Antitumor Activity of Ethanolic Extract of Dendrobium formosum in T-Cell Lymphoma: An In Vitro and In Vivo Study

Dendrobium, a genus of orchid, was found to possess useful therapeutic activities like anticancer, hypoglycaemic, antimicrobial, immunomodulatory, hepatoprotective, antioxidant, and neuroprotective activities. The study was aimed to evaluate the anticancer property of the ethanolic extract of Dendrobium formosum on Dalton’s lymphoma. In vitro cytotoxicity was determined by MTT assay, apoptosis was determined by fluorescence microscopy, and cell cycle progression was analysed using flow cytometry; in vivo antitumor activity was performed in Dalton’s lymphoma bearing mice. The IC50 value of ethanolic extract was obtained at 350 μg/mL in Dalton’s lymphoma cells. Fluorescence microscopy analysis showed significant increase in apoptotic cell death in dose- and time-dependent manner which was further confirmed through the resulting DNA fragmentation. Further, flow cytometry analysis showed that the ethanolic extract arrests the cells in G2/M phase of the cell cycle. The in vivo anticancer activity study illustrates significant increase in the survival time of Dalton’s lymphoma bearing mice on treatment with ethanolic extract when compared to control. These results substantiate the antitumor properties of ethanolic extract of Dendrobium formosum and suggest an alternative in treatment of cancer. Further studies are required regarding the isolation and characterization of bioactive components along with the analysis of molecular mechanism involved

CoCl2 simulated hypoxia induce cell proliferation and alter the expression pattern of hypoxia associated genes involved in angiogenesis and apoptosis

Abstract Background/aims Hypoxia microenvironment plays a crucial role during tumor progression and it tends to exhibit poor prognosis and make resistant to various conventional therapies. HIF-1α acts as an important transcriptional regulator directly or indirectly associated with genes involved in cell proliferation, angiogenesis, apoptosis and energy metabolism during tumor progression in hypoxic microenvironment. This study was aimed to investigate the expression pattern of the hypoxia associated genes and their association during breast cancer progression under hypoxic microenvironment in breast cancer cells. Methods Cell proliferation in MCF-7 and MDA-MB-231 cell lines treated with different concentration of CoCl2 was analyzed by MTT assay. Flow cytometry was performed to check cell cycle distribution, whereas cell morphology was examined by phase contrast microscopy in both the cells during hypoxia induction. Expression of hypoxia associated genes HIF-1α, VEGF, p53 and BAX were determined by semiquantitative RT-PCR and real-time PCR. Western blotting was performed to detect the expression at protein level. Results Our study revealed that cell proliferation in CoCl2 treated breast cancer cells were concentration dependent and varies with different cell types, further increase in CoCl2 concentration leads to apoptotic cell death. Further, accumulation of p53 protein in response to hypoxia as compare to normoxia showed that induction of p53 in breast cancer cells is HIF-1α dependent. HIF-1α dependent BAX expression during hypoxia revealed that after certain extent of hypoxia induction, over expression of BAX conquers the effect of anti-apoptotic proteins and ultimately leads to apoptosis in breast cancer cells. Conclusion In conclusion our results clearly indicate that CoCl2 simulated hypoxia induce the accumulation of HIF-1α protein and alter the expression of hypoxia associated genes involved in angiogenesis and apoptosis

Isolation and Characterization of “Terrein” an Antimicrobial and Antitumor Compound from Endophytic Fungus Aspergillus terreus (JAS-2) Associated from Achyranthus aspera Varanasi, India

The present study aimed at characterizing biological potentials of endophyte Aspergillus terreus JAS-2 isolated from Achyranthus aspera. Crude extracted from endophytic fungus JAS-2 was purified and chemically characterized by chromatographic and spectroscopic studies respectively. Spectral assignment of NMR (nuclear magnetic resonance) data, 1H proton and 13C carbon analysis along with FTIR data elucidated the structure of compound as 4,5-Dihydroxy-3-(1-propenyl)-2-cyclopenten-1-one. After purification and identification a set of experiment was conducted to explore efficacy of compound. Results revealed that on accessing the antifungal activity of compound, growth diameter of tested phytopathogenic fungi was reduced to 50% at higher concentration taken (10 μgμl−1). Compound exhibited in-vitro bacterial cell inhibition at 20 μgml−1 concentration along with moderate antioxidant behavior. Evaluation of anticancer activity against human lung cancer cell line (A-549) exhibited its IC50 value to be 121.9 ± 4.821 μgml−1. Further cell cycle phase distribution were analyzed on the basis of DNA content and evaluated by FACS (Fluorescence Activated Cell Sorting) and it was revealed that at 150 μgml−1 of compound maximum cells were found in sub G1 phase which represents apoptotic dead cells. Terrein (4, 5-Dihydroxy-3-(1-propenyl)-2-cyclopenten-1-one) a multi-potential was isolated from endophytic fungus JAS-2, from well recognized medicinal herb A. aspera. To best of our knowledge, this is the first report of “Terrein” from endophytic derived fungus. This compound had also exhibited anticancer and antifungal activity against human lung cancer cell line A-549 and Bipolaris sorokiniana respectively which is causal organism of many plants disease. Hence endophytes are serving as alternative sources of drug molecules

Interaction of ferrocene appended Ru(II), Rh(III) and Ir(III) dipyrrinato complexes with DNA/protein, molecular docking and antitumor activity

Efficacy of the ferrocene appended piano-stool dipyrrinato complexes [(η6-C6H6)RuCl(fcdpm)](1), [(η6C10H14)RuCl(fcdpm)](2), [(η6-C12H18)RuCl(fcdpm)](3) [(η5-C5Me5)RhCl(fcdpm)](4) and [(η5-C5Me5IrCl(fcdpm)] (5) [fcdpm = 5-ferrocenyldipyrromethene] toward anticancer activity have been described. Binding of the complexes with calf thymus DNA (CT-DNA) and BSA (bovine serum albumin) have been thoroughly investigated by UV–Vis and fluorescence spectroscopy. Binding constants for 1–5 (range, 104–105 M-1) validated their efficient binding with CT-DNA. Molecular docking studies revealed interaction through minor groove of the DNA, on the other hand these also interact through hydrophobic residues of the protein, particularly cavity in the subdomain IIA. In vitro anticancer activity have been scrutinized by MTT assay, acridine orange/ethidium bromide (AO/EtBr) fluorescence staining, and DNA ladder (fragmentation) assay against Dalton's Lymphoma (DL) cells. Present study revealed that rhodium complex (4) is more effective relative to ruthenium (1–3) and iridium (5) complexes

Potential apoptosis inducing agents based on a new benzimidazole schiff base ligand and its dicopper(II) complex

Synthesis, characterization and antiproliferative activity of a new benzimidazole based Schiff base 2-(1-methyl-1-H-benzimidazol-2-yl)phenyl)imino)methyl)phenol (HL) and dicopper(II) complex [{Cu(L)NO3}<SUB>2</SUB>] (CuL) containing L<SUP>−</SUP> has been described. Both HL and CuL have been meticulously characterized by satisfactory elemental analyses, FT-IR, NMR, ESI-MS, electronic absorption and emission spectroscopy, and their structures unambiguously determined by X-ray single crystal analyses. Titration studies (absorption and emission) revealed interaction of the ligand and its dicopper(II) complex with DNA/BSA and stronger affinity of the CuL relative to HL. Binding of the HL and CuL with DNA/BSA have been validated by in silico studies and their cytotoxic effect on human breast cancer cell lines (MCF-7) by MTT assay. IC<SUB>50</SUB> values (458 μM and 22 μM for HL, CuL) clearly suggested substantial cytotoxicity of the complex CuL toward MCF-7 compared to the ligand HL. Greater antiproliferative efficacy of the CuL in contrast to HL has been evidenced by fluorescence activated cell sorting (FACS) and AO/EB fluorescence staining. The possible mode of the apoptotic pathway for CuL has further been affirmed by reactive oxygen species (ROS) generation studies

DNA binding and anti-cancer activity of redox-active heteroleptic piano-stool Ru(II), Rh(III), and Ir(III) complexes containing 4-(2-methoxypyridyl)phenyldipyrromethene

The synthesis of four novel heteroleptic dipyrrinato complexes [(η<SUP>6</SUP>-arene)RuCl(2-pcdpm)] (η<SUP>6</SUP>-arene = C<SUB>6</SUB>H<SUB>6</SUB>, 1; C<SUB>10</SUB>H<SUB>14</SUB>, 2) and [(η<SUP>5</SUP>-C<SUB>5</SUB>Me<SUB>5</SUB>)MCl(2-pcdpm)] (M = Rh, 3; Ir, 4) containing a new chelating ligand 4-(2-methoxypyridyl)-phenyldipyrromethene (2-pcdpm) have been described. The complexes 1–4 have been fully characterized by various physicochemical techniques, namely, elemental analyses, spectral (ESI-MS, IR, <SUP>1</SUP>H, <SUP>13</SUP>C NMR, UV/vis) and electrochemical studies (cyclic voltammetry (CV) and differential pulse voltammetry (DPV)). Structures of 3 and 4 have been determined crystallographically. In vitro antiproliferative and cytotoxic activity of these complexes has been evaluated by trypan blue exclusion assay, cell morphology, apoptosis, acridine orange/ethidium bromide (AO/EtBr) fluorescence staining, and DNA fragmentation assay in Dalton lymphoma (DL) cell lines. Interaction of 1–4 with calf thymus DNA (CT DNA) has also been supported by absorption titration and electrochemical studies. Our results suggest that in vitro antitumor activity of 1–4 lies in the order 2 > 1 > 4 > 3

Synthesis, DNA binding, cellular DNA lesion and cytotoxicity of a series of new benzimidazole-based Schiff base copper(II) complexes

A series of new benzimidazole containing compounds 2-((1-R-1-H-benzimidazol-2-yl)phenyl-imino)naphthol HL1–3 (R = methyl, ethyl or propyl, respectively) have been synthesized by Schiff base condensation of 2-(1-R-1-H-benzo[d]imidazol-2-yl)aniline and 2-hydroxy-1-naphthaldehyde. The reactions of HL1–3 with Cu(NO3)2·2.5H2O led to the corresponding copper(II) complexes [Cu(L)(NO3)] 1–3. All the compounds were characterized by conventional analytical techniques and, for 1 and 3, also by single-crystal X-ray analysis. The interactions of complexes 1–3 with calf thymus DNA were studied by absorption and fluorescence spectroscopic techniques and the calculated binding constants (Kb) are in the range of 3.5 × 105 M−1–3.2 × 105 M−1. Complexes 1–3 effectively bind DNA through an intercalative mode, as proved by molecular docking studies. The binding affinity of the complexes decreases with the size increase of the N-alkyl substituent, in the order of 1 > 2 > 3, which is also in accord with the calculated LUMOcomplex energies. They show substantial in vitro cytotoxic effect against human lung (A-549), breast (MDA-MB-231) and cervical (HeLa) cancer cell lines. Complex 1 exhibits a significant inhibitory effect on the proliferation of the A-549 cancer cells. The antiproliferative efficacy of 1 has also been analysed by a DNA fragmentation assay, fluorescence activated cell sorting (FACS) and nuclear morphology using a fluorescence microscope. The possible mode for the apoptosis pathway of 1 has also been evaluated by a reactive oxygen species (ROS) generation study

Intracellular detection of Cu2+ and S2− ions through a quinazoline functionalized benzimidazole-based new fluorogenic differential chemosensor

A new quinazoline functionalized benzimidazole-based fluorogenic chemosensor H3L is synthesized and fully characterized by conventional techniques including single crystal X-ray analysis. It acts as a highly selective colorimetric and fluorescence sensor for Cu2+ ions in DMF/0.02 M HEPES (1 : 1, v/v, pH = 7.4) medium. Reaction of H3L with CuCl2 forms a mononuclear copper(II) [Cu(Cl)(H2L)(H2O)] (H2L–Cu2+) complex which is characterized by conventional techniques and quantum chemical calculations. Electronic absorption and fluorescence titration studies of H3L with different metal cations show a distinctive recognition only towards Cu2+ ions even in the presence of other commonly coexisting ions such as Li+, Na+, K+, Mg2+, Ca2+, Fe2+, Fe3+, Mn2+, Co2+, Ni2+, Zn2+, Cd2+ and Hg2+. Moreover, H2L–Cu2+ acts as a metal based highly selective and sensitive chemosensor for S2− ions even in the presence of other commonly coexisting anions such as F−, Cl−, Br−, I−, SO42−, SCN−, AcO−, H2PO4−, PO43−, NO3−, ClO4−, NO2−, HSO4−, HSO42−, S2O32−, S2O82−, CN−, CO32− and HCO3− in DMF/0.02 M HEPES (1 : 1, v/v, pH = 7.4) medium. Quantification analysis indicates that these receptors, H3L and H2L–Cu2+, can detect the presence of Cu2+ and S2− ions at very low concentrations of 1.6 × 10−9 M and 5.2 × 10−6 M, respectively. The propensity of H3L as a bio-imaging fluorescent probe for detection of Cu2+ ions and sequential detection of S2− ions by H2L–Cu2+ in Dalton lymphoma (DL) cancer cells is also shown