• Title/Summary/Keyword: molecular profile

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Comparative proteome analysis of diploid and tetraploid root in Platycodon grandiflorum

  • Kwon, Soo Jeong;Roy, Swapan Kumar;Yoo, Jang-Hawan;Cho, Seong-Woo;Kim, Hag Hyun;Boo, Hee Ock;Woo, Sun-Hee
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.123-123
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    • 2017
  • In spite of the potential medicinal significance and a wide range of pharmacologic properties of Platycodon grandiflorum, the molecular mechanism of its roots is still unknown. The present study was conducted to profile proteins from 3, 4 and 5 months aged diploid and tetraploid roots of Platycodon grandiflorum using high throughput proteome approach. Two-dimensional gels stained with CBB, a total of 68 differential expressed proteins were identified from the diploid root out of 767 protein spots using image analysis by Progenesis SameSpot software. Out of total differential expressed spots, 29 differential expressed protein spots (${\geq}2-fold$) were analyzed using LTQ-FTICR MS whereas a total of 24 protein spots were up-regulated and 5 protein spots were down-regulated. On the contrary, in the case of tetraploid root, a total of 86 differential expressed proteins were identified from tetraploid root out of 1033 protein spots of which a total of 39 differential expressed protein spots (${\geq}2-fold$) were analyzed using LTQ-FTICR MS whereas a total of 21 protein spots were up-regulated and a total of 18 protein spots were down-regulated. It was revealed that the identified proteins from the explants were mainly associated with the nucleotide binding, oxidoreductase activity, transferase activity. Taken together, the identified proteins may be helpful to identify key candidate proteins for genetic improvement of plants.

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Toxicogenomics Study on TK6 Human Lymphoblast Cells Treated with Mitomycin C

  • Kim, Joo-Hwan;Koo, Ye-Mo;Lee, Woo-Sun;Suh, Soo-Kyung;Kang, Jin-Seok;Han, Eui-Sik;Kim, Seung-Hee;Park, Sue-N.
    • Molecular & Cellular Toxicology
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    • v.3 no.3
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    • pp.165-171
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    • 2007
  • Mitomycin C (MMC), an antitumor antibiotic isolated from Streptomyces caespitosus, is used in chemotherapy of gastric, bladder and colorectal cancer. MMC is activated in vivo to alkylate and crosslink DNA, via G-G interstrand bonds, thereby inhibiting DNA synthesis and transcription. This study investigates gene expression changes in response to MMC treatment in order to elucidate the mechanisms of MMC-induced toxicity. MMC was admistered with single dose (0.32 and 1.6 ${\mu}M$) to TK6 cells. Applied Biosystem's DNA chips were used for identifying the gene expression profile by MMC-induced toxicity. We identified up- or down-regulated 90 genes including cyclin M2, cyclin-dependent kinase inhibitor 1A (p21, cip1), programmed cell death 1, tumor necrosis factor (ligand) superfamily, member 9, et al. The regulated genes by MMC associated with the biological pathways apoptosis signaling pathway. Further characterization of these candidate markers related to the toxicity will be useful to understand the detailed mechanism of action of MMC.

Isolation of formaldehyde-responsive proteins in Arabidopsis (Formaldehyde에 반응하는 애기장대 단백질의 분리)

  • Kwon, Mi;Park, Hyun Jin;Seo, Jae Hyun
    • Journal of the Korean Wood Science and Technology
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    • v.35 no.4
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    • pp.52-60
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    • 2007
  • Plant can detoxify the effect of the volatile organic compounds (VOC) such as formaldehyde and toluene, however, mechanisms of VOC detoxification are largely unknown in plant system. This study was performed to investigate phenotypic changes of Arabidopsis seedlings upon treatment of either formalin or toluene. Formalin treatment up to twenty four hours didn't cause any significant phenotypic damages on the leaf surface of 27 DAG Arabidopsis seedlings. However, the protein profile of formalin-treated seedlings was significantly different from that of mock control. Using automated electrophoresis system, the molecular weight of each formaldehyde-responsive protein (FRP) was predicted and its formaldehyde-dependent expression was confirmed at transcription level by quantitative real-time RT-PCR analysis. Four FRPs isolated in this study are the novel proteins with unknown functions but highly homologous to the stress-related proteins.

Expression Profiling of WSSV ORF 199 and Shrimp Ubiquitin Conjugating Enzyme in WSSV Infected Penaeus monodon

  • Jeena, K.;Prasad, K. Pani;Pathan, Mujahid Khan;Babu, P. Gireesh
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.8
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    • pp.1184-1189
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    • 2012
  • White spot syndrome virus (WSSV) is one of the major viral pathogens affecting shrimp aquaculture. Four proteins, WSSV199, WSSV 222, WSSV 249 and WSSV 403, from WSSV are predicted to encode a RING-H2 domain, which in presence of ubiquitin conjugating enzyme (E2) in shrimp can function as viral E3 ligase and modulate the host ubiquitin proteasome pathway. Modulation of host ubiquitin proteasome pathway by viral proteins is implicated in viral pathogenesis. In the present study, a time course expression profile analysis of WSSV Open Reading Frame (ORF) 199 and Penaeus monodon ubiquitin conjugating enzyme (PmUbc) was carried out at 0, 3, 6, 12, 24, 48 and 72 h post WSSV challenge by semi-quantitative RT-PCR as well as Real Time PCR. EF1${\alpha}$ was used as reference control to normalize the expression levels. A significant increase in PmUbc expression at 24 h post infection (h.p.i) was observed followed by a decline till 72 h.p.i. Expression of WSSV199 was observed at 24 h.p.i in WSSV infected P. monodon. Since the up-regulation of PmUbc was observed at 24 h.p.i where WSSV199 expression was detected, it can be speculated that these proteins might interact with host ubiquitination pathway for viral pathogenesis. However, further studies need to be carried out to unfold the molecular mechanism of interaction between host and virus to devise efficient control strategies for this chaos in the shrimp culture industry.

Reversal of Multidrug Resistance in Mouse Lymphoma Cells by Extracts and Flavonoids from Pistacia integerrima

  • Rauf, Abdur;Uddin, Ghias;Raza, Muslim;Ahmad, Bashir;Jehan, Noor;Siddiqui, Bina S;Molnar, Joseph;Csonka, Akos;Szabo, Diana
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.1
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    • pp.51-55
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    • 2016
  • Phytochemical investigation of Pistacia integerrima has highlighted isolation of two known compounds naringenin (1) and dihydrokaempferol (2). A crude extract and these isolated compounds were here evaluated for their effects on reversion of multidrug resistance (MDR) mediated by P-glycoprotein (P-gp). The multidrug resistance P-glycoprotein is a target for chemotherapeutic drugs from cancer cells. In the present study rhodamine-123 exclusion screening test on human mdr1 gene transfected mouse gene transfected L5178 and L5178Y mouse T-cell lymphoma cells showed excellent MDR reversing effects in a dose dependent manner. In-silico molecular docking investigations demonstrated a common binding site for Rhodamine123, and compounds naringenin and dihydrokaempferol. Our results showed that the relative docking energies estimated by docking softwares were in satisfactory correlation with the experimental activities. Preliminary interaction profile of P-gp docked complexes were also analysed in order to understand the nature of binding modes of these compounds. Our computational investigation suggested that the compounds interactions with the hydrophobic pocket of P-gp are mainly related to the inhibitory activity. Moreover this study s a platform for the discovery of novel natural compounds from herbal origin, as inhibitor molecules against the P-glycoprotein for the treatment of cancer.

Impact of Global and Gene-Specific DNA Methylation in de Novo or Relapsed Acute Myeloid Leukemia Patients Treated with Decitabine

  • Zhang, Li-Ying;Yuan, You-Qing;Zhou, Dong-Ming;Wang, Zi-Yan;Ju, Song-Guang;Sun, Yu;Li, Jun;Fu, Jin-Xiang
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.1
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    • pp.431-437
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    • 2016
  • In this investigation, global DNA methylation patterns and the specific methylation status of 5 genes were studied in DNA from peripheral blood (PB) and impact on progression free survival (PFS) and overall-survival (OS) in patients with de novo or relapsed acute myeloid leukemia (AML) treated with decitabine-based regimens waas assessed. DNA was isolated from PB samples at the time of -1, 1, and 7 days of chemotherapy. Global methylation was determined by ELISA, and the CpG island DNA methylation profile of 5 genes using a DNA methylation PCR system. Our data demonstrated that patients with a high level of 5-mC had a poor prognosis after demethylation therapy and those who have low levels of 5-mC in PB achieved higher CR and better SO, but there was no significant correlation found between the 5-mC levels and other clinical features before treatment except the disease status. Higher methylation status of Sox2 and Oct4 genes was associated with differential response to demethylation therapy. A relatively low methylation percentage in one or both of these two genes was also associated with longer OS after decitabine based chemotherapy. We also suggest that global DNA and Oct-4/Sox2 methylation might impact on the pathogenesis of leukemia and play an important role in the initiation and progression. Moreover, dynamic analysis of 5-mC and Oct-4/Sox2 in peripheral blood nucleated cells of leukemia patients may provide clues to important molecular diagnostic and prognostic targets.

Genomic Screening for Targets Regulated by Berberine in Breast Cancer Cells

  • Wen, Chun-Jie;Wu, Lan-Xiang;Fu, Li-Juan;Yu, Jing;Zhang, Yi-Wen;Zhang, Xue;Zhou, Hong-Hao
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.6089-6094
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    • 2013
  • Berberine, a common isoquinoline alkaloid, has been shown to possess anti-cancer activities. However, the underlying molecular mechanisms are still not completely understood. In the current study, we investigated the effects of berberine on cell growth, colony formation, cell cycle distribution, and whether it improved the anticancer efficiency of cisplatin and doxorubicin in human breast cancer estrogen receptor positive (ER+) MCF-7 cells and estrogen receptor negative (ER-) MDA-MB-231 cells. Notably, berberine treatment significantly inhibited cell growth and colony formation in the two cell lines, berberine in combination with cisplatin exerting synergistic growth inhibitory effects. Accompanied by decreased growth, berberine induced G1 phase arrest in MCF-7 but not MDA-MB-231 cells. To provide a more detailed understanding of the mechanisms of action of berberine, we performed genome-wide expression profiling of berberine-treated cells using cDNA microarrays. This revealed that there were 3,397 and 2,706 genes regulated by berberine in MCF-7 and MDA-MB-231 cells, respectively. Fene oncology (GO) analysis identified that many of the target genes were involved in regulation of the cell cycle, cell migration, apoptosis, and drug responses. To confirm the microarray data, qPCR analysis was conducted for 10 selected genes based on previously reported associations with breast cancer and GO analysis. In conclusion, berberine exhibits inhibitory effects on breast cancer cells proliferation, which is likely mediated by alteration of gene expression profiles.

The phytohormone abscisic acid increases triacylglycerol content in the green microalga Chlorella saccharophila (Chlorophyta)

  • Contreras-Pool, Patricia Yolanda;Peraza-Echeverria, Santy;Ku-Gonzalez, Angela Francisca;Herrera-Valencia, Virginia Aurora
    • ALGAE
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    • v.31 no.3
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    • pp.267-276
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    • 2016
  • Microalgae are currently a very promising source of biomass and triacylglycerol (TAG) for biofuels. In a previous study, we identified Chlorella saccharophila as a suitable source of oil for biodiesel production because it showed high biomass and lipid content with an appropriate fatty acid methyl esters profile. To improve the TAG accumulation in C. saccharophila, in this study we evaluated the effect of abscisic acid (ABA) addition on cell concentration, lipid content and TAG production in this microalga. First, we evaluated the effects of four ABA concentrations (1, 4, 10, and 20 μM) added at the beginning of a single-stage cultivation strategy, and found that all concentrations tested significantly increased cell concentration and TAG content in C. saccharophila. We then evaluated the addition of 1 μM ABA during the second stage of a two-stage cultivation strategy and compared it with a nitrogen deficiency treatment (ND) and a combination of ND and ABA (ND + ABA). Although ABA alone significantly increased lipid and TAG contents compared with the control, ND showed significantly higher TAG content, and ND + ABA showed the highest TAG content. When comparing the results of both strategies, we found a superior response in terms of TAG accumulation with the addition of 1 μM ABA at the beginning of a single-stage cultivation system. This strategy is a simple and effective way to improve the TAG content in C. saccharophila and probably other microalgae as a feedstock for biodiesel production.

Gene Microarray Analysis for Porcine Adipose Tissue: Comparison of Gene Expression between Chinese Xiang Pig and Large White

  • Guo, W.;Wang, S.H.;Cao, H.J.;Xu, K.;Zhang, J.;Du, Z.L.;Lu, W.;Feng, J.D.;Li, N.;Wu, C.H.;Zhang, L.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.1
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    • pp.11-18
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    • 2008
  • We created a cDNA microarray representing approximately 3,500 pig genes for functional genomic studies. The array elements were selected from 6,494 cDNA clones identified in a large-scale expressed sequence tag (EST) project. These cDNA clones came from normalized and subtracted porcine adipose tissue cDNA libraries. Sequence similarity searches of the 3,426 ESTs represented on the array using BLASTN identified 2,790 (81.4%) as putative human orthologs, with the remainder consisting of "novel" genes or highly divergent orthologs. We used the gene microarray to profile transcripts expressed by adipose tissue of fatty Chinese Xiang pig (XP) and muscley Large White (LW). Microarray analysis of RNA extracted from adipose tissue of fatty XP and muscley LW identified 81 genes that were differently expressed two fold or more. Transcriptional differences of four of these genes, adipocyte fatty acid binding protein (aP2), stearyl-CoA desaturase (SCD), sterol regulatory element binding transcription factor 1 (SREBF1) and lipoprotein lipase (LPL) were confirmed using SYBR Green quantitative RT-PCR technology. Our results showed that high expression of SCD and SREBF1 may be one of the reasons that larger fat deposits are observed in the XP. In addition, our findings also illustrate the potential power of microarrays for understanding the molecular mechanisms of porcine development, disease resistance, nutrition, fertility and production traits.

Lack of Utility of Cytokeratins in Differentiating Pseudocarcinomatous Hyperplasia of Granular Cell Tumors from Oral Squamous Cell Carcinoma

  • Al-Eryani, Kamal;Karasneh, Jumana;Sedghizadeh, Parish P;Ram, Saravanan;Sawair, Faleh
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.4
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    • pp.1785-1787
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    • 2016
  • Granular cell tumor (GCT) of the oral cavity is a benign lesion. Half of oral GCTs demonstrate pseudocarcinomatous hyperplasia (PCH) of the mucosa which can mimic invasive islands of oral squamous cell carcinoma (SCC). Such similarity can be confusing when diagnosing or evaluating the two conditions, potentially leading to misdiagnosis or misclassification. Indeed, several misdiagnosed cases of oral GCT have been reported in the literature as OSCC or malignant oral GCT that resulted in unnecessary aggressive treatment for the affected patients. The aim of this study was to investigate if the cytokeratin pattern of the PCH can help in differentiating GCT from oral SCC. To distinguish between these two entities, we examined 12 patient specimens of oral GCT-PCH and oral SCC histologically and via immunohistochemistry (IHC) for CK13, CK17 and P75. The results suggest that the cytokeratin profile of PCH is similar to that of oral SCC. Therefore, consideration of IHC findings for epithelial markers alone may lead to erroneous diagnosis; thus, the presence of the granular tumor underneath the PCH and its immunopositivity for P75 or other neural definition markers can be essential to identify the underlying tumor and exclude oral SCC. Finally we recommend more studies on the molecular biology of PCH to understand how it can mimic oral SCC histologically without harboring its malignant phenotype clinically, which could have significant translational potential for understanding invasive oral SCC.