فهرست مطالب

Molecular Biology Research Communications - Volume:8 Issue:2, 2019
  • Volume:8 Issue:2, 2019
  • تاریخ انتشار: 1398/03/11
  • تعداد عناوین: 5
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  • Ahmad Jalilvand, Mohammad Soleiman Soltanpour * Pages 53-58
    Colorectal cancer (CRC) is one of the common causes of cancer death in Iranian population. Both genetic and epigenetic changes have been implicated in CRC pathogenesis. DACT2 gene as one of the WNT signaling pathway inhibitor was shown to display tumor suppressor activity in many cancers. The aim of present study was to investigate the methylation status of DACT2 gene and its association with methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism in CRC patients. Fifty formalin-fixed paraffin-embedded cancerous and adjacent healthy tissues obtained from CRC patientwere investigated. Genomic DNA was isolated using a FFPE commercial DNA extraction kit. The methylation status was evaluated by methylation specific PCR. Genotyping of MTHFR C677T polymorphism was performed using PCR-RFLP technique. Statistical analysis was done by GraphPad Prism 8. Results indicated that the frequency of methylated DACT2 gene was significantly higher in cancerous tissue relative to adjacent healthy tissue (P<0.001). DACT2 gene methylation was significantly more common among carriers of MTHFR 677CC genotype (P=0.035) and significantly less common among carriers of MTHFR 677T allele (P value =0.006). In conclusion the present study identified DACT2 gene methylation as a significant risk factor for CRC development. Moreover, the low frequency of DACT2 gene methylation among carriers of MTHFR 677T allele may confer a protective role for this common polymorphism against CRC risk.
    Keywords: Methylation, Colorectal cancer, DACT2, Methylenetetrahydrofolate reductase
  • Hongzhi Bai, Yanhua Zhang, Haiqiu Yu, Muhammad Irfan, Huang Yuqian, Han Mei, Jinfeng Yang, Ning Liu, Hui Wang *, Xiaori Han Pages 59-68
    Arachis hypogaea. L is a legume of economic importance, which is nodulated by Bradyrhizobium, a slow-growing bacteria. However there is no well characterization of this rhizobia in many areas of China. In the present study, cross-inoculation experiments were performed in cowpea and soybean. The isolated bacteria strains were characterized physiologyically, biochemically and identified through 16S rDNA sequence analysis showing that it belongs to Bradyrhizobium japonicum. The genetic diversity of the seventeen isolated strains were assessed through PCR-RFLP of 16S rDNA and 16S-23S rDNA IGS region. Cross inoculation test indicated that isolates could nodulate cowpea but not soybean. The cluster analysis based on physiological and biochemical characteristics showed the lower correlation between isolates and sites. The isolates were grouped into four clusters based on 16S rDNA gene sequence analysis. Thirteen polymorphisms were variable across all observations in 16S rDNA RFLP and six different IGS types from isolates. The results implies that there was some association between geographical factor and phylogenetic diversity of indigenous Bradyrhizobium isolates.
    Keywords: Peanut, Bradyrhizobium, Phylogenetic diversity, Cross inoculation
  • Nammodn Sahar, Shakila Bibi, Nosheen Masood *, Rani Faryal Pages 69-77
    Asthma is a disease marked by inflammation of airways with an increasing incidence rate worldwide especially among Asian population. Spleen tyrosine kinase (Syk) is known to be involved in regulation of such inflammation response and thereby rendering its inevitable importance among asthma patients. DNA extraction followed by PCR and sequencing was performed for genomic analysis, mRNA analysis was done by RT PCR whereas Western blot and ELISA was used for protein study. Image J and UNAFOLD were also used for Bioinformatics analysis.The mean age of patients and controls were 31.1±9.3 and 30.4±6.1 years respectively. Results of sequencing showed nonsense exonic mutations in exon 3 at g.25710G>A and g.25722G>A positions. Substitution mutations in introns were also found at g.25827G>A (intron 3), g.63425C>T (intron 8) and g.63445T>G (intron 8). Significantly increased levels of IgE and significantly decreased expression of Syk at transcriptional level was found in patients compared to controls. The western blot results of asthmatic samples and healthy controls revealed that Syk has comparatively low expression in diseased individual’s PBMCs.SYK has been found to be altered in DNA, mRNA and protein expression in asthma patients among Pakistani population therefore patients should be treated according to their Syk status for more effective recovery.
    Keywords: Spleen tyrosine kinase, Expression, SSCP, ELISA
  • Samaneh Tokhanbigli, Kaveh Baghaei *, Ali Asadirad, Seyed Mahmoud Hashemi, Hamid Asadzadeh, Aghdaei, Mohammad Reza Zali Pages 79-89
    Mesenchymal stem cells (MSCs) are well known due to their immunomodulatory effect, but the exact mechanisms have not been defined. Several studies demonstrated that the exerted immunoregulatory effect of these cells could be mediated by paracrine factors to illustrate, cytokines, chemokine, and among which, extracellular vesicles are one of them to play a crucial role. Moreover, it is assumed that extracellular vesicles are an essential player in intracellular communication by transferring their component. In this respect, the efficiency of conditioned media and exosomes was compared to illustrate a practical approach to cell-free based therapies. In the current study, we investigated the effect of both MSCs conditioned media (MSC-CM) and MSCs-derived exosomes on the expression of pro-inflammatory and anti-inflammatory cytokines in peripheral blood mononuclear cells (PBMCs). In this regard, isolated PBMCs were treated with MSC-CM and MSC-derived exosome as separated groups. Expression of inflammatory and anti-inflammatory markers was evaluated by Real-time PCR and ELISA. The immunoregulatory effect of MSC-CM on pro-inflammatory and anti-inflammatory genes, such as IL-12b, iNOS, EGR-2, IL-10 with an exception in case of IL-6 was more significant. Whereas in protein levels IL-10 showed the most substantial difference in exosome treated groups. It could be assumed that MSC-CM has more immunoregulatory impact on monocyte in contrast with exosomes.Taken together, by considering the recent approaches to cell-free therapy and the immunoregulatory impact of MSCs, yet relatively little is known about the efficacy of human-MSC-CM and secreted exosome compared with each other.
    Keywords: Mesenchymal stromal cells, Conditioned media, Exosomes, Pro-inflammatory, anti-inflammatory cytokines
  • Parisa Rabiei, Hassan Mohabatkar *, Mandana Behbahani Pages 91-98
    A great number of researches over the last years are allocated to know cancer reasons, prevention and treatment strategies. Bacterial infections are one of the promoting factors in cancer development. The present study was carried out to study effects of heat-killed bacteria on cancer cell lines MCF7 and HT-29. To this purpose, four bacterial strains including Salmonella typhi, Staphylococcus epidermidis, Escherichia coli and Pseudomonas aeruginosa were assayed. Thermal inactivation method was used to kill the bacteria and preserve the bacterial surface proteins unchangeable. The concentrations of 0.01, 0.1, 0.5 and 1 mg/ml of inactivated bacteria were prepared to evaluate the effects of heat-inactivated bacterial solutions on MCF7 and HT-29 cell lines. MTT assay was used to measure the cell viability of cancer cells treated with different concentration of inactivated bacterial solutions.The MTT assay results after 48 hours showed that the heat-killed bacterial solutions were able to induce the proliferation of both cancer cell lines. In addition, the most cell viability in MCF-7 cell line was seen in samples treated with S. epidermidis, while in HT29 cells, the most one was seen in S. typhi treated samples. It was concluded that bacterial infections are cancer-deteriorating agents, and any species of bacteria is specific to certain cancerous tissue.
    Keywords: Inflammatory responses, Chronic infections, MTT-assay, Heat inactivation, Colon cancer, Breast cancer