth factor for many cancer cell lines through action via the type I IGF receptor (IGF-R). Analysis of RNA isolated from human lung and breast cancer cell lines by reverse transcription of mRNA and polymerase chain reaction reveal that IGF-I and IGF-II mRNAs were co-expressed with IGF-R in the majority of cell lines. IGF-I mRNA was detected in 11/12 small cell lung cancer cell lines (SCLC), 13/14 nonsmall cell lung cancer (NSCLC) cell lines, and 1/2 breast cancer cell lines. IGF-II mRNA was detected in 8/10 SCLC, 11/12 NSCLC cell lines, and 2/2 breast lines. All cell lines expressed IGF-R. For analysis of IGF peptide secretion, cell lines were adapted to growth in serum/hormone-free culture medium (R0), and to avoid interference by IGF-binding proteins, secreted IGF peptides were isolated under acidic conditions and analyzed by Western blotting. Based upon measurement of the sensitivity of the anti-IGF antibodies for detection of recombinant human IGFs, IGF peptides accumulated in conditioned medium at greater than picomolar concentrations should have been readily detected. In three cell lines (two lung and one breast) secreted IGF immunoreactivity was detected as three molecular mass species of 23, 14, and 6 kDa. Isolation and NH2-terminal sequencing of each of these species definitively identified them as differentially processed forms of the IGF-II prohormone. Despite the high frequency of IGF-I gene expression detected by reverse transcription-polymerase chain reaction analysis, only one lung cancer cell line, NCI-N417d, was found that unequivocally secreted IGF-I peptide. This direct sequence determination unambiguously identifies IGF-II as the predominant IGF involved in the autocrine growth stimulation of human lung and breast epithelial tumor cell lines and supports a growing body of literature that implicates IGF-II/IGF-R autocrine loops as a common growth mechanism in epithelial carcinogenesis.

BackgroundThevetia peruviana (Pers.) K. Schum or Cascabela peruviana (L.) Lippold (commonly known as ayoyote, codo de fraile, lucky nut, or yellow oleander), native to Mexico and Central America, is a medicinal plant used traditionally to cure diseases like ulcers, scabies, hemorrhoids and dissolve tumors. The purpose of this study was to evaluate the cytotoxic, antiproliferative and apoptotic activity of methanolic extract of T. peruviana fruits on human cancer cell lines.MethodsThe cytotoxic activity of T. peruviana methanolic extract was carried out on human breast, colorectal, prostate and lung cancer cell lines and non-tumorigenic control cells (fibroblast and Vero), using the MTT assay. For proliferation and motility, clonogenic and wound-healing assays were performed. Morphological alterations were monitored by trypan blue exclusion, as well as DNA fragmentation and AO/EB double staining was performed to evaluate apoptosis. The extract was separated using flash chromatography, and the resulting fractions were evaluated on colorectal cancer cells for their cytotoxic activity. The active fractions were further analyzed through mass spectrometry.ResultsThe T. peruviana methanolic extract exhibited cytotoxic activity on four human cancer cell lines: prostate, breast, colorectal and lung, with values of IC50 1.91 ± 0.76, 5.78 ± 2.12, 6.30 ± 4.45 and 12.04 ± 3.43 μg/mL, respectively. The extract caused a significant reduction of cell motility and colony formation on all evaluated cancer cell lines. In addition, morphological examination displayed cell size reduction, membrane blebbing and detachment of cells, compared to non-treated cancer cell lines. The T. peruviana extract induced apoptotic cell death, which was confirmed by DNA fragmentation and AO/EB double staining. Fractions 4 and 5 showed the most effective cytotoxic activity and their MS analysis revealed the presence of the secondary metabolites: thevetiaflavone and cardiac glycosides.ConclusionT. peruviana extract has potential as natural anti-cancer product with critical effects in the proliferation, motility, and adhesion of human breast and colorectal cancer cells, and apoptosis induction in human prostate and lung cancer cell lines, with minimal effects on non-tumorigenic cell lines.

Chromosomal abnormalities and genomic instability in 10q22.3 region have been associated with lung cancer development, histopathological staging, and poor prognosis. Genetic mapping identified several genes including ZMIZ1, SFTPA1 and SFTPA2 in this critical region. Loss, interruption or gain of function of these genes in association with 10q22.3 deletions, translocations, or amplifications that were frequently detected in human primary lung cancer and cancer cell lines may promote oncogenesis. Human ZMIZ1, also named hZimp10, is a novel PIAS (protein inhibitor of activated STAT)-like protein that shares a zing finger domain, MIZ (MSX-interacting zinc finger), with other PIAS proteins. Fusion of ZMIZ1 to ABL1 with a t(9;10)(q34;q22.3) translocation was identified in a B-cell acute lymphoblastic leukemia patient. In this study, we performed fluorescence in situ hybridization (FISH) using genomic DNA probes covering ZMIZ1 and SFTPA gene loci at 10q22.3 to determine chromosomal aberrations involving the ZMIZ1 gene in primary lung tumor samples and lung cancer cell lines. Chromosomal abnormalities at 10q22.3 loci, including deletions, polysomy, and translocations were frequently detected in tumors and tumor-derived cell lines. We analyzed copy number variations specifically linked to ZMIZ1 gene loci by mining the published genome-wide SNP dataset from 370 NSCLC tumor samples and confirmed a significant copy number gain with average copy numbers (CN) > 5 or loss with CN < 1.5, respectively. A high frequency of loss of heterozygosity (LOH) of ZMIZ1 and ABL1 genes in lung cancer cell lines and high level amplifications of ZMIZI gene (CN > 7) in breast and prostate cancer cell lines were also detected in Sanger SNP and CGP databases, respectively. We performed Spectral karyotyping (SKY) analysis to identify chromosomal abnormalities and potential translocations involved in the 10q region in lung cancer cell lines. We found that the 10q region was translocated and fused with other chromosomes extensively in H1819 cells. This result was consistent with the high frequency 10q chromosomal abnormalities detected in human lung and other cancer cell lines in the NCI/NCBI SKY/M-FISH & CGH database. Furthermore, we developed and performed a high-throughput RACE-PCR-based gene-specific RNA sequencing analysis using next-generation parallel sequencing technology (454 Life Sciences, Inc.) to search for potential transcriptional variants and fusion partners of the ZMIZ1 gene in lung cancer cell lines which has revealed potential fusion mRNA. These results suggest that ZMIZ1 may play an important role in lung cancer development and serve as a useful biomarker for lung cancer molecular staging and prognosis. (Supported by NIH/NCI grants SPORE P50CA70907 and RO1CA116322) Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 1177.

In the present work, QSAR models for predicting the activities of ursolic acid analogs against human lung (A-549) and CNS (SF-295) cancer cell lines were developed by a forward stepwise multiple linear regression method using a leave-one-out approach. The regression coefficient (r(2)) and the cross-validation regression coefficient (rCV(2)) of the QSAR model for cytotoxic activity against the human lung cancer cell line (A-549) were 0.85 and 0.80, respectively. The QSAR study indicated that the LUMO energy, ring count, and solvent-accessible surface area were strongly correlated with anticancer activity. Similarly, the QSAR model for cytotoxic activity against the human CNS cancer cell line (SF-295) also showed a high correlation (r(2) = 0.99 and rCV(2) = 0.96), and indicated that dipole vector and solvent-accessible surface area were strongly correlated with activity. Ursolic acid analogs that were predicted to be active against these cancer cell lines by the QSAR models were semisynthesized and characterized on the basis of their (1)H and (13)C NMR spectroscopic data, and were then tested in vitro against the human lung (A-549) and CNS (SF-295) cancer cell lines. The experimental results obtained agreed well with the predicted values.

Sapindus rarak DC., Thai herbal medicine named “Ma-Gum-Dee-Kwao” in Sapindaceae family, has been widely prescribed in treatment of skin diseases and also combination with several herbal plants to treat various diseases including malignancies. Bio-based assay of water extract from S. rarak on two human breast cancer cell lines, T47D and MCF7; two human lung cancer cell lines, A549 and SK-LU1; and one colon cancer cell line, Caco-2 was investigated. We observed a significant growth inhibition only in A549 cell line with ED50 value 84.194 ± 2.597 µg/ml. Cell cycle analysis and apoptotic study were performed in A549 human lung cancer cell line and found significant increase of apoptotic cells without significant changes in cell cycle phase. Mechanism of apoptotic induction was demonstrated using Western blotting and indicated that the caspase pathway was involved in herb signal pathway. In summary, the extract-induced apoptosis of A549 cells was primarily mediated through caspase dependent pathway especially caspase 3 and 9 which suggested that S. rarak water extract might be a potential anticancer agent against human lung cancer cell line. This research might also be a hypothesis generating study in the future. Key words: Sapindus rarak, apoptosis, A549, caspase, 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) .

To investigate the effects of dalteparin sodium on the expression of vascular endothelial growth factor (VEGF), VEGF receptor (VEGFR), and hypoxia-inducible factor 1α (HIF-1α) in A549 human lung cancer (LC) cell line and a human A549-grafted nude mouse model. A549 human lung adenocarcinoma cell line was divided into control group, treated using normal saline (NS); and dalteparin sodium groups, receiving 5, 15, 50, and 150 IU/ml of dalteparin sodium, respectively. Human A549-grafted nude mouse was induced through subcutaneous (SC) injection of A549 (5 × 106/0.2 ml) into the right armpit, and randomly assigned into control group (n = 6) receiving intraperitoneal (i.p.) injection of NS, cisplatin (DDP) group (n = 6, 3 mg/kg DDP alone, i.p., for 3 days), low molecular weight heparin (LMWH) group (n = 6) receiving SC injection of 1500 IU/kg dalteparin sodium for 35 days, and DDP plus LMWH group (n = 6, 3 mg/kg DDP, i.p., for 3 days, followed by SC injection of 1500 IU/kg dalteparin sodium for 35 days). Significant difference was noted in the messenger RNA expression of VEGF, VEGFR, and HIF-1α after treating with heparin with a concentration of 15, 50, or 150 IU/ml in the A549 cell line at 24 and 48 h, respectively. In the human A549-grafted nude mouse model, a significant reduction was noted in the expression of VEGF, VEGFR, and HIF-1α in the tumor mass harvested from the mice receiving administration of dalteparin sodium plus DDP. Dalteparin sodium had the inhibitory effects on the growth of human LC A549 cells in vitro and A549 LC xenograft model, which could be enhanced when administrated together with DDP.

Irinotecan (CPT-11) is a semisynthetic camptothecin derivative with a broad spectrum of anti-tumour activity. Carboxylesterase (CE) catalyses the conversion of CPT-11 to SN-38 (7-ethyl-10-hydroxycamptothecin), the active form of CPT-11. The antiproliferative effects of CPT-11 and SN-38, CE-activity and topoisomerase I protein expression were investigated in five human small-cell lung cancer (SCLC) cell lines and four human non-small-cell lung cancer (NSCLC) cell lines. Antiproliferative activity, expressed as IC50 values, was determined using the MTT assay. CPT-11 was significantly more active in SCLC than in NSCLC cell lines (P = 0.0036), whereas no significant difference between histological types was observed with SN-38. A significant correlation (r2 = 0.52, P = 0.028) was observed between CE activity and chemosensitivity to CPT-11 but not to SN-38, and significantly higher CE activity was observed in SCLC compared with NSCLC cell lines (P = 0.025). Western blotting experiments showed topoisomerase I protein expressions within a factor of 2, and a granular nuclear staining was detectable in all cell lines by immunocytochemistry of cytospins. No correlation was observed between protein expression and sensitivity to CPT-11 or SN-38. Cellular and medium concentrations of CPT-11 and SN-38 were measured by high-performance liquid chromatography (HPLC) in one SCLC cell line with high CE activity and high sensitivity to CPT-11, and one NSCLC cell line with low sensitivity to CPT-11 and CE activity. Intracellular concentrations of CPT-11 and SN-38 were higher in the SCLC cell line, and this was associated with an increase in cellular uptake of CPT-11 compared with the medium, and an increased intracellular formation of SN-38. In conclusion, CE activity appears to be associated with higher sensitivity to CPT-11 in human lung cancer cell lines and may partly explain the difference in the in vitro sensitivity to CPT-11 between SCLC and NSCLC cells. The assessment of CE activity in clinical material of lung cancer patients undergoing treatment with CPT-11 may be warranted. However, other mechanisms may influence sensitivity to CPT-11, possibly including drug transport.

Micro-RNAs are approximately 21-25-nucleotide-long noncoding RNAs that regulate gene expression primarily at the post-transcriptional level in animals. Here, we report that micro-RNA-1 (miR-1), abundant in the cardiac and smooth muscles, is expressed in the lung and is down-regulated in human primary lung cancer tissues and cell lines. In situ hybridization demonstrated localization of miR-1 in bronchial epithelial cells. The tumor suppressor C/EBPalpha, frequently suppressed in lung cancer, reactivated miR-1 expression in the lung cancer cells. Repressed miR-1 was also activated in lung cancer cells upon treatment with a histone deacetylase inhibitor. These observations led us to examine the antitumorigenic potential of miR-1 in lung cancer cells. Expression of miR-1 in nonexpressing A549 and H1299 cells reversed their tumorigenic properties, such as growth, replication potential, motility/migration, clonogenic survival, and tumor formation in nude mice. Exogenous miR-1 significantly reduced expression of oncogenic targets, such as MET, a receptor tyrosine kinase, and Pim-1, a Ser/Thr kinase, frequently up-regulated in lung cancer. Similarly, the levels of two additional targets, FoxP1, a transcription factor with oncogeneic property, and HDAC4 that represses differentiation-promoting genes, were reduced in miR-1-expressing cells. Conversely, depletion of miR-1 facilitated N417 cell growth with concomitant elevation of these targets. Further, ectopic miR-1 induced apoptosis in A549 cells in response to the potent anticancer drug doxorubicin. Enhanced activation of caspases 3 and 7, cleavage of their substrate PARP-1, and depletion of anti-apoptotic Mcl-1 contributed to the sensitivity of miR-1-expressing cells to doxorubicin. Thus, miR-1 has potential therapeutic application against lung cancers.

Bombax ceiba is a traditional medicinal tree that is useful in the treatment of anti-tumor, anti-microbial, anti-oxidant, colds, coughs, etc. β-amyrin is a biologically active compound that was isolated from the EtOH extract of B. ceiba leaves. This work describes the anticancer activity of the leaf extract and isolated compound against human breast (MDA-MB-231 and BT-549), lung (A-549) and colon (SW-480) cancer cell lines, and the relation of the compound with anticancer activity was supported by molecular docking. The structure of the isolated compound was confirmed by the methods of 1D-NMR, 2D-NMR, and mass spectrometry data. The anticancer activity of the EtOH leaf extract and isolated compound was determined on colon, breast, and lung cancer cell lines by MTT assay. Docking studies of the isolated compound were done with the help of softwares such as Discovery Studio Visualizer, chem3D pro 12.0.2.1076, Auto Dock Tools-1.5.6, and Auto dock vina. The structure of the isolated compound was confirmed as 3β-olean-12-en-3-ol (β-amyrin). β-amyrin and the EtOH leaf extract both showed high inhibitory activities on BT-549 cancer cell line. The docking studies of the isolated compound exhibited best docking scores with PI3K and mTOR. From the best docking scores, it was clear that β-amyrin has a high probability of target of PI3Kα and mTOR. The anticancer activity of the EtOH leaf extract of Bombax ceiba and its isolated β-amyrin showed excellent anticancer activities on breast cancer cell lines and β-amyrin showed high inhibitory activities with PI3Kα and mTOR kinases by docking studies.

Introduction: A huge reservoir of bioactive compounds exists in many species of plants, only a small percentage of which have been examined and continued to be an important source of anticancer agents. Worldwide effects are ongoing to identify new anticancer compounds from plants. With the current decline in the number of new molecular entities from the pharmaceutical industry, novel anticancer agents are being sought from traditional medicines. Objective: In the present study we investigated the efficacy of methanol extracts of Pavonia odorata, for its clonogenic inhibition on Human Breast cancer (MD-MB-231), Prostate cancer (PC-3) and Lung cancer (Calu-6) cell lines. Materials and Methods: The cytotoxic effect was evaluated by MTT assay. Results: The methanol extract of P. odorata showed significant cytotoxicity against MD-MB-231 and Calu-6, when compared to PC-3cells. Conclusion: The methanol extracts of P. odorata, showed effective cytotoxic activities in a dose dependent manner. Future work will be interesting to know the chemical composition and also better understanding the mechanism of action will help in developing it as drug for therapeutic application.

BackgroundJuglans regia has been found to exhibit significant anticancer activity against various human cancer cell lines. This study was undertaken to isolate the active chemical constituent (Juglone) and to investigate its cytotoxic activity along with its various analogs against different human cancer cell lines.MethodsIsolation of juglone, a napthoquinone, from the chloroform extract of the root part of Juglans regia was executed by flash chromatography using silica gel as stationary phase. The isolated Juglone was used as starting material for the further synthesis of a novel series of triazolyl analogs using click chemistry approach to investigate their cytotoxic potential against different human cancer cell lines using 3-(4,5-Dimethylthiazol-yl)-diphenyl tetrazoliumbromide (MTT) assay.ResultsThe different extracts of Juglans regia and the isolated compound (juglone) exhibited satisfactory cytotoxic activity against a panel of eight different human cancer cell lines namely, prostate colon (Colo-205 and HCT-116), breast (T47D), prostate (PC-3 and DU-145), skin (A-431) and lung (NCI-H322 and A549). Interestingly, all the synthesised analogs displayed enhanced and selective cytotoxic activity against lung cancer cell lines only. Of the synthesized derivatives, 15a and 16a displayed the best activity with IC50 of 4.72 and 4.67 μM against A549 cells. Both these derivatives exhibited superior potency to BEZ-235 against both the lung cancer cell lines. So far as the structural aspects are concerned, electron withdrawing substituents at the ortho position of R moiety of the triazolyl analogs seem to be essential for attaining better activity.ConclusionThe present study demonstrates the selective and enhanced cytotoxic activity of the triazolyl analogs of juglone against NCI-H322 and A549 human lung cancer cell lines. Some derivatives exhibited superior potency to BEZ-235, a commercially available anticancer agent.

In the present study, an eco-friendly approach is adapted for the synthesis of reduced graphene oxide (rGO's) by a simple hydrothermal reaction using two plant extracts namely Acalypha indica and Raphanus sativus. After the hydrothermal reaction, GO turns into a black color from brown color, which indicates the successful reduction of graphene oxide. Further, various characterization techniques such as UV-Vis spectroscopy, Raman spectroscopy, Fourier transform infrared spectroscopy (FT-IR), and X-ray diffraction is used to confirm the physicochemical properties of synthesized rGO's. Raman analysis confirms the reduction of GO by noticing an increase in the ID/IG ratio significantly. Field emission scanning electron microscopy and transmission electron microscopy clearly show the morphology and crystalline nature of rGO's. FT-IR spectrum confirms that the bioactive molecules of the plant extract (i.e. polyphenols, flavonoids, terpenoids, etc.) playing a key role in the elimination of oxygen groups from the GO surface. Further, the synthesized rGO's are tested for their potential against human lung and breast cancer cell lines. A significant cancer cell inhibition activity is obtained even in the less concentration of rGO's with IC50 values for lung cancer cell lines are 38.46µg/mL and 26.69µg/mL for AIrGO and RSrGO, respectively. Similarly, IC50 values for breast cancer cell lines are 35.97µg/mL and 33.22µg/mL for AIrGO and RSrGO, respectively.