Cell Journal (Yakhteh)

85-91

Authors: Sepideh Shahkarami; Samareh Younesian; Shahrbano Rostami; Farzad Kompani; Davood Bashash; Seyed Asadollah Mousavi; Seyed H. Ghaffari

Objective: 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 of the 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 realtime PCR (qPCR). Materials and Methods: 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 in peripheral 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 electrophoresis on a sequencer. The results of this study were compared with the previously reported MRD results obtained by the IGH rearrangements allele-specific oligonucleotide (ASO) -qPCR methods. Results: The total concordance rate was 86.7%, with a P<0.001. MRD results obtained by GSA and ASO-qPCR methods were concordant in all diagnostic samples and samples on the 14th and 28th days of induction therapy. The results of these 2.5 years’ follow-ups demonstrated a significant correlation between the two techniques (r=0.892, P<0.001). Conclusion: It seems that the PCR-based GeneScan analysis of IGH gene rearrangement detection may be a valuable molecular technique to distinguish monoclonality from polyclonality. And also, it may be a precise tool to detect the residual leukemic DNA in the PB follow-up samples of patients.

102-109

Authors: Behdokht Jamali; Malihe Entezari; Nahid Babaei; Mehrdad Hashemi

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

118-125

Authors: Junhao Fu; Min Yu; Wenxia Xu; Shian Yu

Objective: Chemotherapeutic drug resistance is the main obstacle that affects the efficacy of current therapies of hepatocellular carcinoma (HCC), which needs to be addressed urgently. High expression of histone methyltransferase G9a was reported to play a pivotal role in the progression of HCC. Regulatory mechanism of aberrant activation of G9a in HCC and the association with subsequent cisplatin (DDP) resistance still remains ambiguous. This study strived to investigate mechanism of G9a overexpression and its impact on cisplatin resistance in HCC cells. Materials and Methods: In this experimental study, we investigated effects of different concentrations of cisplatin in combination with BIX-01294 or PR-619 on viability and apoptosis of HuH7 and SNU387 cells via CCK-8 kit and flow cytometric analysis, respectively. Colony formation capacity was applied to evaluate effect of cisplatin with or without BIX-01294 on cell proliferation, and western blotting was used to verify expression level of the related proteins. Global mRNA expression profile analysis was adopted to identify differentially expressed genes associated with overexpression of G9a. Results: We observed that overexpression of G9a admittedly promoted cisplatin resistance in HCC cells. Global mRNA 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 HCC through deubiquitination. Additionally, cisplatin could significantly inhibit proliferation of DUBs-deficient HCC cells, while promoting their apoptosis. Conclusion: Collectively, our data indicated that DUBs stabilize G9a through deubiquitination, thereby participating in the cisplatin resistance of HCC cells. The elucidation of this mechanism contributes to propose a potential alternative intervention strategy for the treatment of HCC patients harboring high G9a levels.

135-142

Authors: Somayeh Karimi Majd; Mandana Gholami; Behzad Bazgir

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