• Title/Summary/Keyword: Differential expression analysis

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Differential gene expression analysis of human cumulus cells

  • Demiray, Sirin Bakti;Goker, Ege Nazan Tavmergen;Tavmergen, Erol;Yilmaz, Ozlem;Calimlioglu, Nilufer;Soykam, Huseyin Okan;Oktem, Gulperi;Sezerman, Ugur
    • Clinical and Experimental Reproductive Medicine
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    • v.46 no.2
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    • pp.76-86
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    • 2019
  • Objective: This study was performed to explore the possibility that each oocyte and its surrounding cumulus cells might have different genetic expression patterns that could affect human reproduction. Methods: Differential gene expression analysis was performed for 10 clusters of cumulus cells obtained from 10 cumulus-oocyte complexes from 10 patients. Same procedures related to oocyte maturation, microinjection, and microarray analyses were performed for each group of cumulus cells. Two differential gene expression analyses were performed: one for the outcome of clinical pregnancy and one for the outcome of live birth. Results: Significant genes resulting from these analyses were selected and the top 20 affected pathways in each group were analyzed. Circadian entrainment is determined to be the most affected pathway for clinical pregnancy, and proteoglycans in cancer pathway is the most affected pathway for live birth. Circadian entrainment is also amongst the 12 pathways that are found to be in top 20 affected pathways for both outcomes, and has both lowest p-value and highest number of times found count. Conclusion: Although further confirmatory studies are necessary, findings of this study suggest that these pathways, especially circadian entrainment in cumulus cells, may be essential for embryo development and pregnancy.

Gene Microarray Assessment of Multiple Genes and Signal Pathways Involved in Androgen-dependent Prostate Cancer Becoming Androgen Independent

  • Liu, Jun-Bao;Dai, Chun-Mei;Su, Xiao-Yun;Cao, Lu;Qin, Rui;Kong, Qing-Bo
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.22
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    • pp.9791-9795
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    • 2014
  • To study the gene expression change and possible signal pathway during androgen-dependent prostate cancer (ADPC) becoming androgen-independent prostate cancer (AIPC), an LNCaP cell model of AIPC was established using flutamide in combination with androgen-free environment inducement, and differential expression genes were screened by microarray. Then the biological process, molecular function and KEGG pathway of differential expression genes are analyzed by Molecule Annotation System (MAS). By comparison of 12,207 expression genes, 347 expression genes were acquired, of which 156 were up-ragulated and 191 down-regulated. After analyzing the biological process and molecule function of differential expression genes, these genes are found to play crucial roles in cell proliferation, differntiation, cell cycle control, protein metabolism and modification and other biological process, serve as signal molecules, enzymes, peptide hormones, cytokines, cytoskeletal proteins and adhesion molecules. The analysis of KEGG show that the relevant genes of AIPC transformation participate in glutathione metabolism, cell cycle, P53 signal pathway, cytochrome P450 metabolism, Hedgehog signal pathway, MAPK signal pathway, adipocytokines signal pathway, PPAR signal pathway, TGF-${\beta}$ signal pathway and JAK-STAT signal pathway. In conclusion, during the process of ADPC becoming AIPC, it is not only one specific gene or pathway, but multiple genes and pathways that change. The findings above lay the foundation for study of AIPC mechanism and development of AIPC targeting drugs.

Differential Expression of Rice Lipid Transfer Protein Gene (LTP) Classes in Response to γ-irradiation Pattern (감마선 조사 패턴에 따른 벼의 Lipid Transfer Protein Gene (LTP)의 발현 차이)

  • Kim, Sun-Hee;Song, Mira;Jang, Duk-Soo;Kang, Si-Yong;Kim, Jin-Baek;Kim, Sang Hoon;Ha, Bo-Keun;Park, Yong Dae;Kim, Dong Sub
    • Journal of Radiation Industry
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    • v.5 no.1
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    • pp.47-54
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    • 2011
  • In this study, we investigated to evaluate differential expression of genes encoding lipid transfer proteins (LTP) by acute and chronic gamma irradiation in rice. After acute and chronic gamma irradiation by 100 Gy and 400 Gy to rice plant, necrotic lesion was observed in the leaf blade and anthocyanin contents were increased. We isolated a total of 21 rice lipid transfer protein (LTP) genes in the TIGR database, and these genes were divided into four different groups on the basis of nucleotide sequences. The LTP genes also were classified as different four classes according to expression pattern using RT-PCR. Group A, B contained genes with increased expression and decreased expression in acute and chronic, respectively. Group C contained genes with contrasted expression pattern. Group D wasn't a regular pattern. But the specific affinity was not obtained between two grouping.

Analysis of Structure and Expression of Grapevine 2-oxoglutarate Oxygenase Genes in Response to Low Temperature

  • Kim, Seon Ae;Ahn, Soon Young;Yun, Hae Keun
    • Horticultural Science & Technology
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    • v.34 no.1
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    • pp.46-54
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    • 2016
  • 2-Oxoglutarate (2OG) acts as a signaling molecule and plays a critical role in secondary metabolism in a variety of organisms, including plants. Six 2-oxoglutarate (2OG) and Fe(II) oxygenase (2OGO) genes, VlCE2OGO1 [Vitis labruscana 2-oxoglutarate (2OG) and Fe(II) oxygenase 1], VlCE2OGO2, VlCE2OGO3, VlCE2OGO4, VlCE2OGO5, and VlCE2OGO6, which show different expression patterns upon transcriptome analysis of 'Campbell Early' grapevine exposed to low temperature for 4 weeks, were analyzed for their structure and expression. Comparison of the deduced amino acid sequences of the 2OGO genes from the V. labruscana transcripts revealed sequence similarities of 38.6% (VlCE2OGO1 and VlCE2OGO2) to 19.2% (VlCE2OGO2 and VlCE2OGO3). The lengths of these genes ranged from 1053 to 2298 bp, and they encoded 316 to 380 amino acids. The prediction of the secondary structure of the encoded proteins by Self-Optimized Prediction Method with Alignment (SOPMA) indicated that all the genes contained alpha helix (23.95 to 41.71%), extended strand (16 to 22.34%), beta turn (6.65 to 9.22%), and random coil (32.97 to 51.58%) in the analysis. Specific primers from unique regions in each gene obtained by alignment of nucleotide sequences were used in real time PCR for analysis of gene expression. All tested genes showed differential expression in grapevines exposed to low temperature. Of the six transcripts, VlCE2OGO1, VlCE2OGO2, and VlCE2OGO3 were up-regulated and VlCE2OGO4, VlCE2OGO5, and VlCE2OGO6 were down-regulated in response to cold treatments at all tested time points. The 2OG genes can be used for elucidation of mechanisms of tolerance to cold and as valuable molecular genetic resources for selection in breeding programs for cold-hardy grapevines.

Comparative Analysis of Gene Expression in the Female Reproductive Organs

  • Kim, Min-Goo;Seo, Hee-Won;Choi, Yo-Han;Lee, Chang-Kyu;Ka, Hak-Hyun
    • Journal of Embryo Transfer
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    • v.24 no.2
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    • pp.77-87
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    • 2009
  • To understand molecular and cellular mechanisms of many gene products in the female reproductive organs including the ovary and uterine endometrium as well as during embryo development, researchers have developed and utilized many effective methodologies to analyze gene expression in cells, tissues and animals over the last several decades. For example, blotting techniques have helped to understand molecular functions at DNA, RNA and protein levels, and the reverse transcription-polymerase chain reaction (RT-PCR) method has been widely used in gene expression analysis. However, some conventional methods are not sufficient to understand regulation and function of genes expressed in very complex patterns in many organs. Thus, it is required to adopt more high-throughput and reliable techniques. Here, we describe several techniques used widely recently to analyze gene expression, including annealing control based-PCR, differential display-PCR, expressed sequence tag, suppression subtractive hybridization and microarray techniques. Use of these techniques will help to analyze expression pattern of many genes from small scale to large scale and to compare expression patterns of genes in one sample to another. In this review, we described principles of these methodologies and summarized examples of comparative analysis of gene expression in female reproductive organs with help of those methodologies.

Characterization of tissue-specific mbu-3 gene expression in the mouse central nervous system

  • Lee, Chae-Jin;Cho, Eun-Young;Kim, Sun-Jung
    • BMB Reports
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    • v.41 no.12
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    • pp.875-880
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    • 2008
  • Mbu-3 is a novel mouse brain unigene that was identified by digital differential display. In this study, expression of the gene was chased through developmental stages and the protein product was identified in the brain. The cDNA sequence was 3,995-bp long and contained an ORF of 745 AA. Database searches revealed that the chicken SST273 gene containing LRR- and Ig-domain was an mbu-3 orthologue. Tissue specificity for the gene was examined in embryos and in brains at post-natal and adult stages. During the embryonic stages, mbu-3 was localized to the central nervous system in the brain and spinal cord. In the early post-natal stages, the gene was evenly expressed in the brain. However, with aging, expression was confined to specific regions, particularly the hippocampus. The protein was approximately 95 kDa as determined by Western blot analysis of brain extracts.

Differentially Expressed Genes in Hemocytes of Vibrio harveyi-challenged Shrimp Penaeus monodon

  • Somboonwiwat, Kunlaya;Supungul, Premruethai;Rimphanitchayakit, Vichien;Aoki, Takashi;Hirono, Ikuo;Tassanakajon, Anchalee
    • BMB Reports
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    • v.39 no.1
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    • pp.26-36
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    • 2006
  • Differential Display PCR technique (DD-PCR) was used for the analysis of altered gene expression in hemocytes of Vibrio harveyi-infected Penaeus monodon. Forty-four combinations of arbitrary and oligo(dT) primers were used to screen for differentially expressed genes. A total of 79 differentially expressed bands could be identified from 33 primer combinations. These included 48 bands (61%) whose expression level increased and 31 bands (39%) decreased after V. harveyi challenge. Subsequently, forty-eight differential display fragments were successfully reamplified and cloned. A total of 267 clones were randomly selected and sequenced. The sequence analysis showed that 85 (31%) out of 267 clones were matched with sequences in the GenBank database which represented 24 different genes with known functions. Among the known genes, glucose transporter 1, interferon-related developmental regulator 1, lysozyme, profilin, SERPINB3, were selected for further confirmation of their differentially expression patterns by real-time PCR. The results showed increasing in expression level of the selected genes in shrimp hemocytes after microbial challenge suggesting the involvement of such genes in bacterial response in shrimp. The anti-lipopolysaccharide factor type 3 (ALFPm3) gene, previously reported in P. monodon (Supungul et al., 2002) was found among the up-regulated genes but diversity due to amino acid changes was observed. Increase in ALFPm3 transcripts upon V. harveyi injection is in accordance with that found in the previous study.

Differential Expression of Multiple Connexins in Rat Corpus and Cauda Epididymis at Various Postnatal Stages

  • Lee, Ki-Ho
    • Journal of Animal Science and Technology
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    • v.55 no.6
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    • pp.521-530
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    • 2013
  • Direct cell-cell communication via the transfer of small molecules between neighboring cells in tissue is accomplished by gap junctions composed of various connexins (Cxs). Proper postnatal development of the epididymis is important for acquisition of male reproduction. The epididymal epithelium is composed of several cell types, and some of these cells are connected by gap junctions. The present study was conducted to determine the presence of Cx transcripts in the corpus and cauda epididymis. In addition, transcriptional changes of Cxs expressed during different postnatal stages were examined by real-time PCR analysis. In both epididymal regions, the same nine Cx transcripts of thirteen Cxs tested were detected. In the corpus epididymis, the highest levels of Cxs31.1 and 37 transcripts were observed at 45 days of age, and amounts of Cxs26, 30.3, and 32 transcripts increased with age and subsequently decreased in the elderly. Expression of Cx31 was greatly increased in the adult and elder stages, while Cxs40, 43, and 45 were abundant in the early postnatal stages. In the cauda epididymis, expression of Cxs26, 30.3, 31.1, 37, and 40 reached the highest levels at 5 months of age. The levels of Cxs31 and 32 mRNAs fluctuated throughout the postnatal period. The amounts of Cxs43 and 45 transcripts were more abundant during the late neonatal and prepubertal ages than later ages. These findings suggest that regional specification of the epididymis is partly regulated by differential expression of Cx genes during the postnatal developmental period.

Analysis of Different Activation Statuses of Human Mammary Epithelial Cells from Young and Old Groups

  • Feng, Chen-Chen;Chen, Li-Na;Chen, Mei-Jun;Li, Wan;Jia, Xu;Zhou, Yan-Yan;He, Wei-Ming
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.8
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    • pp.3763-3766
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    • 2014
  • Human mammary epithelial cells have different proliferative statuses and demonstrate a close relationship with age and cell proliferation. Research on this topic could help understand the occurrence, progression and prognosis of breast cancer. In this article, using significance analysis of a microarray algorithm, we analyzed gene expression profiles of human mammary epithelial cells of different proliferative statuses and different age groups. The results showed there were significant differences in gene expression in the same proliferation status between elderly and young groups. Three common differentially expressed genes were found to dynamically change with the proliferation status and to be closely related to tumorigenesis. We also found elderly group had less status-related differential genes from actively proliferating status to intermediate status and more statusrelated differential genes from intermediate status than the young group. Finally, functional enrichment analyses allowed evaluation of the detailed roles of these differentially-expressed genes in tumor progression.