Overexpression of the non-coding SOX2OT variants 4 and 7 in lung tumors suggests an oncogenic role in lung cancer

Despite the advances in cancer therapy, lung cancer still remains the most leading cause of cancer death worldwide. The long non-coding RNAs (lncRNAs) are recently introduced as novel regulators of human cancers. SOX2 overlapping transcript (SOX2OT) is a cancer-associated lncRNA gene that encodes different alternatively spliced transcripts. Here, we investigated the alterations in the preferential expression of different SOX2OTs in twenty non-small cell lung cancer (NSCLC) patients by real-time quantitative reverse transcription PCR (qRT-PCR) method. We observed preferential expression of SOX2OT4 and SOX2OT7 in lung tumor tissues. The quantitative gene expression analysis revealed that >30 % of NSCLC tumors express SOX2OT4 (mean = 7.6 times) and SOX2OT7 (mean = 5.9 times) more than normal tissues, with higher expression in squamous cell carcinoma. Further, we observed overexpression of pluripotency-associated transcription factor, SOX2 in 47 % of our samples concordant with SOX2OT (R = 0.62, P value <0.05). Overexpression of OCT4A gene was also observed in 36.8 % of tumor tissues. Then, we investigated the effects of SOX2OT suppression in lung adenocarcinoma cell line, by means of RNAi. Cell characteristics of colony formation, apoptosis, 2-D mobility, and cell cycle progression were measured in control and treated A549 cells. The SOX2OT knockdown significantly reduced the colony formation ability of cancer cells; however, no alterations in the rate of apoptosis were detected. On the other hand, SOX2OT-suppressed cells had elevated accumulation in G2/M phase of cell cycle and exhibited limited mobility. Altogether, our findings support a potential oncogenic role for SOX2OT in non-small cell lung cancer tumor genesis and SOX2OT seems a promising therapeutic candidate for NSCLC. © 2016, International Society of Oncology and BioMarkers (ISOBM)

Identification of new SOX2OT transcript variants highly expressed in human cancer cell lines and down regulated in stem cell differentiation

Long non-coding RNAs are manifested as a new paradigm of molecular effectors in a wide range of human diseases. Human SOX2 overlapping transcript (SOX2OT) gene can generate six lncRNA transcript variants which are functionally assumed to be correlated with cellular differentiation and carcinogenesis. However, the circumstances determining expressional and functional differences between SOX2OT transcript variants remain to be explored. Here, we studied the expression of all SOX2OT transcript variants specifically in five human cancer cell lines by real-time RT-PCR. Changes of the new SOX2OT transcript variants expression were measured during the NT2 teratocarcinoma cell line neuronal-like differentiation and were compared to pluripotency regulators, SOX2 and OCT4A gene expressions. Surprisingly, we identified two new SOX2OT transcripts, named SOX2OT-7, SOX2OT-8 which lack exon 8. We discovered that beside active proximal and distal SOX2OT promoters, different cancer cell lines express high levels of some SOX2OT transcript variants differentially by alternative splicing. Significantly, both SOX2OT-7 and SOX2OT-8 are highly expressed in human cancer cell lines coinciding with SOX2, one of the pluripotency regulators. Our results revealed that SOX2OT-7 is almost the most abundant form of SOX2OT transcript variants in the examined cancer cell lines particularly in NT2 teratocarcinoma cell line where its expression falls upon neuronal-like differentiation similar to SOX2 and OCT4A. We suggest that at least some of SOX2OT transcripts are significantly associated with cancer and stem cell related pathways. © 2015, Springer Science+Business Media Dordrecht

COVID-19: The Immune Responses and Clinical Therapy Candidates.

The pandemic of coronavirus disease 2019 (COVID-19), with rising numbers of patients worldwide, presents an urgent need for effective treatments. To date, there are no therapies or vaccines that are proven to be effective against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Several potential candidates or repurposed drugs are under investigation, including drugs that inhibit SARS-CoV-2 replication and block infection. The most promising therapy to date is remdesivir, which is US Food and Drug Administration (FDA) approved for emergency use in adults and children hospitalized with severe suspected or laboratory-confirmed COVID-19. Herein we summarize the general features of SARS-CoV-2's molecular and immune pathogenesis and discuss available pharmacological strategies, based on our present understanding of SARS-CoV and Middle East respiratory syndrome coronavirus (MERS-CoV) infections. Finally, we outline clinical trials currently in progress to investigate the efficacy of potential therapies for COVID-19

Amphotericin B potentiates the anticancer activity of doxorubicin on the MCF-7 breast cancer cells

Despite the improvements in cancer treatment, breast cancer still remains the second most common cause of death from cancer in women. Doxorubicin (DOXO) is widely used for cancer treatment. However, drug resistance limits the treatment outcome. Here, we investigated the toxicity of DOXO in combination with an antifungal agent amphotericin B (AmB) against the MCF-7 breast cancer cell line. The cell viability was measured using MTT assay. The apoptosis was studied by caspase-8 and caspase-9 activity measurements and DNA fragmentation was investigated by TUNEL assay. The combination of two drugs significantly increased the apoptotic index and the caspase-8 and caspase-9 activities in comparison to DOXO-treated cells. Our finding showed that pre-treatment of MCF-7 cells with AmB synergistically exerted the anticancer effect of DOXO through the caspase-dependent apoptosis manner. © 2017, Springer-Verlag Berlin Heidelberg

Sodium valproate modulates immune response by alternative activation of monocyte-derived macrophages in systemic lupus erythematosus

The anti-inflammatory role of macrophages in apoptotic cells (ACs) clearance is involved in Systemic Lupus Erythematosus (SLE) pathogenesis. The efferocytic capability of macrophages is altered by M1/M2 polarization. Histone deacetylase inhibitors (HDACi) are proposed to enhance the expansion of M2 macrophages. Sodium valproate (VPA) is an HDACi with different anti-inflammatory properties. Here, we aimed to investigate the effects of HDACi by VPA on the polarization of monocyte-derived macrophages (MDMs) and regulating the expression of anti-inflammatory cytokines in SLE. We studied the ex vivo alterations of MDMs among 15 newly diagnosed SLE patients and 10 normal subjects followed by ACs and VPA treatments. M1/M2 polarization was assessed by expression of CD86/CD163, IL1-β, IDO-1, and MRC-1 among treated and non-treated MDMs. We also evaluated the production of IL-10, IL-12, TGF-β1, and TNF-α cytokines in the cell culture supernatants. CD163 was overexpressed upon VPA treatment, while CD86 showed no significant change. IL1-β and IDO-1 genes were significantly downregulated, and the mRNA expression of MRC-1 was increased among VPA-treated MDMs of SLE patients. The anti-inflammatory cytokines (IL-10 and TGF-β1) were overproduced while TNF-α level was decreased in response to VPA. The population of classically activated macrophages was more prevalent among SLE patients and efferocytosis was defected. VPA could successfully enhance the anti-inflammatory immune response through alternative activation of MDMs in SLE patients. © 2017, International League of Associations for Rheumatology (ILAR)

Higher hepatic sterol-binding protein-2 gene expression and serum triglyceride level in methimazole induced subclinical hypothyroid animal model

Subclinical hypothyroidism (SCH) is a thyroid disorder characterized with elevated TSH level. Recent investigations indicate association of SCH with cardiovascular complications and dyslipidemia. Here, we aimed to evaluate the dyslipidemia in rat model of SCH induced with methimazole (MM) administration. For SCH induction, MM was administrated using gavage or ad libitum in drinking water to rats. Serum TSH and T4 levels were measured using ELISA method. The cholesterol and triglyceride were measured in serum samples using colorimetric assays. Lipid accumulation in the liver tissue was assessed histologically using oil red-o staining method. Gene expression alteration of the lipid hemostasis associated genes including acetyl CoA carboxylase (ACC), fatty acid synthase (FASN), and sterol-binding protein 2 (SREBP-2) were measured using qRT-PCR method in hepatocyte of treated animals. Our findings showed that MM can successfully induce SCH in rats by gavage; however, MM administration in drinking water was not efficient. The serum samples from SCH rat showed higher triglyceride (P value = 0.005) and total cholesterol (P value > 0.05) level. The accumulation of lipid droplets found in larger size (P value < 0.0001) in hepatocyte of the SCH rat comparing to the control. The gene expression of ACC, FASN, and SREBP-2 (~1.5 fold change) was upregulated in SCH rats; however, the changes of SREBP-2 expression were significant (P value = 0.04). It can be concluded that MM delivery through drinking water is not efficient enough for SCH modeling when comparing to gavage delivery. Altogether, we found MM-gavage-induced SCH rats have higher triglyceride level in circulation with increased lipid droplets size and elevated SREBP-2 transcription factor gene expression in hepatocytes. © 2021, The Author(s), under exclusive licence to Springer-Verlag London Ltd., part of Springer Nature

Immunomodulation in systemic lupus erythematosus: Induction of M2 population in monocyte-derived macrophages by pioglitazone

Macrophages have recently gained attention in systemic lupus erythematosus (SLE) pathogenesis for their role in the anti-inflammatory clearance of apoptotic cells. The M1/M2 polarization of macrophages improves efferocytic capability. Peroxisome proliferator-activated receptor Î3 is proposed to function in the expansion of the M2 subpopulation. Pioglitazone is a peroxisome proliferator-activated receptor Î3 agonist with a variety of anti-inflammatory effects. In this paper, we investigated the ex vivo alterations of monocyte-derived macrophages of 15 newly diagnosed SLE patients and 10 normal subjects triggered by apoptotic cells among SLE patients following pioglitazone treatment. The phagocytosis capacity of macrophages and M1/M2 polarization (CD86/CD163) was evaluated. The supernatants were also analyzed for the expression of interleukin (IL)-10, IL-12, transforming growth factor β1 and TNF-α. The mRNA expression of IL-1β and mannose receptor C-type 1 were also quantified among treated and non-treated monocyte-derived macrophages. We found that efferocytosis is defective among monocyte-derived macrophages of SLE patients and might be a major underlying mechanism involved in the sustained inflammation. Pioglitazone could enhance alternative activation of monocyte-derived macrophages and consequently immunomodulation in these patients. © 2017 Author(s)

The antifibrotic effects of naringin in a hypochlorous acid (HOCl)-induced mouse model of skin fibrosis

Objectives: Fibrosis is a chronic inflammation caused by the loss of innate compensational mechanisms. Naringin (NR) is a flavonoid with antineoplastic and anti-inflammatory effects. Here, we aimed to investigate the antifibrotic effects of NR and underlying mechanisms in a Hypochlorous acid (HOCl)-induced mouse model of skin fibrosis. Materials and methods: A total of 24 six-week-old female BALB/c mice were randomly allocated into five groups: HOCl, Sham, PBS, HOCl + NR and DMSO and selected skin regions were treated for 6 weeks, until sacrifice. The histopathologic and collagenesis of skin resections were analyzed using H&E and PR staining. The mRNA levels of COL1, COL3 and αSMA genes were quantified. Serum samples were also used to evaluate TGF-β levels and LDH activity. Results: HOCl could increase the relative collagen content, while NR administration on HOCl-treated biopsies decreased collagenesis. COL1, COL3 and αSMA mRNA levels were significantly increased among HOCl-treated skin samples, while NR treatment could decrease these mRNA levels of genes to the extent equal to the levels in the Sham group. Similarly, Naringin-treated samples could decrease TGF-β levels. Conclusions: We demonstrated that Naringin could exert protective effects against fibrotic complications of HOCL in skin tissue in vivo, by reducing the collagenesis and decreasing the levels of fibrosis-associated genes. © 2022 Informa UK Limited, trading as Taylor & Francis Group

SOX2OT knockdown derived changes in mitotic regulatory gene network of cancer cells

Background: SOX2 overlapping transcript (SOX2OT) is a long non-coding RNA, over-expressed in human tumor tissues and embryonic cells. Evidences support its function in the cell cycle; however there is no clear mechanism explaining its function in cell proliferation regulation. Here we investigated cancer cell response to SOX2OT knockdown by RNA sequencing. Methods: SOX2OT expression was inhibited by siRNA in two cancer cell lines (A549, U-87 MG), then the RNA of treated cells were used for the cDNA library synthesis and RNA sequencing. The differentially expressed genes were used for functional enrichment and the gene expression network was analyzed to find the most relevant biological process with SOX2OT function. Furthermore, the expression change of candidate genes was measured by qRT-PCR for more confirmation and the cell cycle was monitored by PI staining. Results: Our findings showed that SOX2OT knockdown affects the cellular gene expression generally with enriched cell proliferation and development biological process. Particularly, the cell cycle and mitotic regulatory genes expression including: CDK2, CDK2AP2, ACTR3, and chromosome structure associated genes like SMC4, INCENP and GNL3L are changed in treated cancer cells. Conclusion: Our results propound SOX2OT association with cell cycle and mitosis regulation in cancer cells. © 2018 The Author(s)

Effects of Raspberry Fruit (Rubus anatolicus (focke) foke ex hausskn) Hydroalcoholic Extract on Blood Glucose, Lipid Profile and Oxidative Stress Markers in Streptozotocin-diabetic Rats

Background and Objectives: Diabetes is one of the most common endocrine disorders worldwide. Rubus anatolicus is rich in polyphenolic compounds that can protect individual from various chronic diseases such as diabetes. This study was carried out to investigate effects of hydroalcoholic extract of raspberry fruit Rubus anatolicus (focke) on blood glucose levels, lipid profiles and oxidative stress markers in streptozotocin-induced diabetic rats. Materials & Methods: In this study, 32 male Wistar rats weighing 150�200 g were used. Diabetes was induced in rats using streptozotocin and diabetic animals were then treated with raspberry fruit hydroalcoholic extract for 14 days. Rats were anesthetized using xylazine and ketamine mixture and sacrificed. Then, whole blood samples were collected from the animal hearts to assess their serum glucose levels, lipid profiles and oxidative markers. The p-value < 0.05was considered as significant. Results: Results of the present study showed that raspberry fruit extracts significantly decreased serum glucose levels, triglycerides, cholesterol, low-density lipoproteins and malondialdehyde levels in diabetic rats, compared to the diabetic control rats (p < 0.05). Moreover, serum low-density lipoproteins level, total antioxidant capacity and superoxide dismutase activity significantly increased in diabetic rats, compared to diabetic control rats (p < 0.05). Conclusion: The present study showed that fruit extract raspberry included good effects on blood glucose levels, lipid profiles and oxidative stress conditions in diabetic rats. © 2022, National Nutrition and Food Technology Research Institute. All rights reserved