• Title/Summary/Keyword: quantitative profiles

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Identification of Cisplatin-Resistance Associated Genes through Proteomic Analysis of Human Ovarian Cancer Cells and a Cisplatin-resistant Subline

  • Zhou, Jing;Wei, Yue-Hua;Liao, Mei-Yan;Xiong, Yan;Li, Jie-Lan;Cai, Hong-Bing
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.12
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    • pp.6435-6439
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    • 2012
  • Chemoresistance to cancer therapy is a major obstacle to the effective treatment of human cancers with cisplatin (DDP), but the mechanisms of cisplatin-resistance are not clear. In this study, we established a cisplatin-resistant human ovarian cancer cell line (COC1/DDP) and identified differentially expressed proteins related to cisplatin resistance. The proteomic expression profiles in COC1 before and after DDP treatment were examined using 2-dimensional electrophoresis technology. Differentially expressed proteins were identified using matrix-assisted laser desorption/ ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and high performance liquid chromatography-electrospray tandem MS (NanoUPLC-ESI-MS/MS). 5 protein spots, for cytokeratin 9, keratin 1, deoxyuridine triphosphatase (dUTPase), aarF domain containing kinase 4 (ADCK 4) and cofilin1, were identified to be significantly changed in COC1/DDP compared with its parental cells. The expression of these five proteins was further validated by quantitative PCR and Western blotting, confirming the results of proteomic analysis. Further research on these proteins may help to identify novel resistant biomarkers or reveal the mechanism of cisplatin-resistance in human ovarian cancers.

Differential microRNA Expression by Solexa Sequencing in the Sera of Ovarian Cancer Patients

  • Ji, Ting;Zheng, Zhi-Guo;Wang, Feng-Mei;Xu, Li-Jian;Li, Lu-Feng;Cheng, Qi-Hui;Guo, Jiang-Feng;Ding, Xian-Feng
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.4
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    • pp.1739-1743
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    • 2014
  • MicroRNAs are a class of small noncoding RNA which play important regulatory roles in a variety of cancers. MiRNA-specific expression profiles have been reported for several pathological conditions. In this study, we combined large scale parallel Solexa sequencing to identify 11 up-regulated miRNAs and 19 down-regulated miRNAs with computational techniques in the sera of ovarian cancer patients while using healthy serum as the control. Among the above, four miRNAs (miR-22, miR-93, miR-106b, miR-451) were validated by quantitative RT-PCR and found to be significantly aberrantly expressed in the serum of ovarian cancer patients (P<0.05). There were no significant differences between samples from cancer stage I/II and III/IV. However, the levels of miR-106b (p=0.003) and miR-451 (p=0.007) were significantly different in those patients under and over 51 yearsof age. MiR-451 and miR-93 were also specific when analyzed with reference to different levels of CA125. This study shows that Solexa sequencing provides a promising method for cancer-related miRNA profiling, and selectively expressed miRNAs could be used as potential serum-based biomarkers for ovarian cancer diagnosis.

The mitochondrial proteome analysis in wheat roots

  • Kim, Da-Eun;Roy, Swapan Kumar;Kamal, Abu Hena Mostafa;Kwon, Soo Jeong;Cho, Kun;Cho, Seong-Woo;Park, Chul-Soo;Woo, Sun-Hee
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.126-126
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    • 2017
  • Mitochondria are important in wheat, as in all crops, as the main source of ATP for cell maintenance and growth including vitamin synthesis, amino acid metabolism and photorespiration. To investigate the mitochondrial proteome of the roots of wheat seedlings, a systematic and targeted analysis were carried out on the mitochondrial proteome from 15 day-old wheat seedling root material. Mitochondria were isolated by Percoll gradient centrifugation; and extracted proteins were separated and analyzed by Tricine SDS-PAGE along with LTQ-FTICR mass spectrometry. From the isolated the sample, 184 proteins were identified which is composed of 140 proteins as mitochondria and 44 proteins as other subcellular proteins that are predicted by the freeware subcellular predictor. The identified proteins in mitochondria were functionally classified into 12 classes using the ProtFun 2.2 server based on biological processes. Proteins were shown to be involved in amino acid biosynthesis (17.1%), biosynthesis of cofactors (6.4%), cell envelope (11.4%), central intermediary metabolism (10%), energy metabolism (20%), fatty acid metabolism (0.7%), purines and pyrimidines (5.7%), regulatory functions (0.7%), replication and transcription (1.4%), translation (22.1%), transport and binding (1.4%), and unknown (2.8%). These results indicate that many of the protein components present and functions of identifying proteins are common to other profiles of mitochondrial proteins performed to date. This dataset provides the first extensive picture, to our knowledge, of mitochondrial proteins from wheat roots. Future research is required on quantitative analysis of the wheat mitochondrial proteomes at the spatial and developmental level.

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Ultrastructural Analysis of Chemical Synapses in Cultured Wild Type Drosophila Embryonic Neurons (초파리 배자 신경세포의 화학적 신경연접 미세구조)

  • Oh, Hyun-Woo;Park, Ho-Yong
    • Applied Microscopy
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    • v.34 no.4
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    • pp.223-230
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    • 2004
  • To identify the structural basis of mutations that affect synaptic transmission we have begun quantitative ultrastructural descriptions of synapses in cultured Drosophila embryonic neurons. In wild-type cultures, synapses are distinguished by the parallel arrangement of a thickened pre- and post synaptic membrane separated by a synaptic cleft. The presynaptic active zones and postsynaptic densities are defined by electron dense material close to the membrane. Presynaptic regions are also characterized by the presence of one or more electron dense regions, presynaptic densities, around which a variable number of small, clear core synaptic vesicles (mean $35.1{\pm}1.44$ nm in diameter) are clustered. Subsets of these vesicles are in direct contact with either the presynaptic density or the membrane and are considered morphologically docked. A small number of larger, dense core vesicles are also observed in most presynaptic profiles.

Comparison of Trichothecene Biosynthetic Gene Expression between Fusarium graminearum and Fusarium asiaticum

  • Lee, Theresa;Lee, Seung-Ho;Shin, Jean Young;Kim, Hee-Kyoung;Yun, Sung-Hwan;Kim, Hwang-Yong;Lee, Soohyung;Ryu, Jae-Gee
    • The Plant Pathology Journal
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    • v.30 no.1
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    • pp.33-42
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    • 2014
  • Nivalenol (NIV) and deoxynivalenol (DON) are predominant Fusarium-producing mycotoxins found in grains, which are mainly produced by Fusarium asiaticum and F. graminearum. NIV is found in most of cereals grown in Korea, but the genetic basis for NIV production by F. asiaticum has not been extensively explored. In this study, 12 genes belonging to the trichothecene biosynthetic gene cluster were compared at the transcriptional level between two NIV-producing F. asiaticum and four DON-producing F. graminearum strains. Chemical analysis revealed that time-course toxin production patterns over 14 days did not differ between NIV and DON strains, excluding F. asiaticum R308, which was a low NIV producer. Both quantitative real-time polymerase chain reaction and Northern analysis revealed that the majority of TRI gene transcripts peaked at day 2 in both NIV and DON producers, which is 2 days earlier than trichothecene accumulation in liquid medium. Comparison of the gene expression profiles identified an NIV-specific pattern in two transcription factor-encoding TRI genes (TRI6 and TRI10) and TRI101, which showed two gene expression peaks during both the early and late incubation periods. In addition, the amount of trichothecenes produced by both DON and NIV producers were correlated with the expression levels of TRI genes, regardless of the trichothecene chemotypes. Therefore, the reduced production of NIV by R308 compared to NIV or DON by the other strains may be attributable to the significantly lower expression levels of the TRI genes, which showed early expression patterns.

Molecular Characterization and Expression Analysis of Adrenergic Receptor Beta 2 (ADRB2) Gene before and after Exercise in the Horse

  • Cho, Hyun-Woo;Shin, Sangsu;Song, Ki-Duk;Park, Jeong-Woong;Choi, Jae-Young;Lee, Hak-Kyo;Cho, Byung-Wook
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.5
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    • pp.686-690
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    • 2015
  • The adrenergic receptor beta 2 (ADRB2) plays a role in various physiological responses of the muscle to exercise, such as contraction and relaxation. Given its important role in muscle function, we investigated the structure of the horse ADRB2 gene and its expression pattern after exercise to determine if it can serve as a putative biomarker for recovery. Evolutionary analyses using synonymous and non-synonymous mutation ratios, were compared with other species (human, chimpanzee, mouse, rat, cow, pig, chicken, dog, and cat), and revealed the occurrence of positive selection in the horse ADRB2 gene. In addition, expression analyses by quantitative polymerase chain reaction exhibited ubiquitous distribution of horse ADRB2 in various tissues including lung, skeletal muscle, kidney, thyroid, appendix, colon, spinal cord and heart, with the highest expression observed in the lung. The expression of ADRB2 in skeletal muscle was significantly up-regulated about four folds 30 minutes post-exercise compared to pre-exercise. The expression level of ADRB2 in leukocytes, which could be collected with convenience compared with other tissues in horse, increased until 60 min after exercise but decreased afterward until 120 min, suggesting the ADRB2 expression levels in leukocytes could be a useful biomarker to check the early recovery status of horse after exercise. In conclusion, we identified horse ADRB2 gene and analyzed expression profiles in various tissues. Additionally, analysis of ADBR2 gene expression in leukocytes could be a useful biomarker useful for evaluation of early recovery status after exercise in racing horses.

Computerized Tomographic Measurements of Morphometric Parameters of the C2 for the Feasibility of Laminar Screw Fixation in Korean Population

  • Kim, Young-June;Rhee, Woo-Tack;Lee, Sang-Bok;You, Seung-Hoon;Lee, Sang-Youl
    • Journal of Korean Neurosurgical Society
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    • v.44 no.1
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    • pp.15-18
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    • 2008
  • Objective : C2 laminar screw fixation is considered as an excellent alternative to Magerl's transfacetal approach or Harms construct for the atlantoaxial stabilization. However, to our knowledge, there is no report on the feasibility of the new approach to Korean population. We investigated morphometric parameters of the dorsal arch of the C2 to provide the quantitative data for the feasibility of laminar screw fixation. Methods : One-hundred-and-two patients' cervical computed tomography had been reconstructed and investigated on the anatomical parameters related with C2 laminar screw placement. Sixty patients were male and forty-two patients were female. Measurements included the laminar thickness and slope, spino-laminar angle, and maximal screw length. Results : Ages ranged from 20 to 81 and the mean age was 48.4. Mean laminar thickness was 5.7 mm (${\pm}1.0$) (5.8 mm in male and 5.4 mm in female). Fifty-one patients (50%) had a laminar thickness smaller than 5.5 mm at least unilaterally, therefore the patients were considered as inappropriate candidates for the laminar screw fixation in the smaller side of the laminae. Mean value of maximal length of screw was 33.3 mm (34.3 mm in male and 31.9 mm in female). Mean spino-laminar angle was $43.2^{\circ}$ and mean slope angle was $32.9^{\circ}$. Conclusion : Half of patients had inappropriate laminar profiles to accommodate a 3.5 mm screw in at least one side of the axis. The three-dimensional computed tomography reconstruction is mandatory for the preoperative assessment for the feasibility of the C2 lamina.

The Dose Dependent Effects of Ruxolitinib on the Invasion and Tumorigenesis in Gliomas Cells via Inhibition of Interferon Gamma-Depended JAK/STAT Signaling Pathway

  • Delen, Emre;Doganlar, Oguzhan
    • Journal of Korean Neurosurgical Society
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    • v.63 no.4
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    • pp.444-454
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    • 2020
  • Objective : Glioblastoma multiforme (GBM) is the most aggressive for of brain tumor and treatment often fails due to the invasion of tumor cells into neighboring healthy brain tissues. Activation of the Janus kinase-signal transducer and activator of transcription (JAK/STAT) signaling pathway is essential for normal cellular function including angiogenesis, and has been proposed to have a pivotal role in glioma invasion. This study aimed to determine the dose-dependent effects of ruxolitinib, an inhibitor of JAK, on the interferon (IFN)-I/IFN-α/IFN-β receptor/STAT and IFN-γ/IFN-γ receptor/STAT1 axes of the IFN-receptor-dependent JAK/STAT signaling pathway in glioblastoma invasion and tumorigenesis in U87 glioblastoma tumor spheroids. Methods : We administered three different doses of ruxolitinib (50, 100, and 200 nM) to human U87 glioblastoma spheroids and analyzed the gene expression profiles of IFNs receptors from the JAK/STAT pathway. To evaluate activation of this pathway, we quantified the phosphorylation of JAK and STAT proteins using Western blotting. Results : Quantitative real-time polymerase chain reaction analysis demonstrated that ruxolitinib led to upregulated of the IFN-α and IFN-γ while no change on the hypoxia-inducible factor-1α and vascular endothelial growth factor expression levels. Additionally, we showed that ruxolitinib inhibited phosphorylation of JAK/STAT proteins. The inhibition of IFNs dependent JAK/STAT signaling by ruxolitinib leads to decreases of the U87 cells invasiveness and tumorigenesis. We demonstrate that ruxolitinib may inhibit glioma invasion and tumorigenesis through inhibition of the IFN-induced JAK/STAT signaling pathway. Conclusion : Collectively, our results revealed that ruxolitinib may have therapeutic potential in glioblastomas, possibly by JAK/STAT signaling triggered by IFN-α and IFN-γ.

Improvement of ${\beta}-glucosidase$ Activity of Olea europaea Fruit Extracts Processed by Membrane Technology

  • Mazzei, R.;Giomo, L.;Spadafora, A.;Mazzuca, S.;Drioli, E.
    • Korean Membrane Journal
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    • v.8 no.1
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    • pp.58-66
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    • 2006
  • The ${\beta}-glucosidase$ from olive fruit is of particular interest compared to the ones from other sources because it has shown to have high specifity to convert the oleuropein into dialdehydes, which have antibacterial activity and are of high interest for their application in the food and pharmaceutical fields. The enzyme is not yet commercially available and advanced clean and safe technologies for its purification able to maintain the functional stability are foreseen. The purification of this protein from fruit extracts has been already tempted by electrophoresis but either enzyme deactivation or high background with unclear profiles occurred. In this work, fruit extracts obtained from the ripening stage that showed the highest enzyme activity have been processed by diafiltration and ultrafiltration. Asymmetric membranes made of polyamide or polysulphone having 50 and 30 kDa molecular weight cut-off, respectively, were tested for the diafiltration process. Ultrafiltration membranes made of polyethersulfone with 4 kDa molecular weight cut-off were used to concentrate the dia-filtered permeate solutions. The efficiency of the separation processes was evaluated byenzyme activity tests using the hydrolysis of p-D-nitrophenyl-${\beta}$-D-glucopyranoside (pNPGlc) as reaction model. Qualitative and quantitative electrophoresis were applied to analyze the composition of protein solution before and after the membrane separation; in addition dot blot and western blot analyses were applied to verify the presence of ${\beta}-glucosidase$ in the processed fractions. The overall results showed that the ${\beta}-glucosidase$ functional stability was preserved during the membrane operations and the removal of 20 kDa proteins allowed to increase the specific activity of the enzyme of about 52% compared to the one present in the initial fruit extract.

Structure and Property Analysis of Nanoporous Low Dielectric Constant SiCOH Thin Films

  • Heo, Gyu-Yong;Lee, Mun-Ho;Lee, Si-U;Park, Yeong-Hui
    • Proceedings of the Korean Institute of Surface Engineering Conference
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    • 2009.05a
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    • pp.167-169
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    • 2009
  • We have carried out quantitative structure and property analysis of the nanoporous structures of low dielectric constant (low-k) carbon-doped silicon oxide (SiCOH) films, which were deposited with plasma enhanced chemical vapor deposition (PECVD) using vinyltrimethylsilane (VTMS), divinyldimethylsilane (DVDMS), and tetravinylsilane (TVS) as precursor and oxygen as an oxidant gas. We found that the SiCOH film using VTMS only showed well defined spherical nanopores within the film after thermal annealing at $450^{\circ}C$ for 4 h. The average pore radius of the generated nanopores within VTMS SiCOH film was 1.21 nm with narrow size distribution of 0.2. It was noted that thermally labile $C_{x}H_{y}$ phase and Si-$CH_3$ was removed to make nanopore within the film by thermal annealing. Consequently, this induced that decrease of average electron density from 387 to $321\;nm^{-3}$ with increasing annealing temperature up to $450^{\circ}C$ and taking a longer annealing time up to 4 h. However, the other SiCOH films showed featureless scattering profiles irrespective of annealing conditions and the decreases of electron density were smaller than VTMS SiCOH film. Because, with more vinyl groups are introduced in original precursor molecule, films contain more organic phase with less volatile characteristic due to the crosslinking of vinyl groups. Collectively, the presenting findings show that the organosilane containing vinyl group was quite effective to deposit SiCOH/$C_{x}H_{y}$ dual phase films, and post annealing has an important role on generation of pores with the SiCOH film.

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