Objectives:Oxidative stress promotes the initiation and progression of diabetic complications. Fibrinogen is a thrombotic factor proposed as a biomarker for prognostic assessment, as a marker for tracking vascular complications, and as a therapeutic target in diabetes. Aminoguanidine, an inhibitor of advanced glycation end products and inducible nitric oxide synthase, has been shown to reduce oxidative stress and related vascular injury. However, the underlying processes still need to be entirely elucidated. This study investigated the effects of aminoguanidine on oxidative stress biomarkers and fibrinogen gene expression in diabetic rats. Materials and Methods:Twenty-four male rats, matched for body weight and age, were randomly assigned to five groups: (intended n = 6 per group). Due to mortality during the study, the final group sizes were: healthy controls (n = 5), untreated diabetic controls (n = 4), and diabetic rats treated with aminoguanidine at 50, 100, or 200 mg/kg (n = 6, 4, and 5, respectively). Diabetes mellitus was induced by intraperitoneal injection of streptozotocin (50 mg/kg) in rats fasted for 12 h. One week later, diabetic rats received aminoguanidine for 28 days. The liver was selected for analysis due to its key role in oxidative stress and metabolism. Malondialdehyde (MDA) and ferric-reducing ability of plasma (FRAP) were measured in liver tissue as markers of oxidative stress, and fibrinogen gene expression was assessed by real-time polymerase chain reaction. Results:Aminoguanidine at 50 mg/kg significantly reduced fibrinogen gene expression (p = 0.047). Hepatic MDA levels decreased at all doses (50, 100, and 200 mg/kg, p < 0.05), while FRAP levels increased significantly at 50 and 100 mg/kg (p < 0.05). Conclusion:Aminoguanidine may attenuate oxidative stress and fibrinogen gene expression in the livers of diabetic rats.

Oral squamous cell carcinoma (OSCC) is an aggressive epithelial malignancy with limited therapeutic options and high recurrence rates. C-Phycocyanin, a phycobiliprotein derived from Spirulina platensis, exhibits promising anticancer properties by modulating various molecular pathways. This study aimed to investigate the cytotoxic and pro-apoptotic effects of C-Phycocyanin on human oral cancer cells (KB) and compare its safety profile on normal endothelial cells (HUVECs). Cell viability was assessed using the MTT assay in both KB and HUVEC cell lines following treatment with different concentrations of C-Phycocyanin. Apoptotic features were evaluated by DAPI nuclear staining and quantified using Annexin V/PI-based flow cytometry. The expression of apoptosis-related genes (P53, Bax, Bcl-2) and signaling molecules involved in the Akt/PTEN and MAPK (ERK2) pathways was analyzed by semi-quantitative RT-PCR. C-Phycocyanin significantly inhibited the viability of KB cells in a dose-dependent manner while exerting minimal cytotoxic effects on HUVEC cells. DAPI staining revealed nuclear fragmentation and chromatin condensation in KB cells. Flow cytometry confirmed apoptosis induction. RT-PCR analysis showed upregulation of P53 and Bax and downregulation of Bcl-2, suggesting activation of the intrinsic apoptotic pathway. In addition, C-Phycocyanin inhibited Akt and upregulated PTEN and MAPK (ERK2) pathway components. C-Phycocyanin effectively induces apoptosis in oral cancer cells by modulating key molecular pathways, with minimal toxicity on normal cells. These results support its potential as a natural and selective therapeutic agent for OSCC, meriting further in vivo validation.

Breast cancer (BC) is the second cancer-dyeing reason among women worldwide. Surgical, radiation, and medicine-based approaches are the current methods for BC treatment; however, they did not overcome metastasis BC robustly. Finding treatments to improve the survival rate appears to be an essential attitude. Telomer is a fundamental part of the linear eukaryote chromosome. Despite the low-telomerase activity in somatic cells, it has been reported that more than 90

Introduction. Diabetic nephropathy is one of the most common severe symptoms of diabetes mellitus. Hyperglycemia can lead to tissue damage and inflammation due to mediators such as receptor for advanced glycation end-products (RAGE). Therefore, in this study, we aimed to investigate the association between the G82S polymorphism of the RAGE gene and diabetic nephropathy in diabetic patients . Methods. In this case-control study, 356 participants (158 men and 198 women) of Asian race, aged 45 to 65 years, who were diagnosed with type 2 diabetes mellitus based on their fasting plasma glucose levels were enrolled. DNA was isolated from the participants' blood samples and genotyped using TETRA-Primer ARMS-PCR. Serum protein concentration of soluble RAGE (sRAGE) was also determined by enzyme-linked immunosorbent assay (ELISA). Results. Although we found differences in genotyping of participants between homozygous AA and GG and heterozygous GA in the studied groups, the differences were not significant (P = .568). In addition, we found no significant correlation between the G82S polymorphism of RAGE and the development of diabetic nephropathy. Serum levels of sRAGE were only slightly decreased in patients with diabetic nephropathy compared with diabetic patients (P > .05). Conclusion. The results of this study indicate no significant association between the G82S polymorphism in the gene RAGE and the development of diabetic nephropathy. Serum levels of sRAGE were only slightly decreased in patients with diabetic nephropathy compared to diabetic patients without nephropathy. Therefore, the study suggests that there is probably no association between the G82S polymorphism in the gene RAGE and the development of diabetic nephropathy.

Integrin αvβ3, a primary cell-adhesion receptor, plays a crucial role in various biological processes, including angiogenesis, pathological neovascularization, and tumor metastasis. Its expression increases during tumor angiogenesis. The insulin-like growth factor 1 receptor (IGF1R) is a transmembrane protein that stimulates vital signaling pathways, promoting cancer cell growth, survival, and metabolism. Notably, a crucial step in insulin-like growth factor (IGF) signaling in non-hematologic cancers is IGF1 binding to integrins αvβ3, followed by the formation of the integrin-IGF1-IGF1R complex that dual silencing of these genes could enhance therapeutic specificity on MCF-7 breast cancer cells. In the current study, mPEG-PCL-DDAB nanoparticles (NPs) were synthesized by hybridizing methoxy polyethylene glycol-poly caprolactone (mPEG-PCL) with the cationic lipid dimethyldioctadecylammonium bromide (DDAB). These nanoparticles were utilized as a carrier of siRNAs for the dual silencing of integrin αvβ3 receptor and IGF1R. The results from the in vitro study indicate that this nanoparticle with high encapsulation efficiency of siRNAs, dramatically induces the process of apoptosis, gene silencing, and cell cycle arrest in MCF-7 tumor cell lines. In general, the article represents a significant advancement in MCF-7 breast cancer cell research by developing a novel nanoformulation as a therapeutic approach involving integrin αvβ3 receptor and IGF1R.

OBJECTIVE:The adipokine Chemerin and the retinoic acid receptor responder 2 (RARRES2) gene have been associated with an increased incidence of obesity, insulin resistance, endothelial dysfunction, type 2 diabetes mellitus, and coronary artery disease. The impact of RARRES2 rs17173608 gene polymorphism on acute myocardial infarction and Chemerin levels has not yet been entirely elucidated. This study aimed to assess the association of RARRES2 rs17173608 gene polymorphism and serum Chemerin with acute myocardial infarction (AMI) and its risk factors in an Iranian population.METHODS:In this case-control study, 134 AMI patients and 100 healthy controls were recruited from tertiary referral hospitals in Zanjan, Iran. Whole blood samples were collected for DNA extraction and Chemerin level determination. An enzyme-linked immunosorbent assay was used to quantify plasma levels of Chemerin. Tetra-primer amplification refractory mutation system-polymerase chain reaction and agarose gel electrophoresis techniques were used to detect gene polymorphism of Chemerin rs17173608 from DNA samples.RESULTS:Serum Chemerin levels were higher in the control group than in the AMI group. However, after adjusting for age, sex, and other risk factors, there was no significant association between serum Chemerin level and AMI occurrence. In the genotype analysis, 21.6% of patients had the TT genotype, and 78.4% had the TG genotype. The GG genotype was not detected in any patient. The genotype distribution of the healthy control group was 19.0% TT genotype, 80.0% TG genotype, and 1.0% GG genotype. Serum Chemerin levels in participants with TG genotype were statistically different between case and control groups.CONCLUSION:Serum Chemerin levels and RARRES2 rs17173608 gene polymorphism were not correlated with AMI occurrence after adjusting for AMI risk factors in Iranian patients. More research with a larger sample size and diverse ethnicities is needed to corroborate our findings.

Objectives: Advanced glycation end products (AGEs) play an important role in the development and progression of diabetic complications. The receptor for AGE (RAGE) is the ligand-binding site of AGE that initiates and accelerates the atherosclerotic process. Plasminogen activator inhibitor-1 (PAI-1) is a prothrombotic factor that has been proposed as a biological marker for prognostic assessment, monitoring of microvascular and macrovascular complications in diabetes. The purpose of this study is to investigate the effects of aminoguanidine on RAGE and PAI-1 expression levels in the liver of streptozotocin-induced diabetic rats. Methods: Diabetes was induced in rats by intraperitoneal injection of streptozocin (STZ, 50 mg/kg). On day 3, diabetic rats were administered 50, 100, and 200 mg/kg/day of aminoguanidine. The expression of PAI-1 and RAGE in the liver tissue was evaluated using real-time PCR. Results: PAI-1 and RAGE gene expression levels were higher in the liver of the diabetic rats compared to the control group. Aminoguanidine at 50, 100, and 200 mg/kg decreased PAI-1 and RAGE gene expression in the liver (p<0.001 at all doses). However, these genes were downregulated only at a dose of 200 mg/kg in healthy rats (p<0.0001). In addition, hepatic AGE protein levels were significantly decreased following treatment of the diabetic rats with aminoguanidine (p<0.001). There was also a significant correlation between AGE protein concentration and the expression of PAI-1 and RAGE. Conclusion: In summary, the data of the present study suggest that aminoguanidine reduced the expression of PAI-1 and RAGE in the liver of the diabetic rats.

Evaluation of the relationship between ABCB11 gene polymorphism and the incidence of gallstones and related SGOT and SGPT biochemical factors in the Iranian population

Purpose This study aims to evaluate the effects of betanin on AMP-activated protein kinase (AMPK), Sirtuin1 (SIRT1) and Sirtuin6 (SIRT6) gene expression as well as the tumour necrosis factor alpha (TNF-alpha) and interleukin 6 (IL-6) cytokine release in peripheral blood mononuclear cells (PBMCs) of patients with coronary artery disease (CAD) and healthy controls. Design/methodology/approach PBMCs isolated from whole blood of 50 patients with CAD and 48 healthy subjects aged 45 to 60 years were treated with 10 and 20 mu M of betanin for 24 h. Real-time polymerase chain reaction was performed to assess gene expression levels of AMPK, SIRT1 and SIRT6. The supernatants of the cultured cells were used to assess the IL-6 and TNF-alpha protein levels by ELISA. Findings Treatment with both doses of betanin significantly increased AMPK, SIRT1 and SIRT6 expression in PBMCs of CAD patients compared to control non-treated cells (p < 0.05). In PBMCs of healthy subjects, only treatment with high dose of betanin showed significant increase in AMPK (p = 0.007), SIRT1 (p = 0.013) and SIRT6 (p = 0.024) expression compared to control non-treated cells. Betanin (20 mu M) also significantly decreased TNF-alpha and IL-6 concentrations in the culture supernatants of the CAD patients compared to control non-treated cells (p < 0.001). Originality/value Betanin could enhance AMPK, SIRT1 and SIRT6 gene expressions in PBMCs and represent a useful complementary treatment to reduce the proinflammatory status accompanied with CAD.

Introduction:Sphingosine 1 phosphate (S1P) is a product of the sphingosine kinase 1 (SphK1) enzyme. Increased S1P can lead to tissue fibrosis that is also one of the pathways for developing diabetic cardiomyopathy. Advanced glycation end products (AGEs) increase S1P in cells. The study is aimed at using aminoguanidine (AG) as an AGEs blocker drug to prevent diabetic cardiomyopathy. Materials and methods. 210 rats were enrolled in the study. Diabetes mellitus type-2 was induced, and rats were divided into AG treated diabetic and nondiabetic groups. The heart histology was assessed with Masson's trichrome and hematoxylin-eosin staining. Cardiac function was measured with transthoracic echocardiography. S1P level and SphK1 gene expression were measured by western-blot and RT-qPCR, respectively.Results:Results showed that S1P level increases in diabetes, and its augmentation in cardiac tissue with K6PC-5 leads to cardiac fibrosis. 50 and 200 mg/kg of AG prevented cardiac fibrosis, but 100 mg/kg had no significant preventive effect. AG suppressed the SphK1 gene expression and reduced the fibrotic effect of S1P. AG preserved cardiac function by keeping ejection fraction and fractional shortening within the normal range in diabetic rats.Conclusion:AG has a suppressor effect on SphK1 gene expression besides its AGEs blocker role. AG is a potential drug to use in diabetic patients for preventing the development of diabetic cardiomyopathy. Other drugs that have AGEs or S1P blocker effects are a good choice for diabetic cardiomyopathy prevention.

Insulin-like growth factor-1 receptor (IGF-1R) is expressed in malignant and normal breast tissue, and its intermittent activation by multiple IGF-1 signaling pathways leads to neoplasm cell proliferation, impaired apoptosis, increased survival, and resistance to cytotoxic therapeutic agents. Therefore, simultaneous suppression of the receptor and its cognate ligand would be a powerful promising strategy inhibiting malignant phenotypes of breast cancer cells. In the present study, Methoxypoly(ethylene glycol) - Poly(caprolactone) was hybridized with Dimethyldioctadecylammonium bromide (DDAB) cationic lipid (mPEG-PCL-DDAB) nanoparticles (NPs) and used as a carrier for simultaneous delivery of lycopene and insulin-like growth factor 1 receptor-specific lycopene encapsulated-mPEG-PCL-DDAB nanoparticle/siRNA to MCF-7 breast cancer cells. Then, the antitumor effects of this construct were evaluated in vitro. The results demonstrated that the synthesized mPEG-PCL-DDAB nanoparticle had suitable physicochemical properties. The use of mPEG-PCL-DDAB nanoparticle-loaded anti-insulin-like growth factor 1 receptor-siRNA and lycopene dramatically induced the process of apoptosis and arrested cell cycle in the MCF-7 tumor cell lines. In general, the findings of this study demonstrated the potency of mPEG-PCL-DDAB nanoparticles for dual delivery of siRNA, and lycopene in breast cancer cell lines followed the induction of apoptosis.

Abstract Systemic inflammation and oxidative burden in patients with type 2 diabetes mellitus (T2DM) causes deleterious cardiovascular outcomes. We sought to investigate the clinical antioxidative and anti-inflammatory effects of empagliflozin. Platelet function, oxidant and antioxidant biomarkers and pro-inflammatory agents at baseline and at 26 weeks were measured. A total of 95 patients (41.05% male, mean age 62.85 ± 7.91 years, mean HbA1c 7.89 ± 0.96%) with concomitant T2DM and coronary artery disease (CAD) were randomized (1:1) to receive empagliflozin (10 mg/daily) or placebo. Patients treated with empagliflozin had lower levels of interleukin 6 (IL-6) (adjusted difference (adiff): − 1.06 pg/mL, 95% CI − 1.80; − 0.32, P = 0.006), interleukin 1β (IL-1β) and high-sensitive C-reactive protein (Hs-CRP) (adiff: − 4.58 pg/mL and − 2.86 mg/L; P = 0.32 and 0.003, respectively) compared to placebo. There were elevations in super oxidase dismutase (SOD) activity, glutathione (GSHr), and total antioxidant capacity (TAC) with empagliflozin (adiff: 3.7 U/mL, 0.57 muM, and 124.08 mmol/L, 95% CI 1.36; 6.05, 0.19; 0.95, and 47.98; 200.18, P = 0.002, 0.004, and 0.002, respectively). While reactive oxygen species (ROS) improved significantly (adiff: − 342.51, 95% CI − 474.23; − 210.79, P < 0.001), the changes in catalase activity (CAT), malondialdehyde (MDA), or protein carbonyl groups (PCG) were not significant. Moreover, the P-selectin antigen expression on platelet surface was significantly reduced (adiff: − 8.81, 95% CI − 14.87; − 2.75, P = 0.005). Markers of glycemic status (fasting blood glucose, HbA1c, and HOMA-IR (homeostatic model assessment for insulin resistance) significantly improved (P < 0.001). Among patients with T2DM and CAD, 6-month treatment with empagliflozin can mitigate inflammation, platelet activity and oxidative stress and is associated with clinical cardiovascular benefits. Trial Registration Iranian Registry of Clinical Trials. www.IRCT.ir , Identifier: IRCT20190412043247N2. Registration Date: 6/13/2020. Registration timing: prospective

Background: Metabolic disturbances, including hyperlipidaemia, are risk factors for brain ischemia. Despite all research, no definitive treatment has yet been found for brain ischemia. Thus, this study investigated the effect of cinnamon on ischemic tolerance and the expression of matrix metalloproteinase (MMP) 2 and 9 genes, as well as levels of interleukin 6 (IL-6) and tumor necrosis factor α (TNF-α) proteins in rats receiving a high-fat diet (HFD). Methods: Rats were divided into control, sham, model, lovastatin, vehicle, and cinnamon 130 mg (Cin130) and 260 mg (Cin260). All groups, except for the control group, received a high-fat diet for 6 weeks. Then, the last four groups received the corresponding treatment for 6 weeks. The stroke was induced by middle cerebral artery occlusion (MCAO). Twelve hours later, the animals were evaluated for the extent of neurological defects, lipid profiles, tissue damage, and gene expression of MMP-2,9, as well as IL-6 and TNF-α protein level. Results: The results showed that Cin130 has been highly successful in improving neurological defects. Cin130 was effective in reducing serum cholesterol and triglyceride. Further, Cin260 effectively reduced serum LDL. Moreover, Cin130 and Cin260 could reduce tissue damage volume, blood-brain barrier (BBB) integrity, and down-regulation of IL-6, TNF-α proteins and MMP-2,9 levels. Conclusion: Overall, cinnamon, in addition to its lipid-lowering effect, may also have a neuroprotective effect that may be related to the down-regulation of MMPs and cytokines. The two may be independent events or related to a third event; in any case, appropriate experiments should be performed in future studies to determine whether altering the MMPs reduces the effectiveness of cinnamon.

Objective Pre-eclampsia (PE) is a serious pregnancy status characterized by high blood pressure. Although visfatin is usually associated with PE. Observational studies evaluating the relationship between circulating visfatin and pre-eclampsia have reported inconsistent results. We conducted this systematic review and meta-analysis to summarize published data on the association between visfatin and pre-eclampsia Methods Electronic databases PubMed, ISI web of science, EMBASE, Scopus and the Cochrane library were comprehensively searched for selection of eligible studies until January 5, 2020. A random-effects model and the generic inverse variance method were used for quantitative data synthesis. The assessment of study quality was performed using the e Newcastle-Ottawa scale and the Agency for Healthcare Research and Quality. Sensitivity analyses and prespecified subgroup were conducted to evaluate potential heterogeneity. Random-effects meta-regression was conducted to assess the impact of potential confounders on the estimated effect sizes. The protocol for this study was registered in PROSPERO (No. CRD42018105861) in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA). Results Thirteen studies comprising a total of 536 subjects were included in this meta-analysis. We observed that the pre-eclampsia risk is associated with a statistically significant elevation of visfatin level [SMD (1.33 mu g/l) (95% CI 0.37, 2.2)p = .007]. No significant publication bias was observed in the meta-analysis. Subgroup and sensitivity analyses indicated that the pooled effects size were affected by systolic blood pressure [SMD (1.82 mu g/l) 95% CI (0.94, 2.7),p < .05], gestational age [SMD (2.01 mu g/l) 95% CI (0.57, 3.4),p = .006], body mass index [SMD (1.6 mu g/l) 95% CI (0.37, 3),p < .05] and pregnancy trimesters[SMD (2.3 mu g/l) 95% CI (0.95, 3.7),p = .001]. Random-effects meta-regression showed a significant association of visfatin level with potential confounders including systolic blood pressure, gestational age and birth weight at delivery of pre-eclampsia patients. Conclusions Collectively, our data revealed that the increase of visfatin level can be associated with the risk of pre-eclampsia. However, further studies on pre-eclampsia populations are warranted for corroboration of our findings.

Stroke is the second leading cause of death and disability worldwide. The present study was designed to investigate the neuroprotective effects of polymeric micelles loaded with Edaravone on ischemia-induced in human neuroblastoma cell line (SH-SY5Y). Polymethoxy ethylene glycol-polylactic-co-glycolic acid (mPEG-bPLGA) was first synthesized, and its physicochemical properties were determined. Micelles were prepared by the nanoprecipitation method. The size of micelles was 155 +/- 2.5 nm with a surface charge of -15 +/- 0.94 and polydispersity index (PDI) of 0.11. The examination of the morphology of micelles performed by transmission electron microscopy (TEM) and atomic force microscopy (AFM) showed that the size of particles was less than 155 +/- 2.5 nm, and the morphology was spherical. The drug loading (DL) was determined about 13 +/- 1.4% and encapsulation efficiency was 65 +/- 0.79%. Drug delivery was analyzed using UV-VIS spectroscopy. The impact of different concentrations of free and micelle-containing Edaravone was performed on SH-SY5Y cells under normal and oxygen and glucose deprivation (OGD) conditions. Lactate dehydrogenase (LDH), nitric oxide (NO) activity, Intracellular reactive oxygen species (ROS) and apoptosis rate were significantly reduced in cells treated with edaravone loaded-micelle in addition to a significant reduction in the expression of pro-apoptotic gene (Bax) and increase in the expression of anti-apoptotic genes (HSP70 and Bcl-2). Consequently, according to the result of the study Edaravone-loaded micelles had a significant rate of neuroprotective effects than free Edaravone against ischemia-induced oxidative damage.

Background Continuing hyperglycemia causes and exacerbate oxidative stress. Betanin as the principal pigment of red beet root has antioxidant, anti-inflammatory, and anti-diabetic properties. The purpose of this study was to investigate the potency of betanin on antioxidant defense in STZ-induced diabetic rats’ livers. Methods STZ at a single dose of 60 mg/kg body weight was intraperitoneally injected and betanin (10, 20, and 40 mg/kg/day) was administered orally for 28 days. Malondialdehyde (MDA), total antioxidant capacity (TAC), protein carbonyl (PC) levels, and the enzyme activity of superoxide dismutase (SOD), catalases and glutathione peroxidases (GPx) were evaluated in the liver. Furthermore, gene expression of Nrf2 and mentioned antioxidant enzymes were measured by Real-time PCR. Results Betanin (10 and 20 mg/kg) significantly reduced PC levels and increased antioxidant enzyme activity in diabetic rats compared to the control diabetic group (P < 0.01). In comparison to the diabetic control group, all studied genes expression in diabetic rats were increased significantly with betanin at doses of 10 and 20 mg/kg (P < 0.02). The increase in gene expression at 20 mg/kg of betanin was significantly stronger than others (P < 0.015) except for the catalase (P = 0.201), that was almost the same. Moreover, treatment of diabetic rats with 20 mg/kg of betanin could significantly increase TAC levels (P < 0.05) and decrease MDA levels (P < 0.001) compared to diabetic control group. Conclusions Betanin could increase the antioxidant capacity of liver tissue associated with the Nrf2-mediated pathway in a dose-dependent manner.

Colorectal cancer (CRC) is the second leading cause of cancer death. Although Regorafenib showed survival benefits in patients with CRC, reports imply the recurrence of malignant phenotype resulting from chemotherapy. Evidence demonstrated that a5β1 integrin plays an important role in the Regorafenib treatment, which, may be led to resistance. In this study, the effects of /siRNA or/ and Quercetin loaded DDAB-mPEG-PCLnanoparticles could reverse this resistance phenotype in colon cancer cells in vitro.Regorafenib-resistant Ls-180 colon cancer cell line was developed by long-term exposure to Regorafenib. Quercetin and Regorafenib were separately encapsulated into mPEG-PCL micelles through the nano-precipitation method and characterized by DLS. Optimized doses of Quercetin and Regorafenib were used for combination therapy of resistant cells followed cytotoxicity study using MTT. Gene expression levels of the β1 subunit of integrin were determined by the real-time method of RT-PCR.Developed Regorafenib resistant LS-180 showed to have Regorafenib IC50 of 38.96 ± 1.72 µM whereas IC50 in non-resistant cells were 8.51 ± 0.29 µM, which meaningful was lower statistically compared to that of a resistant one. The β1 mRNA level of whole α5β1 integrin was significantly higher in the resistant cells compared to those of non-resistant ones. Gene expression levels in each siRNA-loaded nanoparticle and Quercetin-loaded one were lower than that in mock experiments. Finally, when these two types of nanoparticles were used to treat resistant cells, gene expression decrease of integrin indicated a greater effect that could be capable of reverse resistancy.Results of this study demonstrated another confirmation of involving integrins in cancer resistance following chemotherapy using Regorafenib. Also, it indicated how using siRNA targeting integrin could enhance the plant derivatives like Quercetin effects to reverse resistance in vitro.

BACKGROUND:The DNA mismatch repair (MMR) system is one of the molecular pathways involved in colorectal cancer (CRC) carcinogenesis that consists of several genes, including MLH1 (MutL homolog 1), MSH6 (MutS homolog 6), MSH2 (MutS homolog 2), and MSH3 (MutS homolog 3). The protein encoded by PMS2 (post-meiotic segregation 2) is also essential for MMR. Here, we address the correlation between immunohistochemical and transcriptional expression of PMS2 with the tumor grade and clinical stage of non-hereditary/sporadic CRC disease.METHODS:This study retrospectively analyzed 67 colorectal resections performed for 38 male and 29 female patients. Random biopsies were taken by a gastroenterologist from patients referring to three hospitals in the cities of Zanjan, Urmia and Qazvin (Iran) during 2017-2019. All specimens were examined and classified for localization of tumor, pathological stage and grade. The PMS2 protein expression was studied immunohistochemically and analysis of mRNA expression was performed in the same tissue sections.RESULTS:Immunohistochemistry and quantitative real-time polymerase chain reaction (PCR) analysis showed a decrease in PMS2 expression compared with paracancerous tissue (P<0.001), which correlated with tumor stage. In addition, reduced PMS2 expression was correlated with the tumor differentiation grade, underlining a connection between downregulation of PMS2 and progression of CRC. Comparing the PMS2 mRNA levels in different groups showed the following results: 0.92 ± 0.18 in patients with Stage I CRC tumor, 0.86 ± 0.38 in Stage Ⅱ, 0.50 ± 0.29 in Stage Ⅲ, and 0.47 ± 0.23 in Stage Ⅳ.CONCLUSION:These findings suggest that PMS2 may provide a potential reliable biomarker for CRC classification by combined immunohistochemical and mRNA analysis.

Colorectal cancer (CRC) is regarded as the third most common cancer worldwide. Although Regorafenib as a receptor tyrosine kinase inhibitor (RTKI) disrupts tumor growth and angiogenesis in metastatic CRC (mCRC) patients, drug resistance leads to poor prognosis and survival. Integrin-β1 overexpression has been proposed to be the major player in this regard. Herein, the Regorafenib-resistant human colon cancer cell line (SW-48) was induced, and the Integrin-β1 gene expression, as well as apoptosis, was assessed through the combination of small interfering RNA (siRNA) targeting Integrin-β1 and Regorafenib/Dimethyldioctadecylammonium bromide (DDAB)-methoxy poly (ethylene glycol) (mPEG)-poly-ε-caprolactone (PCL) hybrid nanoparticles (HNPs). In the current study, Regorafenib-resistant SW-48 cell line was generated in which the Regorafenib half-maximal inhibitory concentration (IC50) for non-resistant and resistant cells was 13.5±1.5 µM and 55.1±0.8 µM, respectively. The results of DLS also demonstrated that the size and the charge of the HNPs were equal to 66.56±0.5 nm and +29.5±1.2 mv, respectively. In addition, the Integrin-β1 gene expression was significantly higher in resistant cells than in non-resistant ones (P<0.05). The siRNA/HNP complexes in combination with Regorafenib/HNPs were accordingly identified as the most effective treatment to decrease the Integrin-β1 gene expression and to enhance the apoptosis rate in resistant cells (P<0.001). Overall, the study indicated that combination therapy using siRNA/HNP and Regorafenib/HNPs complex could down-regulate the Integrin-β1 gene expression and consequently trigger apoptosis, and this may potentially induce drug sensitivity.