CRISPR/Cas9-mediated knockout of HO-1 decreased the proliferation and migration of T47D cells and increased cisplatin-induced apoptosis: an in vitro study.

Breast cancer is the most common type of neoplasm and the second cause of cancer-related death in women. Despite the development of novel therapeutic strategies and improved the clinical outcomes, the mortality rate for breast cancer is still high. Therefore, development of a new modality, particularly based on knocking out key genes, is under focus of investigation. Heme oxygenase-1 (HO-1) deregulation has been associated with various neoplasms-related behaviors of many types of tumor cells including breast cancer. In the current study, in order to evaluate the role of the HO-1 gene in breast cancer, we utilized the CRISPR/Cas9 technology to knock out HO-1 gene in T47D breast cancer cell line and studied its potential therapeutic effects in vitro. The cell proliferation and their sensitivity to Cisplatin were determined by CCK-8 kit. In addition, the apoptosis and the migratory potential of the cells were evaluated using Hoechst staining, and Transwell/Scratch methods, respectively. Our findings revealed that HO-1 suppression significantly reduced the proliferation ability of T47D cells (P < 0.001). Moreover, sensitivity to Cisplatin-induced toxicity increased significantly in KO-T47D cells compared to the control T47D cells. Furthermore, our findings indicated that Cisplatin-induced apoptosis increased in the KO-T47D cells. Moreover, the migratory capability of KO-T47D cells was abolished significantly (P < 0.001) as determined by Transwell migration assay. In a nutshell, our findings strongly suggest that HO-1 involved in breast cancer progression and metastasis and chemotherapy resistance. However, further comprehensive studies are required to clarify the precise role of the HO-1 gene on breast cancer cells.

Plumping up a Cushion of Human Biowaste in Regenerative Medicine: Novel Insights into a State-of-the-Art Reserve Arsenal.

Major breakthroughs and disruptive methods in disease treatment today owe their thanks to our inch by inch developing conception of the infinitive aspects of medicine since the very beginning, among which, the role of the regenerative medicine can on no account be denied, a branch of medicine dedicated to either repairing or replacing the injured or diseased cells, organs, and tissues. A novel means to accomplish such a quest is what is being called "medical biowaste", a large assortment of biological samples produced during a surgery session or as a result of physiological conditions and biological activities. The current paper accentuating several of a number of promising sources of biowaste together with their plausible applications in routine clinical practices and the confronting challenges aims at inspiring research on the existing gap between clinical and basic science to further extend our knowledge and understanding concerning the potential applications of medical biowaste.

Dimethyl fumarate prevents cytotoxicity and apoptosis mediated by oxidative stress in human adipose-derived mesenchymal stem cells.

BACKGROUND: The poor survival rate and undesirable homing of transplanted stem cells are the major challenges in stem cell therapy. Addressing the challenge would improve the therapeutic efficacy of these cells. Dimethyl fumarate (DMF) is an anti-inflammatory drug that exerts its effects through the activation of the nuclear factor erythroid 2-related factor 2 (Nrf2) pathway. Therefore, its cytoprotective effects on human adipose-derived MSCs (hASCs) against various oxidative stresses have been investigated in this study. METHODS AND RESULTS: hASCs were cultured with different concentrations of DMF to evaluate the cytotoxicity of DMF on hASCs using Cell Counting Kit-8 (CCK-8). Besides, the migration ability of the cells after DMF treatment was evaluated using the Transwell method. Furthermore, the expression of HO-1 and NQO-1 was determined using RT-PCR. The cytoprotective effects of DMF on hASCs against the oxidative stress caused by H2O2 and Ultra Violet (UV) were evaluated by assessing cell proliferation and apoptosis. Our results demonstrated that under oxidative stress conditions induced by H2O2 and UV, DMF increased the survival rate and proliferation of the cells and prevented apoptosis. Moreover, the expression of HO-1 and NQO-1 was upregulated in hASCs pretreated with DMF which confirms the activation of the Nrf2 pathway. However, DMF significantly decreased migration in hADSCs (P < 0.0001). CONCLUSION: Our findings indicate that DMF enhances the proliferation capability and viability of hASCs and prevents their apoptosis in harsh stressful microenvironments. However, the applicability of DMF as a cytoprotective factor for the augmentation of hASCs requires in-depth preclinical and clinical studies.