The effects of chitosan-loaded JQ1 nanoparticles on OVCAR-3 cell cycle and apoptosis-related gene expression.

Background and purpose: Ovarian cancer is the deadliest gynecological cancer. Bromodomain and extra terminal domain (BET) proteins play major roles in the regulation of gene expression at the epigenetic level. Jun Qi (JQ1) is a potent inhibitor of BET proteins. Regarding the short half-life and poor pharmacokinetic profile, JQ1 was loaded into newly developed nano-carriers. Chitosan nanoparticles are one of the best and potential polymers in cancer treatment. The present study aimed to build chitosan-JQl nanoparticles (Ch-J-NPs), treat OVCAR-3 cells with Ch-J-NPs, and evaluate the effects of these nanoparticles on cell cycle and apoptosis-associated genes. Experimental approach: Ch-J-NPs were synthesized and characterized. The size and morphology of Ch-J-NPs were defined by DLS and FE-SEM techniques. OVCAR-3 cells were cultured and treated with Ch-J-NPs. Then, IC50 was measured using MTT assay. The groups were defined and cells were treated with IC50 concentration of Ch-J-NPs, for 48 h. Finally, cells in different groups were assessed for the expression of genes of interest using quantitative RT-PCR. Findings/Results: IC50 values for Ch-J-NPs were 5.625 µg/mL. RT-PCR results demonstrated that the expression of genes associated with cell cycle activity (c-MYC, hTERT, CDK1, CDK4, and CDK6) was significantly decreased following treatment of cancer cells with Ch-J-NPs. Conversely, the expression of caspase-3, and caspase-9 significantly increased. BAX (pro-apoptotic) to BCL2 (anti-apoptotic) expression ratio, also increased significantly after treatment of cells with Ch-J-NPs. Conclusion and implications: Ch-J-NPs showed significant anti-cell cyclic and apoptotic effects on OVCAR-3 cells.

Upregulation of Neurotrophic Factors and Myelin Basic Protein in Schwann-like Cells by T3 Hormone Following Transdifferentiation of Human Adipose-derived Stem Cells.

Peripheral nerve regeneration is a complicated phenomenon. Thyroid hormones are known as critical regulators in the nervous system development. The Schwann cells have the regenerative potency in the peripheral nervous system. In this study, the human adipose-derived stem cells were assessed in vitro, for transdifferentiation potency into Shwann-like cells (SLCs) as a candidate source for clinical cell therapy, under the treatment of triiodothyronine (T3) hormone, and compared with the untreated cells. The cell viability rate, myelination and neurotrophic factors expression of SLCs were evaluated two weeks post- induction by MTT assay, immunocytochemistry and real-time RT-PCR techniques, respectively. The obtained results revealed a significant decrease in SLCs viability, compared to the adipose-derived stem cells (P < 0.001). Immunocytochemistry technique was applied to detect SLCs markers, such as S100ß, GFAP and myelin basic proteins (MBP) in the presence and absence of T3 treatment. The results indicated that administering T3 can significantly increase the differentiation and myelination potency of SLCs (P < 0.01). The findings of real-time RT-PCR technique indicated that the expression of Schwann cells markers, MBP, brain-derived neurotrophic factor and glial cell-derived neurotrophic factor were upregulated significantly with T3 hormone administration in comparison with the untreated cells (P < 0.05). The SLCs were able to express the neurotrophic factors and myelination related genes in the presence of T3 hormone. Furthermore, T3 administration improved myelination potency of adipose-derived stem cells, in vitro. Further in vivo experiments are necessary to confirm the advantages of using a combination of autologous SLCs and T3 hormone for peripheral nerve injury recovery.

RET Protein Expression in Colorectal Cancer; An Immunohistochemical Assessment.

BACKGROUND: RET (rearranged during transfection) is a transmembrane receptor tyrosine kinase and a receptor for the GDNF-family ligands. It plays the role of a tumor suppressor in colorectal cancer. Therefore, it is expected that RET gene becomes downregulated in colorectal cancer (CRC). In this study, we evaluated immuno-histochemical expression of RET in CRC and assessed its correlation with some of the clinicopathological features to study the prognostic value in CRC. MATERIALS AND METHODS: In total, 60 cases of colorectal cancer (CRC) from the patients who underwent surgical gastroenterology operations were randomly selected. The samples included one tumor-rich section per case and one adjacent tumor-free section as the normal control for that case. Then, immunohistochemistry (ICH) was performed for RET on all the samples and the expression of RET was analyzed. Furthermore, the correlation of RET with clinicopathological features including age, gender, location of the tumor, grade, and stage was evaluated. RESULTS: The expression of RET caused significant downregulation in cancer samples compared to the normal control ones (P = 0.002). This downregulation increased in correlation to both grade and metastasis to lymph nodes (P = 0.03 & 0.02 respectively). However, no correlation was found between the expression of RET and gender as well as location of the tumor. CONCLUSION: RET may be considered as a protein marker in CRC detection and prognosis.

Effect of Laminin on Neurotrophic Factors Expression in Schwann-Like Cells Induced from Human Adipose-Derived Stem Cells In Vitro.

The Schwann-like cells can be considered as promising in stem cell therapies, at least in experimental models. Human adipose-derived stem cells (ADSCs) are induced into Schwann-like cells (SC-like cells) and are cultured on either a plastic surface or laminin-coated plates. The findings here reveal that laminin is a critical component in extracellular matrix (ECM) of SC-like cells at in vitro. The survival rate of SC-like cells on a laminin matrix are measured through MTT assay and it is found that this rate is significantly higher than that of the cells grown on a plastic surface (P < 0.05). Schwann cell markers and the myelinogenic ability of SC-like cells at the presence versus absence of laminin are assessed through immunocytochemistry. The analysis of GFAP/S100ß and S100ß/MBP markers indicate that laminin can increase the differentiated rate and myelinogenic potential of SC-like cells. The expression levels of SCs markers, myelin basic proteins (MBP), and neurotrophic factors in two conditions are analyzed by real-time reverse transcription polymerase chain reaction (RT-PCR). The findings here demonstrated that gene expression of SCs markers, MBP, and brain-derived neurotrophic factors (BDNF) increase significantly on laminin compared to plastic surface (P < 0.01). In contrast, the nerve growth factor (NGF) expression is downregulated significantly on laminin-coated plates (P < 0.05). The obtained data suggest that production of neurotrophic factors in SC-like cell in presence of laminin can induce appropriate microenvironment for nerve repair in neurodegenerative diseases.