Cell Journal (Yakhteh)
Volume:25 Issue: 2, Feb 2023

Some cationic anti-microbial peptides show a wide range of cytotoxic action versus malignant cells,which may lead to developing a novel group of antitumor medications. In the present study, the anticancer activity ofpleurocidin-like peptide WF3 isoform X2 (AMP-WF3), from the Poecilia Mexicana fish, against leukemic cell line Jurkatwas evaluated, and the cytotoxicity compared with the effects on normal cells, including peripheral blood mononuclearcells (PBMCs) and human dermal fibroblast (HDF) cells. In this experimental study, cells were treated with various dosages of AMP-WF3 for 24 hours.Using methyl thiazole tetrazolium salt reduction (MTT test), the effects of the AMP-WF3 on cell viability and toxicitywere evaluated. The impact of this peptide on apoptotic pathways was examined using flow cytometry and AnnexinV-PI stains. Additionally, the relative expression of the P53, P21, and BCL-2 genes was evaluated using a real-timepolymerase chain reaction. The Jurkat cell line was more susceptible to AMP-WF3 cytotoxicity [half-maximal inhibitory concentration(IC50)=50 μM], while normal cells (PBMCs and HDF) were less susceptible. Flow cytometry verified that the apoptoticactivity of AMP-WF3 on Jurkat cells was significantly higher than that of HDF and PBMCs. Peptide-treated Jurkat cellswere associated with increased expression of P21, and P53 genes. In contrast, the changes in P21, P53, and BCL-2genes differed in PBMCs and HDF cells. In HDF cells, simultaneous increase of P21, P53, and BCL-2, and in PBMCs,only the increase of BCL-2 was observed. Our research showed that AMP-WF3 could be developed as a novel treatment agent with minimum sideeffects for ALL patients.

Minimal residual disease (MRD) is considered the greatest prognostic factor in acute lymphoblastic leukemia(ALL). MRD is a valuable tool for anticipating impending relapse and treatment response assessment. The objective ofthe present study was to investigate whether the detection of IgH gene rearrangement using polymerase chain reaction(PCR)-based GeneScan analysis could be a complementary method to monitor MRD along with the quantitative realtimePCR (qPCR). In this cross-sectional study, we valued the MRD levels, based on the GeneScanning analysis(GSA), and then compared the data with quantitative real-time polymerase chain reaction at different time points inperipheral blood (PB) samples of adult B-lineage ALL patients (n=35). The specific polymerase chain reaction (PCR)primers for IGH gene FR-1 and fluorescence-labeled J-primer were used and analyzed by capillary gel electrophoresison a sequencer. The results of this study were compared with the previously reported MRD results obtained by the IGHrearrangements allele-specific oligonucleotide (ASO) -qPCR methods. The total concordance rate was 86.7%, with a P<0.001. MRD results obtained by GSA and ASO-qPCR methodswere concordant in all diagnostic samples and samples on the 14th and 28th days of induction therapy. The results of these 2.5years’ follow-ups demonstrated a significant correlation between the two techniques (r=0.892, P<0.001). It seems that the PCR-based GeneScan analysis of IGH gene rearrangement detection may be a valuablemolecular technique to distinguish monoclonality from polyclonality. And also, it may be a precise tool to detect theresidual leukemic DNA in the PB follow-up samples of patients.

Natural killer (NK) cells are critical immune cells for acute myeloid leukemia (AML) targeting. However,little is known about the relationship between using checkpoint inhibitors and heat shock protein 70 (Hsp70) as NK cellactivators to control AML. Therefore, the study aims to find the best formulation of Hsp70, human PD-1 (Programmedcell death protein 1) blocker, and interleukin 15 (IL-15) to activate NK cells against AML. In this experimental study, the NK cells were isolated from mononuclear cells (MNCs) byusing magnetic activation cell sorting (MACS) and were activated using the different combinations of Hsp70, PD-1blocker, and IL-15 and then followed by immunophenotyping, functional assays to estimate their killing potential, andevaluation of expression pattern of PRF1, PIK3CB, PD-1, AKT-1, FAS-L, TRAIL, and GER A and B. The expression of PD-1 was significantly (P<0.05) reduced after NK cell activation by the different formulas ofIL-15, Hsp70, and PD-1 blocker. The expression of NKG2A in the treated NK cells was reduced particularly in the IL-15(P<0.01) and IL-15+PD-1 blocker (P<0.05) groups. The addition of Hsp70 increased its expression. The cytotoxic effectof NK cells increased in all groups, especially in IL-15+PD-1 blocker besides increasing interferon-gamma (IFN-γ),Granzymes, and perforin expression (P<0.05). All IL-15+PD-1 blocker group changes were associated with the upregulationof PIK3CB and AKT-1 as key factors of NK cell activation. The presence of Hsp70 reduced IFN-γ releasing,and down-regulation of PIK3CB, AKT-1, Granzymes, and Perforin (P<0.05). We suggested the combination of IL-15 and PD-1 blocker could enhance the killing potential of AMLNKcells. Moreover, Hsp70 in combination with IL-15 and PD-1 blocker interferes activation of AML-NK cells throughunknown mechanisms.

Parkinson’s disease (PD) is a severely debilitating disease for which no suitable treatment has been foundso far. In recent years, nanoparticles (NPs) have shown therapeutic potential in PD. Thus, in the current research, forthe first time, we investigated the effects of vitamin E and TiO2 nanoparticles (TiO2-NPs) on a rat model of PD. In this experimental study, after preparation and induction of PD, rats were administratedwith vitamin E and TiO2-NPs. One day after receiving the last dose of treatments, rats were killed and substantianigra was extracted from the brain and its cell suspension was prepared. In each group, female rats were mated, andafter confirmation that the female rats were pregnant by vaginal smear test, the fetus was removed. Cell viability wasstudied by the MTT method and apoptosis, and necrosis were studied by the flow cytometry technique. Also, after RNAextraction and cDNA synthesis, the expression of Bcl-2 and circRNA 0001518 genes were studied using the real timepolymerase chain reaction (RT-PCR) technique. For analyzing the data, two-way ANOVA was used. The results showed a sharp decrease in cell viability in female PD rats and fetuses resulting from PD femalerats. Vitamin E treatment showed the greatest effect on increasing cell viability. Decreased expression of the Bcl-2 geneand increased expression of circRNA 0001518 were observed in PD conditions. Administration of vitamin E may be a good option for reducing PD-induced cell death.

The function of Th17 cells in the neuroinflammatory process in multiple sclerosis (MS) has been previouslyclarified. It has been suggested that Quercetin can influence MS due to a variety of anti-inflammatory effects. The presentstudy aimed to examine in vitro immunomodulatory aspects of Quercetin Penta Acetate as a modified compound onTh17 cells of MS patients and also to compare its effects with Quercetin. In this experimental study, peripheral blood mononuclear cell (PBMCs) were isolated andstained with CFSE then, half-maximal inhibitory concentration (IC50) values were determined using different dosesand times for Quercetin Penta Acetate, and Methyl Prednisolone Acetate. Th17 cell proliferation was analyzed by flowcytometry and the expression levels of IL-17 and RORc genes were assessed by real-time polymerase chain reaction(PCR) method. The results showed that IL-17A gene expression was inhibited by Quercetin Penta Acetate (P=0.0081), butQuercetin Penta Acetate did not have a significant inhibitory effect on Th17 cells proliferation (P= 0.59) and RORc geneexpression (P=0.1), compared to Quercetin. Taken together, our results showed some immunomodulatory aspects of Quercetin Penta Acetate onTh17 cells are more effective than Quercetin and it could be considered in the treatment of MS.

Chemotherapeutic drug resistance is the main obstacle that affects the efficacy of current therapies ofhepatocellular carcinoma (HCC), which needs to be addressed urgently. High expression of histone methyltransferaseG9a was reported to play a pivotal role in the progression of HCC. Regulatory mechanism of aberrant activation of G9ain HCC and the association with subsequent cisplatin (DDP) resistance still remains ambiguous. This study strived toinvestigate mechanism of G9a overexpression and its impact on cisplatin resistance in HCC cells. In this experimental study, we investigated effects of different concentrations of cisplatin incombination with BIX-01294 or PR-619 on viability and apoptosis of HuH7 and SNU387 cells via CCK-8 kit and flowcytometric analysis, respectively. Colony formation capacity was applied to evaluate effect of cisplatin with or withoutBIX-01294 on cell proliferation, and western blotting was used to verify expression level of the related proteins. GlobalmRNA expression profile analysis was adopted to identify differentially expressed genes associated with overexpressionof G9a. We observed that overexpression of G9a admittedly promoted cisplatin resistance in HCC cells. GlobalmRNA expression profile analysis after G9a inhibition showed that DNA repair and cell cycle progression were downregulated.Moreover, we identified that deubiquitination enzymes (DUBs) stabilized high expression of G9a in HCCthrough deubiquitination. Additionally, cisplatin could significantly inhibit proliferation of DUBs-deficient HCC cells, whilepromoting their apoptosis. Collectively, our data indicated that DUBs stabilize G9a through deubiquitination, thereby participating inthe cisplatin resistance of HCC cells. The elucidation of this mechanism contributes to propose a potential alternativeintervention strategy for the treatment of HCC patients harboring high G9a levels.

Increasing research has been focused on the development of various nanocomplexes as targeted contrastmedia in diagnostic modalities, mainly in computed tomography (CT) scan imaging. Herein, we report a new methodthat uses Triptorelin [a luteinizing hormone-releasing hormone (LHRH) agonist]-targeted gold nanoparticles (AuNPs)via alginate for early detection of cancer by molecular CT imaging. In the experimental study, the formed multifunctional AuNPs coated with alginate conjugatedwith Triptorelin peptide (Triptorelin-Alginate-AuNPs) were synthesized and characterized via different techniques,including transmission electron microscopy (TEM), dynamic light scattering (DLS), and fourier transform infrared (FTIR)spectroscopy. The MTT assay was applied to calculate the toxicity of the NPs. The results indicated that the formed Triptorelin-Alginate-AuNPs with an Au core size of ~18 nm arenoncytotoxic at 127-, 254-, 381- and 508-mM concentrations and revealed significant improvement in the attenuationof X-rays intensity and contrast to noise ratio (CNR), compared with non-targeted cells at the highest energies (90, 120,140 kVp). At 90 kVp, compared to non-targeted cells, targeted cells (Triptorelin-Alginate-AuNPs) enable 1.58, 1.69, 3.7and 3.43 times greater contrast at a concentration of 127 mM, 254 mM, 381 mM, and 508 mM, respectively. These results suggest that the developed Triptorelin-Alginate-AuNPs may be considered an effectivecontrast agent for molecular CT imaging of gonadotropin-releasing hormone (GnRH) receptor-expressing cancer cells.

Satellite cells play an important role in muscle regeneration, which this process can be affected by differentgenes including PAX7 and MyoD. Exercise training known as an important strategy for mediating the satellite cell’sfunction. Therefore, the main purpose of the present study is to investigate the changes in PAX7 and MyoD proteinexpression in response to eccentric and concentric resistance exercise in healthy young men. In this semi-experimental and cross-sectional study, 10 healthy men (age range 18-30 yearsold) participated. They were randomly divided into two equal groups (n=5) to perform one of two high-intensity eccentricor concentric knee extensions muscle contraction protocols. The contractions included a maximum of 12 sets of 10repetitions, with a 30 second rest time interval between sets. PAX7 and MyoD protein expression was assessed usingImmunohistochemistry analysis from the Vastus Lateralis muscle needle biopsy samples that have been taken 24hours before and 3 to 4 hours after the end of the exercise protocol. We observed that the PAX7 protein expression level increased significantly after eccentric (47.75%) andconcentric (39.21%) (P=0.01) intervention. While, the MyoD protein expression level reduced (38.14%) significantlyfollowing acute eccentric resistance exercise (P=0.01). It seems that eccentric or concentric muscular contraction modulates the expression of PAX7 and MyoDprotein expression in the skeletal muscle, with further effects observed in eccentric resistance exercise.