Pleckstrin Homology Domain Leucine-rich Repeat Protein Phosphatase Acts as a Tumor Suppressor in Oral Squamous Cell Carcinoma.

The pleckstrin homology domain leucine-rich repeat protein phosphatase (PHLPP) family has been found to have both tumor-suppressor and oncogenic properties across various types and locations of cancer. Given that PHLPP has not been previously studied in oral squamous cell carcinoma (SCC), we conducted an assessment of the expression of both its isoforms in oral SCC tissues and cell lines and compared these findings to their corresponding normal counterparts. In addition, we assessed the relationship between PHLPP and clinicopathological factors and patient survival. Quantitative real-time polymerase chain reaction was used to detect the mRNA levels of PHLPP1 and PHLPP2 in cancerous and normal cell lines in addition to 124 oral SCC and noncancerous adjacent epithelia (N = 62, each). Correlations between their expression rate and clinicopathological parameters were further evaluated in 57 patients. Data were statistically analyzed with t test and paired t test, analysis of variance, Mann-Whitney U , and Cox Regression tests ( P < 0.05). We found significantly lower levels of both PHLPP isoforms in oral SCC tissues compared with noncancerous epithelia ( P < 0.001, for both). However, in the cell lines, this difference was significant only for PHLPP1 ( P = 0.027). The correlation between the two isoforms was significant only in cancerous tissues ( P < 0.001). None of the clinicopathologic factors showed significant associations with either of the isoforms and there was no correlation with survival. We showed for the first time that PHLPP1 and PHLPP2 act as tumor suppressors in oral SCC at the mRNA level. The regulation of their mRNA appears to be different between normal and cancerous tissues.

Efficacy of synthesized cubic spirulina platensis photosensitizer in anticancer photodynamic therapy: An in vitro study.

Photodynamic therapy (PDT) is an option in select cancer management. Photosensitizers derived from natural sources can offer additional health benefits and play a crucial role in enhancing the efficacy of PDT in cancer treatment. We herein synthesized a cubic form of spirulina platensis (SP) and compared its anticancer-PDT efficacy with the naturally-occurring microhelical SP (MSP) and phycocyanin (Pc) against a tongue cancer cell-line and fibroblast cells. Cubic SP (CSP) was synthesized and characterized using standard analyses. CAL-27 and HGF cell-lines were incubated at different concentrations with each photosensitizer and were irradiated with 635 nm diode-laser. The viability, cellular-uptake, apoptosis and oxidative stress potential were quantitatively analyzed and statistically compared at P<0.05. Our results demonstrated that all three photosensitizers were non-toxic to normal cells before laser irradiation. In CAL-27, viability significantly decreased after PDT in all photosensitizer groups (P<0.05). Whereas, in HGF, Pc exhibited phototoxicity after laser irradiation (P=0.032). Cell-death was mainly apoptotic in Pc and CSP, but necrotic in MSP. Cellular-uptake was significantly higher in Pc, but was similar in MSP and CSP. Increase in reactive oxygen species was significantly higher in the Pc group compared to both SPs (P<0.05). We concluded that both SPs were safe and efficient photosensitizers for anticancer-PDT. CSP exhibited predominant and significant apoptotic death in CAL-27 and HGF cell-lines, while MSP mainly induced necrotic cell death. Despite the good photosensitizing performance of Pc, its use in higher concentrations should be considered with caution, due to the reduced viability that occurred following its use in PDT.

Diminution of STING Expression in the Blood of Patients with Severe or Critical COVID-19 Pneumonia.

INTRODUCTION: Cytokine storm and critical COVID-19 pneumonia are caused in at least 10% of patients by inborn errors of or auto-Abs to type I IFNs. The pathogenesis of life-threatening COVID-19 pneumonia in other patients remains unknown. METHODS: This study was conducted at Masih Daneshvari Hospital, Tehran, Iran. In the period of study, 75 confirmed cases of COVID-19 with presentations ranging from mild upper respiratory tract infection to lower respiratory tract infection, including moderate, severe, and critical disease, were recruited. Expression of STING mRNA was measured in peripheral blood mononuclear cells (PBMCs) and compared between patients with different severity and outcome. RESULTS: There was a significant negative correlation between age and STING expression level (p value = 0.010). Patients with "severe to critical" illness had a 20-fold lower STING expression level compared to the "mild to moderate" group (p value = 0.001). Also, the results showed lower expressions of STING in the patients admitted to the ICU (p value = 0.015). Patients who finally died had lower expression of STING at the time of sampling (p value = 0.041). CONCLUSION: STING mRNA expression in PBMCs was significantly lower in older COVID-19 cases, the patients with more severe illness, who needed intensive care, and who eventually died.

The role of angiotensin I converting enzyme insertion/deletion polymorphism in the severity and outcomes of COVID-19 patients.

The pandemic of coronavirus disease in 2019 has led to a global crisis. COVID-19 shows distinct clinical manifestations of the severity of symptoms. Numerous patients with no associated risk factors demonstrate acute respiratory distress syndrome (ARDS). The role of genetic factors in determining the severity and outcome of the disease remains unresolved. The purpose of this study was to see if a correlation exists between Angiotensin I Converting Enzyme (ACE) insertion/deletion (I/D) polymorphism and the severity of COVID-19 patients' symptoms. 120 COVID-19 patients admitted to Masih Daneshvari Hospital in Tehran with their consent to participate entered the study. Based on the World Health Organization classification, patients were divided into moderate and severe groups, which were primarily affected by O2 saturation levels. The effects of the patients' ACE insertion/deletion polymorphism, background disease, Angiotensin receptor blocker (ARB) drug consumption, and demographic parameters on the severity risk were calculated statistically. The ACE D allele was associated with an increased risk of disease severity (OR = 6.766, p = 0.012), but had no effect on mortality.

The potential key genes and pathways associated with Wilms tumor in quest of proper candidates for diagnostic and therapeutic purposes.

To designate the probable most important differentially expressed genes and genetic pathways in Wilms tumor and assess their expression and diagnostic potential by RT-PCR and statistical analysis. Systematic review of the literature and various bioinformatics analysis was carried out to gather and narrow down data. The expression of end-resulting genes was compared in Wilms tumor and normal tissue samples using RT-PCR. Statistical tests reported the diagnostic accuracy of genes and their correlation with clinicopathological features. Four genes including CDH1, NCAM1, EGF, and IGF2 were designated. The panel combining them has 100% sensitivity and specificity in differentiating tumors from normal tissue. Eight pathways, most involved in cell-cell and cell-basal matrix junction interactions, were found to be associated with disease pathogenesis. The suggested genes should undergo further evaluation to be validated as diagnostic biomarkers. Further research on the eight proposed pathways is recommended.

Distinctive expression of DNA replication factors in squamous cell carcinomas of the lip, face and oral cavity.

OBJECTIVE: Uncontrolled proliferation and aberrations in cell-cycle progression are fundamental issues in cancer. In this study we aimed to determine and compare deoxyribonucleic acid (DNA) replication licensing factors at the mRNA and protein levels among squamous cell carcinomas (SCCs) of the lip, facial-skin and oral cavity. MATERIALS AND METHODS: A total of 103 lip, oral and face SCCs were immunohistochemically stained with MCM2 (mini-chromosome maintenance 2), geminin, and ki67, and their labeling-indices were calculated. Also, 57 SCCs from the same regions along with their adjacent normal tissues underwent quantitative reverse transcription-polymerase chain reaction analysis. RESULTS: All three proteins were overexpressed in the studied SCCs, but only geminin (P = 0.004) showed significant difference among the three regions, with higher levels in oral SCCs compared to lip (P = 0.005) and skin (P = 0.024) tumors. Geminin expression did not differ between skin- and lip-SCCs (P = 0.822). MCM2/ki67 ratio was higher in oral- compared to skin-neoplasms (P = 0.039), but no difference was found in geminin/ki67 among the SCC-subsites. There were significant differences in MCM2 and geminin mRNA between carcinomatous- and normal-tissues in all tumors, but not among the three locations. CONCLUSION: MCM2 and geminin are involved in the tumorigenesis of lip, face and oral SCC at both mRNA- and protein-levels. Geminin may have a role in the site-specific biologic behavior of SCC. Skin SCCs had the highest proportion of licensed non-proliferating cells, while actively proliferating cells were more prominent in oral tumors. Regarding DNA replication, lip SCCs seem to be closer to skin tumors compared to their oral counterparts.

The effect of photodynamic therapy on head and neck squamous cell carcinoma cell lines using spirulina platensis with different laser energy densities.

BACKGROUND: Considering the anti-cancer properties of spirulina platensis (S. platensis), we aimed to investigate the effectiveness of this algae as a novel natural photosensitizer for photodynamic therapy (PDT) against oral and hypopharyngeal cancer cells. The appropriate laser energy density to apply during PDT was also determined. METHODS AND MATERIALS: CAL-27, FaDu and HGF cell lines were exposed to S. platensis with concentrations of 0.3 g/l and 0.6 g/l and were irradiated with 635 nm diode laser using 2, 4, 12, and 24 J/cm2 energy densities with constant power. MTT assay was performed to investigate cell viability and cytotoxicity after 24 h. The results were analyzed using two-way ANOVA and post hoc Tukey tests (P-value<0.05). RESULTS: survival rate in CAL-27 (P-Value<0.001) and FaDu (P-Value<0.001) cell lines were significantly different following irradiation with various laser energy densities. Different concentrations of S. platensis had no significant effect on the viability of CAL-27 cells (P-Value=0.158) and FaDu cells (P-Value=0.072) and showed no significant cytotoxicity against HGF cells, with or without laser. CONCLUSION: S. platensis could be considered as a novel safe and effective natural photosensitizer for cancer PDT with no cytotoxic effect on normal cells. When combined with laser using appropriate energy densities, it has the ability to induce death in oral and hypopharyngeal cancer cell lines.

Dual l-Carnosine/Aloe vera Nanophytosomes with Synergistically Enhanced Protective Effects against Methylglyoxal-Induced Angiogenesis Impairment.

Microvascular complications are among the major outcomes of patients with type II diabetes mellitus, which are the consequences of impaired physiological functioning of small blood vessels and angiogenic responses in these patients. Overproduction and accumulation of methylglyoxal (MGO), a highly reactive dicarbonyl byproduct of glycolysis pathway, has been acclaimed as the main inducer of impaired angiogenic responses and microvascular dysfunction in diabetic patients with uncontrolled hyperglycemia. Hence, an effective approach to overcome diabetes-associated microvascular complications is to neutralize the deleterious activity of enhanced the concentration of MGO in the body. Owing to the glycation inhibitory activity of Aloe vera whole extract, and capability of l-carnosine, an endogenous dipeptide, in attenuating MGO's destructive activity, we examined whether application of a combination of l-carnosine and A. vera could be an effective way of synergistically weakening this reactive dicarbonyl's impaired angiogenic effects. Additionally, overcoming the poor cellular uptake and internalization of l-carnosine and A. vera, a nanophytosomal formulation of the physical mixture of two compounds was also established. Although l-carnosine and A. vera at whole studied combination ratios could synergistically enhance viability of human umbilical vein endothelial cells (HUVECs) treated with MGO, the 25:1 w/w ratio was the most effective one among the others (27 ± 0.5% compared to 12 ± 0.3 to 18 ± 0.4%; F (4, 15) = 183.9, P < 0.0001). Developing dual nanophytosomes of l-carnosine/A. vera (25:1) combination ratio, we demonstrated superiority of the nanophytosomal formulation in protecting HUVECs against MGO-induced toxicity following a 24-72 h incubation period (17.3, 15.8, and 12.4% respectively). Moreover, 500 µg/mL concentration of dual l-carnosine/A. vera nanophytosomes exhibited a superior free radical scavenging potency (63 ± 4 RFU vs 83 ± 5 RFU; F (5, 12) = 54.81, P < 0.0001) and nitric oxide synthesizing capacity (26.11 ± 0.19 vs 5.1 ± 0.33; F (5, 12) = 2537, P < 0.0001) compared to their physical combination counterpart. Similarly, 500 µg/mL dual l-carnosine/A. vera nanophytosome-treated HUVECs demonstrated a superior tube formation capacity (15 ± 3 vs 2 ± 0.3; F (5, 12) = 30.87, P < 0.001), wound scratch healing capability (4.92 ± 0.3 vs 3.07 ± 0.3 mm/h; F (5, 12) = 39.21, P < 0.0001), and transwell migration (586 ± 32 vs 394 ± 18; F (5, 12) = 231.8, P < 0.001) and invasion (172 ± 9 vs 115 ± 5; F (5, 12) = 581.1, P < 0.0001) activities compared to the physical combination treated ones. Further confirming the proangiogenic activity of the dual l-carnosine/A. vera nanophytosomes, a significant shift toward expression of proangiogenic genes including HIF-1α, VEGFA, bFGF, KDR, and Ang II was reported in treated HUVECs. Overall, dual l-carnosine/A. vera nanophytosomes could be a potential candidate for attenuating type II DM-associated microvascular complications with an impaired angiogenesis background.

Evaluation of the Relationship Between Expression of Villin and Gelsolin Genes and Axillary Lymph Node Metastasis in Patients with Breast Cancer.

BACKGROUND & OBJECTIVE: Nowadays, actin-binding proteins such as Villin and Gelsolin have been considered to be associated with aggressive tumors. This study mainly aims to determine the relationship between Gelsolin and Villin genes expression and metastasis of axillary lymph nodes in patients with breast cancer. METHODS: The included population consisted of 40 confirmed cases of female breast cancer (including 20 patients with breast cancer along with axillary lymph node metastasis and 20 patients without axillary lymph node metastasis). Expression of Villin and Gelsolin genes was evaluated using Real-time PCR and pre-designed primers. RESULTS: The mean expression level of Villin in groups with and without axillary lymph node metastasis was 3.33±1.35 and 0.87±0.88, respectively (P<0.001). The mean Gelsolin expression levels in both groups (with and without axillary lymph node metastasis) were 4.13±2.40 and 1.00±0.35, respectively (P<0.001). The significant relationships were independent of individuals' age. CONCLUSION: Patients with axillary lymph node metastasis may express significant higher level of Villin and Gelsolin genes.

A Novel Coronavirus Disease (COVID-19): a Review of Host Cell Signaling Pathways.

Coronaviruses (CoVs) are the largest group of positive-sense RNA viruses. By increasing our understanding of the interactions between CoVs and the host innate immune system, we can evaluate the development and persistence of inflammation in the lungs and reduce the risk of CoV-induced lung inflammation with a new group of genetic variants. Here, we aim to discuss some recent changes in host cell factors that may be used by CoV to promote the proliferation cycle. We also discuss different host cell signaling pathways that can be considered in the host-pathogen interactions at the molecular level. The pandemic of coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has created new challenges for the cultural, economic, and health infrastructures. Therefore, it is important that healthcare systems and physicians recognize a global integrated framework for monitoring the progression of COVID-19 to develop targeted therapies that can potentially save human lives.

Association of High Expression Levels of SOX2, NANOG, and OCT4 in Gastric Cancer Tumor Tissues with Progression and Poor Prognosis.

BACKGROUND: Expression of the essential regulator genes, SOX2, NANOG, and OCT4, so-called as stemness factors, is prerequisite for the tumorigenic capability of cancer stem cells (CSCs) and their potential role in the formation and progression of various human cancers. METHODS: In this study, the expression levels of SOX2, NANOG, and OCT4 were quantified by a qRT-PCR method in 100 gastric cancer tumor tissues vs the paired adjacent normal tissues. Then, the relationship between the expression of the three genes in gastric cancer tumor tissues and the clinicopathological characteristics and overall survival of patients was investigated. RESULTS: Higher expression levels of SOX2, NANOG, and OCT4 were found in gastric cancer tumor tissues compared with those in paired adjacent normal tissues (P = 0.0001). Overexpression of the mentioned genes in gastric cancer tumor tissues was resolved to be significantly associated with tumor size (P < 0.05), TNM stage (P = 0.001), tumor grade (P < 0.01), and shortened overall survival time (P = 0.0001). CONCLUSIONS: These findings indicted that the stemness factors SOX2, NANOG, and OCT4 are significantly overexpressed in gastric cancer and may serve as potential biomarkers of gastric cancer progression and prognosis.

Association of MAPK and its regulatory miRNAs (603, 4301, 8485, and 4731) with the malignant transformation of oral lichen planus.

Oral lichen planus (OLP) is a potentially malignant oral lesion that may transform into oral squamous cell carcinoma (OSCC). The purpose of this study was to assess the level of expression of MAPK/ERK1/2 gene, and microRNA (miR)-603, 4301, 8485, and 4731 in the MAPK signaling pathway in OLP and OSCC lesions. This case-control study evaluated 26 OSCC, 20 OLP and 20 healthy control tissue specimens. After RNA extraction, the respective miRNA and MAPK/ERK1/2 mRNA levels were assessed by quantitative reverse transcription polymerase chain reaction (RT-PCR). Significant upregulation of MAPK/ERK1/2 gene was noted in the OLP and OSCC specimens compared with healthy controls (p < 0.001). The expression level of miR-4731 was significantly lower in the OLP and OSCC specimens than in the healthy specimens (p < 0.001). The expression of MiR-603 was the lowest in OLP, followed by OSCC and then the control group (p < 0.001). No significant difference was found in miR-4801 levels between OSCC and OLP specimens compared with healthy controls (p = 0.43 and p = 0.86, respectively). In addition, a non-significant decrease in miR-8485 levels was noted in the OSCC and OLP specimens compared with healthy controls (p = 0.98 and p = 0.61, respectively). A significant decrease in level of miR-603 was noted in OLP compared with OSCC group (p < 0.001). The miR-4801 and miR-8485 expression levels were directly correlated with MAPK/ERK1/2 mRNA expression (p = 0.01). Higher expression level of MAPK/ERK1/2, miR-603, miR-4801, and miR-4731, and lower expression level of miR-8485 were correlated with significantly lower overall survival rate in OSCC patients. The increased expression of MAPK/ERK1/2 and decreased expression of miR-603 and miR-4731 are associated with greater risk of OLP malignant transformation and poor histopathological characteristics of OSCC.

Relative Expression of OCT4, SOX2 and NANOG in Oral Squamous Cell Carcinoma Versus Adjacent Non- Tumor Tissue.

Objective: Oral squamous cell carcinoma (OSCC) accounts for over 90% of oral neoplasms. Finding molecular markers for predicting prognosis is a high priority. The core transcription factors, OCT4, SOX2, and NANOG that regulate embryonic stem cell pluripotency have been implicated in progression of various malignancies. The predictive value of these markers and their role in the development of OSCC is still controversial. In this study, we therefore evaluated their expression in OSCCs and adjacent non-tumor tissue. Methods: A total of 60 frozen tumor and adjacent non-tumor tissue samples from 30 patients with OSCC were examined using quantitative reverse transcription polymerase chain reaction (qRT-PCR). Clinical and pathological data of patients including tumor stage, lymph node metastasis and tumor grade were also recorded. Results: Expression of SOX2 was significantly higher in adjacent non-tumor as compared to tumor tissue (P=0.04). No statistically significant differences were found for expression of OCT4 (P=0.50) and NANOG (P=0.68). Also, there was no significant association between expression of OCT4, SOX2, and NANOG and clinical or pathological data (P>0.05), although slightly higher values were noted in patients without lymph node metastasis. Conclusion: Based on the present data, decreased expression of SOX2 is correlated with carcinogenesis in the oral cavity and development of OSCC.

PTEN and miR-26b: Promising prognostic biomarkers in initiation and progression of Oral Squamous Cell Carcinoma.

BACKGROUNDS: Oral Squamous Cell Carcinoma (OSCC) is the most common malignancy of the oral cavity. Phosphatase and TENsin homolog (PTEN) is a well-known tumor suppressive gene regulated by several biomarkers including a small single-stranded molecule, microRNA26b (miR-26b). Here, we studied the expression of PTEN and miR-26b in OSCC specimens in comparison with adjacent normal mucosa. METHODS: The expressions of PTEN and miR-26b genes were evaluated at mRNA level in OSCC and adjacent normal fresh frozen tissues in 49 patients using Quantitative Real-Time PCR and analyzed their associations with clinicopathological factors. RESULTS: The expression level of PTEN was significantly lower in OSCC specimens comparing with adjacent normal tissues (P-value = 0.000). The expression of PTEN was associated with T stage (P-value = 0.006) and N stage (P-value = 0.043). A nonsignificant decrease in miR-26b expression level was also observed in OSCC tissues. Additionally, in patients with more aggressive tumoral behavior, including vascular invasion (P-value = 0.012) and positive N stage (P-value = 0.02), significant decreases were found. CONCLUSIONS: These findings suggest that inactivation of PTEN may have an impact on initiation and progression of OSCC. Additionally, miR-26b might have a tumor suppressive role in OSCC.