METHODS: Therefore, the present study aimed to investigate the messenger ribonucleic acid (mRNA) expression of BRS3 in human liver THLE-2 cells post-BPA treatment by real-time polymerase chain reaction. The effects of BPA on the levels of pro-inflammatory proteins, interleukin 6 (IL6) and CC motif chemokine ligand 2 (CCL2), in conditioned media of BPA-treated THLE-2 cells and deoxyribonucleic acid (DNA) synthesis in replicating BPA-treated THLE-2 cells during the cell cycle were also examined by enzyme-linked immunosorbent assay (ELISA) and flow cytometry, respectively.
RESULTS: The study found that the mRNA expression of BRS3 was increased in THLE-2 cells treated with BPA. The study also showed that the expression levels of IL6 and CCL2 reached an optimum level in the conditioned media of BPA-treated THLE-2 cells after 48 h of treatment. Subsequently, the DNA synthesis analysis showed that bromodeoxyuridine/propidium iodide (BrdU/PI) stained positive cells were decreased in BPA-treated THLE-2 cells at 72 h of treatment.
CONCLUSION: The study demonstrates that BRS3 expression induced by BPA is likely associated with reduced cell proliferation by inhibiting DNA synthesis and inducing cellular inflammation in liver cells.
METHODS: The proliferative effect of the identified protein on MCF-7 cells that interacted non-adhesively with BMSCs pre-treated with pioglitazone and/or rosiglitazone was evaluated using cell culture inserts and conditioned media. The mRNA expression of proliferation and lipid accumulation markers was also evaluated in the interacted MCF-7 cells by reverse transcription-quantitative PCR. Finally, the correlation between the identified protein and fibroblast growth factor 4 (FGF-4) protein in the conditioned media of the pre-treated BMSCs was evaluated by ELISA.
RESULTS: The present study identified vimentin as the specific protein among the complex intracellular proteins that likely plays a role in MCF-7 cell proliferation when the breast cancer cells interacted non-adhesively with BMSCs pre-treated with a combination of pioglitazone and rosiglitazone. The inhibition of this protein promoted the proliferation of MCF-7 cells when the breast cancer cells interacted with pre-treated BMSCs. Gene expression analysis indicated that pre-treatment of BMSCs with a combination of pioglitazone and rosiglitazone decreased the mRNA expression of Ki67 and proliferating cell nuclear antigen in MCF-7 cells. The pre-treatment did not induce mRNA expression of PPARγ, which is a sign of lipid accumulation. The level of vimentin protein was also associated with the FGF-4 protein expression level in the conditioned media of the pre-treated BMSCs. Bioinformatics analysis revealed that vimentin regulated the expression of FGF-4 through its interaction with SRY-box 2 and POU class 5 homeobox 1.
CONCLUSIONS: The present study identified a novel intracellular protein that may represent the promising target in pre-treated BMSCs to decrease the proliferation of breast cancer MCF-7 cells for human health and wellness.
METHODS: Faecal and gut microbiota of Columbia livia were isolated, identified and conditioned media were prepared containing metabolites. Growth inhibition, lactate dehydrogenase cytotoxicity and cell survival assays were accomplished against cervical cancer cells. Next, liquid-chromatography mass spectrometry was conducted to elucidate the molecules present.
RESULTS: A plethora of bacteria from faecal matter and gastrointestinal tract were isolated. Selected conditioned media exhibited potent anticancer effects and displayed cytotoxicity to cervical cancer cells at IC50 concentration of 10.65 and 15.19 µg/ml. Moreover, cells treated with conditioned media exhibited morphological changes, including cell shrinking and rounding; indicative of apoptosis, when compared to untreated cells. A total of 111 and 71 molecules were revealed from these gut and faecal metabolites. The identity of 60 molecules were revealed including, dihydroxymelphalan. Nonetheless, 122 molecules remain unidentified and are the subject of future studies.
CONCLUSION: These findings suggest that gut bacteria of Columbia livia possess molecules, which may have anticancer activities. Further in silico testing and/or high throughput screening will determine potential anticancer properties of these molecules.
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SIGNIFICANCE AND IMPACT OF THE STUDY: The bacteria isolated from unusual dwellings such as the cockroaches' gut are a useful source of antibacterial and antiamoebal molecules. These are remarkable findings that will open several avenues in our search for novel antimicrobials from unique sources. Furthermore studies will lead to the identification of molecules to develop future antibacterials from insects.