MiR-21-5p knockdown inhibits epithelial to mesenchymal transition in A549 lung adenocarcinoma cells by upregulating RhoB.

BACKGROUND: MiR-21-5p is a highly expressed microRNA that plays an important role in various cancer-promoting processes, including anchorage-independent growth, invasion, migration metastasis, and drug resistance in lung cancer. Studies indicate that miR-21-5p may contribute to these processes by promoting epithelial-mesenchymal transition (EMT). Ras homolog gene family member B (RhoB), a gene downregulated by miR-21-5p, has also been linked to EMT in lung cancer. However, the role of the miR-21-5p/RhoB axis in EMT regulation in lung adenocarcinoma remains unclear. In this study, we aimed to investigate the regulatory role of the miR-21-5p/RhoB axis in EMT and related in vitro functional characteristics such as migration, invasion, cisplatin resistance, and the formation of tumor spheroids. METHODS AND RESULTS: A549 cells were transfected with the miR-21-5p inhibitor, RhoB siRNA, and their corresponding negative controls. Wound healing, transwell invasion, Methyl thiazole tetrazolium (MTT), and sphere formation assays were also performed to evaluate the migration, invasion, cisplatin resistance, and anchorage-independent growth of A549 cells. RT-qPCR was used to determine the mRNA expression levels of EMT markers. MiR-21-5p knockdown inhibited migration, invasion, cisplatin resistance, and sphere formation while upregulating E-cadherin and downregulating Slug. Furthermore, RhoB silencing restored EMT and related in vitro functional characteristics in A549 cells. CONCLUSIONS: Knockdown of miR-21-5p inhibits EMT and related in vitro functional characteristics by upregulating RhoB, suggesting that miR-21-5p may promote EMT through downregulation of RhoB.

Diatomite-Based, Flexible SERS Immunosensor Platform for Rapid, Specific, and Sensitive Detection of Circulating Cancer-Specific Protein Biomarkers in Serum Using Raman Probes.

Cancer is one of the most actively researched diseases having a high mortality rate when not detected at an early stage. Thus, rapid, simultaneous, and sensitive quantification of cancer biomarkers plays an important role in early diagnosis, with patient impact to disability adjusted life years. Herein, a diatomite-based SERS flexible platform for the rapid and sensitive detection of circulating cancer-specific protein biomarkers in serum is presented. In this approach, diatomite/AgNPs strips with maximum SERS activity prepared using the layer-by-layer (LbL) technique were modified with specific antibodies, and specific antigens (HER2, CA15-3, PSA, and MUC4) were captured and detected. By using Raman probes specific to the captured antigens in serum, a SERS limit of detection (LOD) of 0.1 ng/mL was measured (calculated LOD < 0.1 ng/mL). This value is lower than the cutoff amount of cancer antigens in the person's blood. The specificity for the antigens of each antibody was calculated to be higher than 95%. As a result, an immunosensor for rapid detection of cancer biomarkers in serum with good specificity, high sensitivity, good reproducibility, and low cost has been demonstrated. Overall, we show that the prepared diatomite-based SERS substrate with a high surface-to-volume ratio is a useable platform for immunoassay tests.

Effect of different molecular weight and terminal group PLGA on docetaxel nanoparticles: characterization and cytotoxic activity of castration-resistant prostate cancer cells.

The choice of polymer and its compatibility with drug used determine the fate of nanoparticle in therapy. There has been limited sources about effect of resomer differentiation in nanoparticle related with physical and chemical properties and also biological activities of product. Therefore, we aimed to formulate docetaxel-loaded polylactic-co-glycolic acid nanoparticles with different molecular weights (Resomer 502 and 504) and terminal groups (Resomer 502H and 504H) and to investigate the effect of these resomers on nanoparticle character, prostate cancer, and healthy cells. Docetaxel-loaded PLGA nanoparticles were prepared by single emulsion solvent evaporation method. Surface characterizations were carried out by zeta sizer and scanning electron microscopy. Encapsulation efficiency, in vitro drug release profiles, and cytotoxic activity were determined. Main effect on the surface morphology of nanoparticles was the molecular weight of the polymer. The groups with acid terminal function have higher encapsulation and reaction efficiency. In all formulations, in vitro release was observed after 334 h at pH 7.4 and 240 h at pH 5.6. Also, the groups with high molecular weight showed selective cytotoxicity. These resomers especially RG 504 and RG 504H have potential to be used as a low-dose and high-efficiency extended-release drug delivery system in the treatment of prostate cancer.

Anticancer Activity Study and Density Functional/Time-Dependent Density Functional Theory (DFT/TD-DFT) Calculations of 2(3),9(10),16(17),23(24)-Tetrakis-(6-Methylpyridin-2-Yloxy)Phthalocyaninato Zn(II).

Photodynamic therapy (PDT) is one of the major therapeutic methods for the treatment of infectious diseases and cancer. Recently, cell culture has been used to determine the effect of a given substance on various pathological conditions, such as cancer. In this study, we aimed to investigate the effect of a Zn- phthalocyanine (ZnPc) derivative on selected cancer cells via a cell culture medium. Methylthiazole tetrazolium (MTT) assay was applied to evaluate the cytotoxic activity of 2(3),9(10),16(17),23(24)-tetrakis-(6-methylpyridin-2-yloxy)phthalocyaninato Zn(II) on rat glioma cells (C6 glioma), human lung cancer cells (H1299) and human umbilical vein endothelial cells (HUVEC). The levels of the lipid peroxidation were determined by measuring the amount of the thiobarbituric acid reactive substance (TBARS) produced using the TBARS assay. The relationship between the oxidative damage and the effective concentration of cytotoxic ZnPc was determined from the results. The apoptotic and genotoxic effects of the phthalocyanine (Pc) were also investigated. Density functional/time-dependent density functional theory (DFT/TD-DFT) methods were used to determine the molecular excited state properties of the ZnPc and chloroaluminum phthalocyanine (ClAlPc) previously reported by Castilho-Fernandes et al. The computed and experimental data were used to establish a link between the electronic and anticancer properties of the Pcs.

Antitumoral effect of a selective Rho-kinase inhibitor Y-27632 against Ehrlich ascites carcinoma in mice.

BACKGROUND: The Rho proteins and Rho-kinase (ROCK) enzymes are responsible for signal transduction, and cause cell permeability, contractility, differentiation, migration, proliferation or apoptosis depending on cell types. All of these functions are vital for cancer initiation and progression. In this study, the preventive and protective effects of a selective ROCK inhibitor Y-27632 against Ehrlich ascites carcinoma in Swiss albino mice were investigated. METHODS: Adult male albino mice were divided into five equal groups, and Y-27632 (0.1, 1, and 10 mg/kg) was given to groups as two steps; before (pre-carcinoma) and after inoculation of carcinoma cell suspensions (post-carcinoma). At the end of the experiments (at day 15), cardiac blood samples, the ascitic fluid, and intestinal specimens were collected for histopathology and biochemical investigation. RESULTS: Significant decreases in the body weight and immunostaining scores in small and large intestine for ROCK2, preservation of serum glutathione (GSH) levels, and an increase in tumor level of nitric oxide were recorded in groups pretreated with Y-27632. However, treatment with Y-27632 after tumor inoculation did not affect body weight and ROCK2 immunostaining scores, increased serum MDA levels, and decreased GSH levels. CONCLUSIONS: This is the first study on the effectiveness of Y-27632 in this experimental tumor model. Our findings provided direct evidence for ROCK involvement in tumor development. These data suggest that pretreatment with Y-27632 has a protective effect against tumor formation.

CDC 25A gene 263C/T, -350C/T, and -51C/G polymorphisms in breast carcinoma.

The family of cell division cycle 25 (CDC25) phosphatase is one of the important regulators of the cell cycle progression. In mammalian cells, three isoforms have been identified: CDC25A, CDC25B, and CDC25C. CDC25A is required to enter S time, and the overexpression of this phosphatase accelerates the entrance to S time. CDC25A overexpression could render tumor cells less sensitive to DNA replication checkpoints, thereby contributing to their genomic instability. We aimed to investigate, for the first time, the frequency of human CDC25A gene SNPs in metastatic and non-metastatic breast cancer. Total number of 281 eligible patients with histologically confirmed incident of breast cancer and 137 cancer-free controls were included. The detection of CDC25A gene polymorphisms was achieved with real-time polymerase chain reaction and restriction fragment length polymorphism techniques. We found that the 263C/T polymorphism was significantly associated with breast cancer and risk of metastasis. The -350C/T polymorphism in the promoter region of CDC25A gene was found to associate with neither breast cancer nor metastasis. The other promoter polymorphism -51C/G in the CDC25A gene associated with breast cancer but not associated with metastasis. These data suggest that 263C/T and -51C/G polymorphisms of CDC25A gene could be candidate markers for earlier diagnosis and targets for breast cancer therapy.

[Evaluation of the patients with low levels of anti-HCV positivity]. / Düsük titrede anti-HCV pozitifligi tespit edilen hastalarin irdelenmesi.

Laboratory diagnosis of hepatitis C virus (HCV) infection is based on the detection of anti-HCV antibodies by enzyme immunoassay (EIA) or chemiluminescence immunoassay (CIA) techniques. However, a consensus related to the problem of low titer (Serum/Cut-off; S/C= 1.0) anti-HCV antibodies is still lacking. This study was aimed to evaluate the clinical status of the patients with low titer anti-HCV antibodies detected by third generation anti-HCV tests during january 2007-December 2007. Two hundred and fifteen sera with anti-HCV S/C values between 1-5, detected by a commercial test system (Vitros EC Immunodiagnostic System, 3rd generation anti-HCV test, Ortho-Clinical Diagnostics, USA) with a sensitivity of 100% and specificity of 99.7%, as indicated by the supplier, were included to the study. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were determined by using chemiluminescence assay (Roche Diagnostics, Germany) and HCV-RNA was detected by real-time PCR (Flurion HCV QNP 2.1). Hundred and thirty six (63.3%) of the patients were female and 79 (36.7%) were male. The mean age of the patients was 50.2 +/- 18.9 years. In 18 (8.3%) patients ALT and/or AST levels were high and two of them were infected with hepatitis A while the remaining two with hepatitis B virus. HCV-RNA positivity (15.6 x 10(6); 4.3 x 10(5) and 2.6 x 10(3) IU/ml, respectively) was detected in three patients (1.4%) with S/C values of 3.69, 4.46 and 4.59, respectively. These three patients were older than 50 years, had high ALT levels and were chronic renal failure patients undergoing dialysis for at least one year. It was observed that after 4-6 weeks anti-HCV titers increased (S/C values were 15.1, 6.5 and 11.8, respectively) in the serum samples of these patients. The data obtained from this study emphasizes the problem of low titer positive anti-HCV results. It could be concluded that in case of low titer anti-HCV values, the result should be confirmed by RIBA, although its use is a matter of debate due to its low sensitivity, and HCV-RNA tests. Based on these data it seemed that changing the anti-HCV S/C ratio would not be a solution for the problem of low titer anti-HCV positive results.