ERMP1 Facilitates The Malignant Characteristics of Colorectal Cancer Cells through Modulating PI3K/AKT/β-Catenin Pathway and Localization of GRP78

Objective: Endoplasmic reticulum-metallopeptidase 1 (ERMP1) is involved in cellular response to oxidative stress.However, its functional role in proliferation and progression of cancer cells remains unknown. The focus of this studywas to investigate the molecular-mechanisms in which ERMP1 modulates the proliferation and progression of colorectalcancer (CRC) cells under normal and environment stress conditions.Materials and Methods: In this experimental study, ERMP1 expression was evaluated using reverse transcriptionquantitativepolymerase chain reaction (RT-qPCR) in CRC cells. ERMP1 was knocked down using lentiviral transductionof ERMP1-specific shRNA into HCT116 cells. ERMP1 was also upregulated using lipofectamine transfection ofERMP1-overexpressing vector into SW48 cells. To evaluate the role of ERMP1 in the cellular and environmental stressconditions, ERMP1-downregulated cells were exposed to stressful conditions including starvation, serum free medium,and treatment with redox or chemotherapy agents for 72 hours. The expression of AKT, p-AKT, phospho-mammaliantarget of rapamycin (p-mTOR), β-catenin, p-β-catenin, E-cadherin, and Glucose-regulating protein 78 (GRP78) proteinswas evaluated by western blotting. The expression of ERMP1, CYCLIN D, and c-MYC was evaluated by RT-qPCR. Thecell surface localization of GRP78, cell cycle distribution, and apoptosis were determined by Flow cytometry.Results: ERMP1 knock-down reduced the cellular proliferation, inactivated the PI3K/AKT pathway, prompted theG1 arrest, and attenuated the free β-catenin and CYCLIN D expression. Opposite results were obtained in ERMP1-overexpressed cells. Knock-down of ERMP1 also reduced the GRP78 localization at the cell surface. Variousenvironmental stress conditions differently affected the ERMP1-downregulated cells.Conclusion: ERMP1 functioned as an oncogene in CRC cells by promoting malignant characteristics. Thephosphoinositide 3-kinases (PI3K)/AKT/β-catenin pathway and localization of GRP78 were closely related to the effectsof ERMP1. Consequently, ERMP1 might be regarded as a promising target in therapeutic strategies related to CRC

Outcomes of Implementation of Sacral Nerve Stimulation in Incontinent Patients in Shiraz

Background: Fecal incontinence is a common disorder in old age; however, it may not threaten life, but it can cause morbidity and many problems. Sacral nerve stimulation (SNS) is a minimally invasive surgical procedure performed by chronic electrical stimulation of the nerves in the sacral plexus through a lead implanted at the S3 foramen. This study aimed to evaluate the outcomes of SNS in Shiraz. Materials and Methods: Data from patients who underwent implantation of an SNS device from 2012 to 2018 were reviewed in Shiraz. Thirty patients who had incontinence were evaluated by a committee. Pre- and postoperative assessments of the severity of incontinence were performed using Wexner Incontinence Score. Statistical analysis was performed using paired t-test. Results: Twenty-seven patients proceeded to insertion in the temporary SNS, and of these, 16 were elected to have a permanent SNS. Finally, seven patients were satisfied with their treatment. There was a significant reduction in the pre- and post-SNS Wexner Incontinence Scores from a median of 15–10, respectively (P < 0.05). Conclusion: In our study, 16 patients underwent SNS protocol, and 43.7% of them showed a good response and recovered. It is recommended as a method for the treatment of fecal incontinence. Permanent SNS is effective, showing a significant improvement in fecal incontinence scores

Altered ARID1A expression in colorectal cancer

Abstract Background ARID1A has been described as a tumor suppressor gene, participating in chromatin re-modeling, epithelial-mesenchymal-transition and many other cellular and molecular processes. It has been cited as a contribute in tumorigenesis. The role of ARID1A in CRC is not yet defined. Aim To investigate the role of ARID1A methylation and CNV in its expression in CRC cell lines and to examine the relationship between ARID1A status with survival and clinicopathologic characteristics in patients with CRC. Methods We used RT-PCR to determine both CNV and expression of ARID1A from six CRC cell lines. We used MSP to evaluate methylation of ARID1A. IHC was used to assess ARID1A protein expression. We also evaluated MSI and EMAST status in 18 paired CRC and adjacent normal tissues. 5AzadC was used to assess effect of DNA demethylation on ARID1A expression. Statistical analysis was performed to establish correlations between ARID1A expression and other parameters. Results Among the 18 CRC tumors studied, 7 (38.8%) and 5 tumors (27.7%) showed no or low ARID1A expression, respectively. We observed no significant difference in ARID1A expression for overall patient survival, and no difference between clinicopathological parameters including MSI and EMAST. However, lymphatic invasion was more pronounced in the low/no ARID1A expression group when compared to moderate and high expression group (33% VS. 16.6% respectively. ARID1A promoter methylation was observed in 4/6 (66%) cell lines and correlated with ARID1A mRNA expression level ranging from very low in SW48, to more pronounced in HCT116 and HT-29/219. Treatment with the methyltransferase inhibitor 5-Azacytidine (5-aza) resulted in a 25.4-fold and 6.1-fold increase in ARID1A mRNA expression in SW48 and SW742 cells, respectively, while there was no change in SW480 and LS180 cells. No ARID1A CNV was observed in the CRC cell lines. Conclusion ARID1A expression is downregulated in CRC tissues which correlates with it being a tumor suppressor protein. This finding confirms ARID1A loss of expression in CRC development. Our in-vitro results suggest high methylation status associates with reduced ARID1A expression and contributes to CRC tumorigenesis. However, there was no significant association between ARID1A loss of expression and clinicopathological characteristics. Future in-vivo analysis is warranted to further establish ARID1A role in colorectal neoplastic transformation.http://deepblue.lib.umich.edu/bitstream/2027.42/173537/1/12885_2020_Article_6706.pd

Autophagy, Unfolded Protein Response, and Neuropilin-1 Cross-Talk in SARS-CoV-2 Infection: What Can Be Learned from Other Coronaviruses

The COVID-19 pandemic is caused by the 2019–nCoV/SARS-CoV-2 virus. This severe acute respiratory syndrome is currently a global health emergency and needs much effort to generate an urgent practical treatment to reduce COVID-19 complications and mortality in humans. Viral infection activates various cellular responses in infected cells, including cellular stress responses such as unfolded protein response (UPR) and autophagy, following the inhibition of mTOR. Both UPR and autophagy mechanisms are involved in cellular and tissue homeostasis, apoptosis, innate immunity modulation, and clearance of pathogens such as viral particles. However, during an evolutionary arms race, viruses gain the ability to subvert autophagy and UPR for their benefit. SARS-CoV-2 can enter host cells through binding to cell surface receptors, including angiotensin-converting enzyme 2 (ACE2) and neuropilin-1 (NRP1). ACE2 blockage increases autophagy through mTOR inhibition, leading to gastrointestinal complications during SARS-CoV-2 virus infection. NRP1 is also regulated by the mTOR pathway. An increased NRP1 can enhance the susceptibility of immune system dendritic cells (DCs) to SARS-CoV-2 and induce cytokine storm, which is related to high COVID-19 mortality. Therefore, signaling pathways such as mTOR, UPR, and autophagy may be potential therapeutic targets for COVID-19. Hence, extensive investigations are required to confirm these potentials. Since there is currently no specific treatment for COVID-19 infection, we sought to review and discuss the important roles of autophagy, UPR, and mTOR mechanisms in the regulation of cellular responses to coronavirus infection to help identify new antiviral modalities against SARS-CoV-2 virus