Morphomolecular identification, metabolic profile, anticancer, and antioxidant capacities of Penicillium sp. NRC F1 and Penicillium sp. NRC F16 isolated from Egyptian remote cave

Searching for remote locations to screen for microorganisms, identify their metabolites, and investigate their bioactivities against lethal diseases such as cancer is of critical importance. In the current study, two fungal strains where isolated from a remote cave in Asyut governorate, Egypt. These isolates were morphologically and molecularly identified through sequencing their ITS region as Penicillium sp. NRC F1, and Penicillium sp. NRC F16. Investigating the metabolic profiles of the silylated ethyl acetate extracts of these fungi through conducting GC-Ms analysis revealed presence of 114 compounds belonging to different chemical classes. On the other hand, studying the in vitro bioactivity of both extracts showed moderate antioxidant activities. Penicillium sp. NRC F1 extract exhibited higher DPPH scavenging activity (74.41 ± 0.59%) at concentration of 200 µg/ml, in comparison with that exerted by Penicillium sp. NRC F16 extract at the same concentration (65.58 ± 1.55%). Moreover, studying the cytotoxicity of extracts against human colon cancer (HCT116), and human breast cancer (MCF7) cell lines revealed that cytotoxicity of both extracts was dose dependent. Promising cytotoxic effect was achieved against human colon cancer HCT116 using 200 µg/ml of Penicillium sp. NRC F1 extract (95.72 ± 1.13 % cytotoxicity), while Penicillium sp. NRC F16 ethyl acetate extract caused a cytotoxicity of 95.43 ± 1.4 %. Similarly, investigating the in vitro cytotoxicity of the extracts against human breast cancer MCF7 cell line resulted in observing promising activity of Penicillium sp. NRC F1 and Penicillium sp. NRC F16 extracts, and they exhibited 97.29 ± 0.61 %; and 97.08 ± 1.07 % cytotoxicity, respectively. Results from this study nominate those strains as promising isolates and encourage for conducting further in vivo investigations to evaluate their potency.

1,3,4-Oxadiazole N-Mannich Bases: Synthesis, Antimicrobial, and Anti-Proliferative Activities

The reaction of 5-(3,4-dimethoxyphenyl)-1,3,4-oxadiazole-2(3H)-thione 3 with formaldehyde solution and primary aromatic amines or 1-substituted piperazines, in ethanol at room temperature yielded the corresponding N-Mannich bases 3-arylaminomethyl-5-(3,4-dimethoxyphenyl)-1,3,4-oxadiazole-2(3H)-thiones 4a–l or 3-[(4-substituted piperazin-1-yl)methyl]-5-(3,4-dimethoxyphenyl)-1,3,4-oxadiazole-2(3H)-thiones 5a–d, respectively. The in vitro inhibitory activity of compounds 4a–l and 5a–d was assessed against pathogenic Gram-positive, Gram-negative bacteria, and the yeast-like pathogenic fungus Candida albicans. The piperazinomethyl derivatives 5c and 5d displayed broad-spectrum antibacterial activities the minimal inhibitory concentration (MIC) 0.5–8 μg/mL) and compounds 4j, 4l, 5a, and 5b showed potent activity against the tested Gram-positive bacteria. In addition, the anti-proliferative activity of the compounds was evaluated against prostate cancer (PC3), human colorectal cancer (HCT-116), human hepatocellular carcinoma (HePG-2), human epithelioid carcinoma (HeLa), and human breast cancer (MCF7) cell lines. The optimum anti-proliferative activity was attained by compounds 4l, 5a, 5c, and 5d.

MicroRNA-126-5p Inhibits the Migration of Breast Cancer Cells by Directly Targeting CNOT7

Background: To assess the effect of microRNA-126-5p (miR-126-5p) on the migration of the breast cancer MCF7 cell line. Methods: GSE143564 was downloaded from the Gene Expression Omnibus (GEO; http://www.ncbi.nlm.nih.gov/geo ) to identify the differentially expressed miRNAs between breast cancer and adjacent tissues. Quantitative reverse transcription PCR (RT-qPCR) was used to assess miR-126-5p levels in the normal 184A1 breast cell line and the breast cancer MCF7 cell line. The MCF7 cell line was then transfected with miR-126-5p mimics or corresponding negative control (NC-mimic). The proliferation and migration abilities of the MCF7 cell line were measured by methyl thiazolyl tetrazolium (MTT), Transwell and scratch healing assays. CCR4-NOT transcription complex and subunit 7 (CNOT7) expression levels in the NC-mimic and miR-126-5p mimic groups were measured by Western blot analysis. Bioinformatic analysis and a dual-luciferase reporter assay were performed to identify the miR-126-5p target gene. Results: One hundred forty-eight differentially expressed miRNAs (downregulated = 55, upregulated = 93) were identified. MiR-126-5p expression in the MCF7 cell line was significantly downregulated relative to that of 184A1 cell line (P < 0.05). Compared with that observed in the control and NC-mimic groups, cell proliferation in the miR-126-5p mimic group was significantly decreased at 48 and 72 h posttransfection (P < 0.05). In addition, the scratch healing rate and number of membrane-piercing cells in the miR-126-5p overexpression group were lower than those detected in the control and NC groups (P < 0.05). Furthermore, miR-126-5p could reduce the luciferase activity for the wild-type CNOT7 gene 3’-untranslated region (UTR) reporter (P < 0.05) but had no effect on the mutant 3’UTR reporter (P > 0.05). Compared with that observed in the NC and control groups, the levels of CNOT7 in the miR-126-5p overexpression group decreased (P < 0.05). Conclusion: Upregulation of miR-126-5p can inhibit the migration of the breast cancer MCF7 cell line, which may involve its direct targeting of the 3’UTR of CNOT7.

SYNTHESIS AND BIOLOGICAL EVALUATION OF 2-PHENYLPYRIDO[2,3-D] PYRIMIDINE DERIVATIVES AS CYCLIN-DEPENDENT KINASE (CDK) INHIBITORS

We report a novel scaffold of 2-phenylpyrido[2,3-d]pyrimidine derivatives designed as structural analogues of dinaciclib. Sixteen derivatives were synthesised and evaluated for their CDK2/5 inhibition activity. Compounds 4-(2-(3-methoxybenzylidene)hydrazineyl)-2-phenylpyrido[2,3-d]pyrimidine (7i) and 4-(2-(3-nitrobenzylidene)hydrazineyl)-2-phenylpyrido[2,3-d]pyrimidine (7n) show promising IC50 and kinase selectivity. These compounds also show moderate anti-proliferative activity in the colon cancer HCT116 and breast cancer MCF7 cell lines. In molecular docking studies with CDK2, compounds 7i and 7nshow binding similar to dinaciclib.

CYTOTOXIC EFFECTS OF ETHYL ACETATE FRACTIONS FROM SECONDARY METABOLITES OF STREPTOMYCES SP. GMY01 ON HUMAN BREAST CANCER MCF7 CELL LINES

Objective: This research aimed to fractionate the ethyl acetate extract from secondary metabolites of Streptomyces sp. GMY01 and to identify which fraction contains cytotoxic active compounds against human breast cancer MCF7 cell lines.Methods: Secondary metabolites were obtained from fermentation of Streptomyces Sp. GMY01 for 15 d. The supernatant containing these secondary metabolites was extracted through partition using ethyl acetate as the solvent. Fractionation of the ethyl acetate extract was conducted via column chromatography using silica gel as the solid phase while the gradient mobile phase consisted of n-hexane, ethyl acetate, and methanol. The cytotoxicity of each fraction was calculated using MTT-assay.Results: The ethyl acetate extract could be separated into 9 fractions using column chromatography. The cytotoxic effect of each fraction differed from each other. The smallest IC50 value was obtained from fraction 4. Further investigation should be conducted to discover the active anticancer compound. The active compound with cytotoxic effect was found in fraction 4 because of the highest IC50 value.Conclusion: This fraction is potential to be investigated more deeply as anticancer, especially for breast cancer.

Antiproliferative Activity of Xanthones Isolated fromArtocarpus obtusus

An investigation of the chemical constituents inArtocarpus obtususspecies led to the isolation of three new xanthones, pyranocycloartobiloxanthone A (1), dihydroartoindonesianin C (2), and pyranocycloartobiloxanthone B (3). The compounds were subjected to antiproliferative assay against human promyelocytic leukemia (HL60), human chronic myeloid leukemia (K562), and human estrogen receptor (ER+) positive breast cancer (MCF7) cell lines. Pyranocycloartobiloxanthone A (1) consistently showed strong cytotoxic activity against the three cell lines compared to the other two with IC50values of 0.5, 2.0 and 5.0 μg/mL, respectively. Compound (1) was also observed to exert antiproliferative activity and apoptotic promoter towards HL60 and MCF7 cell lines at respective IC50values. The compound (1) was not toxic towards normal cell lines human nontumorigenic breast cell line (MCF10A) and human peripheral blood mononuclear cells (PBMCs) with IC50values of more than 30 μg/mL.