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DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 俞松良 | |
dc.contributor.author | Yu-Wen Tseng | en |
dc.contributor.author | 曾郁雯 | zh_TW |
dc.date.accessioned | 2021-06-16T06:52:09Z | - |
dc.date.available | 2022-07-31 | |
dc.date.copyright | 2014-10-09 | |
dc.date.issued | 2014 | |
dc.date.submitted | 2014-07-22 | |
dc.identifier.citation | 1. Fabrega A, Vila J: Salmonella enterica serovar Typhimurium skills to succeed in the host: virulence and regulation. Clinical microbiology reviews 2013, 26(2):308-341.
2. Gordon MA: Invasive nontyphoidal Salmonella disease: epidemiology, pathogenesis and diagnosis. Current opinion in infectious diseases 2011, 24(5):484-489. 3. Haraga A, Ohlson MB, Miller SI: Salmonellae interplay with host cells. Nature reviews Microbiology 2008, 6(1):53-66. 4. Foster JW, Hall HK: Inducible pH homeostasis and the acid tolerance response of Salmonella typhimurium. Journal of Bacteriology 1991, 173:5129-5135. 5. McGhie EJ, Brawn LC, Hume PJ, Humphreys D, Koronakis V: Salmonella takes control: effector-driven manipulation of the host. Current opinion in microbiology 2009, 12(1):117-124. 6. Francis. CL, Starnbach. MN, Falkow. S: Morphological and cytoskeletal changes in epithelial cells occur immediately upon interaction with Salmonella typhimurium grown under low-oxygen conditions. Molecular Microbiology 1992, 6:3077-3087. 7. Hansen-Wester I, Hensel M: Salmonella pathogenicity islands encoding type III secretion systems. Microbes and Infection 2001, 3(7):549-559. 8. Hicks SW, Galan JE: Exploitation of eukaryotic subcellular targeting mechanisms by bacterial effectors. Nat Rev Micro 2013, 11(5):316-326. 9. Peter J. Christie, Krishnamohan Atmakuri, Vidhya Krishnamoorthy, Simon Jakubowski, Cascales E: Biogenesis, Architecture, and Function of Bacterial Type IV Secretion Systems. The Annual Review of Microbiology 2005:451-485. 10. Galan JE, Wolf-Watz H: Protein delivery into eukaryotic cells by type III secretion machines. Nature 2006, 444(7119):567-573. 11. Marsman M, Jordens I, Kuijl C, Janssen L, Neefjes J: Dynein-mediated vesicle transport controls intracellular Salmonella replication. Molecular biology of the cell 2004, 15(6):2954-2964. 12. de Jong HK, Parry CM, van der Poll T, Wiersinga WJ: Host–Pathogen Interaction in Invasive Salmonellosis. PLoS Pathog 2012, 8(10):e1002933. 13. Galan JE: Interaction of Salmonella with host cells through the centisome 63 type III secretion system. Current opinion in microbiology 1999, 2(1):46-50. 14. Tierrez A, Garcia-del Portillo F: New concepts in Salmonella virulence: the importance of reducing the intracellular growth rate in the host. Cellular microbiology 2005, 7(7):901-909. 15. Haneda T, Ishii Y, Shimizu H, Ohshima K, Iida N, Danbara H, Okada N: Salmonella type III effector SpvC, a phosphothreonine lyase, contributes to reduction in inflammatory response during intestinal phase of infection. Cellular microbiology 2012, 14(4):485-499. 16. n CRB, Salcedo SP, Holden DW: Growth and killing of a Salmonella enterica serovar Typhimurium sifA mutant strain in the cytosol of different host cell lines. Microbiology 2002, 148:2705–2715. 17. Figureueira R, Holden DW: Functions of the Salmonella pathogenicity island 2 (SPI-2) type III secretion system effectors. Microbiology 2012, 158(Pt 5):1147-1161. 18. Poh J, Odendall C, Spanos A, Boyle C, Liu M, Freemont P, Holden DW: SteC is a Salmonella kinase required for SPI-2-dependent F-actin remodelling. Cellular microbiology 2008, 10(1):20-30. 19. Gorvel J-P, Meresse S: Maturation steps of the Salmonella-containing vacuole. Microbes and Infection 2001, 3(14–15):1299-1303. 20. Lamkanfi M, Dixit VM: Manipulation of host cell death pathways during microbial infections. Cell host & microbe 2010, 8(1):44-54. 21. Kim M, Ashida H, Ogawa M, Yoshikawa Y, Mimuro H, Sasakawa C: Bacterial interactions with the host epithelium. Cell host & microbe 2010, 8(1):20-35. 22. Ashida H, Ogawa M, Kim M, Mimuro H, Sasakawa C: Bacteria and host interactions in the gut epithelial barrier. Nature chemical biology 2012, 8(1):36-45. 23. Loktionov A: Cell exfoliation in the human colon: Myth, reality and implications for colorectal cancer screening. International Journal of Cancer 2007, 120(11):2281-2289. 24. Amcheslavsky A, Jiang J, Ip YT: Tissue Damage-Induced Intestinal Stem Cell Division in Drosophila. Cell stem cell 2009, 4(1):49-61. 25. Jiang H, Patel PH, Kohlmaier A, Grenley MO, McEwen DG, Edgar BA: Cytokine/Jak/Stat Signaling Mediates Regeneration and Homeostasis in the Drosophila Midgut. Cell 2009, 137(7):1343-1355. 26. Iwai H, Kim M, Yoshikawa Y, Ashida H, Ogawa M, Fujita Y, Muller D, Kirikae T, Jackson PK, Kotani S et al: A Bacterial Effector Targets Mad2L2, an APC Inhibitor, to Modulate Host Cell Cycling. Cell 2007, 130(4):611-623. 27. Mimuro H, Suzuki T, Nagai S, Rieder G, Suzuki M, Nagai T, Fujita Y, Nagamatsu K, Ishijima N, Koyasu S et al: Helicobacter pylori Dampens Gut Epithelial Self-Renewal by Inhibiting Apoptosis, a Bacterial Strategy to Enhance Colonization of the Stomach. Cell host & microbe 2007, 2(4):250-263. 28. Valdez Y, Ferreira RR, Finlay BB: Molecular Mechanisms of Salmonella Virulence and Host Resistance. In Molecular Mechanisms of Bacterial Infection via the Gut. Edited by Sasakawa C, vol. 337: Springer Berlin Heidelberg; 2009: 93-127. Current Topics in Microbiology and Immunology. 29. Albiger B, Dahlberg S, Henriques-Normark B, Normark S: Role of the innate immune system in host defence against bacterial infections: focus on the Toll-like receptors. Journal of internal medicine 2007, 261(6):511-528. 30. J Philpott D, E Girardin S, J Sansonetti P: Innate immune responses of epithelial cells following infection with bacterial pathogens. Current Opinion in Immunology 2001, 13(4):410-416. 31. Puja Vora, Adrienne Youdim, Lisa S. Thomas, Masayuki Fukata, Samuel Y. Tesfay, Katie Lukasek, Kathrin S. Michelsen, Akihiro Wada, Toshiya Hirayama, Moshe Arditi et al: Beta-Defensin-2 Expression Is Regulated by TLR Signaling in Intestinal Epithelial Cells. The Journal of Immunology 2004, 173:5398-5405. 32. Tosi MF: Innate immune responses to infection. Journal of Allergy and Clinical Immunology 2005, 116(2):241-249. 33. Mostowy S: Autophagy and bacterial clearance: a not so clear picture. Cellular microbiology 2013, 15(3):395-402. 34. Choi AM, Ryter SW, Levine B: Autophagy in human health and disease. The New England journal of medicine 2013, 368(7):651-662. 35. Dorn BR, Dunn WA, Progulske-Fox A: Bacterial interactions with the autophagic pathway. Cellular microbiology 2002, 4(1):1-10. 36. Rubinsztein DC, Codogno P, Levine B: Autophagy modulation as a potential therapeutic target for diverse diseases. Nature reviews Drug discovery 2012, 11(9):709-730. 37. Levine B, Mizushima N, Virgin HW: Autophagy in immunity and inflammation. Nature 2011, 469(7330):323-335. 38. Rich KA, Burkett C, Webster P: Cytoplasmic bacteria can be targets for autophagy. Cellular microbiology 2003, 5(7):455-468. 39. Birmingham CL, Higgins DE, Brumell JH: Avoiding death by autophagy: Interactions of Listeria monocytogenes with the macrophage autophagy system. Autophagy 2008, 4(3):368-371. 40. Haglund CM, Welch MD: Pathogens and polymers: microbe-host interactions illuminate the cytoskeleton. The Journal of cell biology 2011, 195(1):7-17. 41. Fabri M, Stenger S, Shin DM, Yuk JM, Liu PT, Realegeno S, Lee HM, Krutzik SR, Schenk M, Sieling PA et al: Vitamin D is required for IFN-gamma-mediated antimicrobial activity of human macrophages. Science translational medicine 2011, 3(104):104ra102. 42. Birmingham CL, Smith AC, Bakowski MA, Yoshimori T, Brumell JH: Autophagy controls Salmonella infection in response to damage to the Salmonella-containing vacuole. The Journal of biological chemistry 2006, 281(16):11374-11383. 43. Benjamin Jamaal L, Sumpter Jr R, Levine B, Hooper Lora V: Intestinal Epithelial Autophagy Is Essential for Host Defense against Invasive Bacteria. Cell host & microbe 2013, 13(6):723-734. 44. Frankel LB, Lund AH: MicroRNA regulation of autophagy. Carcinogenesis 2012, 33(11):2018-2025. 45. Fu L-l, Wen X, Bao J-k, Liu B: MicroRNA-modulated autophagic signaling networks in cancer. The International Journal of Biochemistry & Cell Biology 2012, 44(5):733-736. 46. Hua Zhu, Hao Wu, Xiuping Liu, Biao Li, Yun Chen, Xingcong Ren, Chang-Gong Liu, Yang J-M: Regulation of autophagy by a beclin 1-targeted MicroRNA, miR-30a, in cancer cells. Autophagy 2009, 5:816-823. 47. Montagner S, Orlandi EM, Merante S, Monticelli S: The role of miRNAs in mast cells and other innate immune cells. Immunological Reviews 2013, 253(1):12-24. 48. Bartel DP: MicroRNAs: Genomics, Biogenesis, Mechanism, and Function. Cell 2004, 116(2):281-297. 49. Guo L, Lu Z: The Fate of miRNA* Strand through Evolutionary Analysis: Implication for Degradation As Merely Carrier Strand or Potential Regulatory Molecule? PLoS ONE 2010, 5(6):e11387. 50. Bronevetsky Y, Ansel KM: Regulation of miRNA biogenesis and turnover in the immune system. Immunological Reviews 2013, 253(1):304-316. 51. Alvarez-Garcia I, Miska EA: MicroRNA functions in animal development and human disease. Development 2005, 132(21):4653-4662. 52. Delić D, Dkhil M, Al-Quraishy S, Wunderlich F: Hepatic miRNA expression reprogrammed by Plasmodium chabaudi malaria. Parasitol Res 2011, 108(5):1111-1121. 53. Zhou R, Hu G, Liu J, Gong A-Y, Drescher KM, Chen X-M: NF-kappaB p65-Dependent Transactivation of miRNA Genes following <italic>Cryptosporidium parvum</italic> Infection Stimulates Epithelial Cell Immune Responses. PLoS Pathog 2009, 5(12):e1000681. 54. Lecellier CH, Dunoyer P, Arar K, Lehmann-Che J, Eyquem S, Himber C, Saib A, Voinnet O: A cellular microRNA mediates antiviral defense in human cells. Science 2005, 308(5721):557-560. 55. Nathans R, Chu C-y, Serquina AK, Lu C-C, Cao H, Rana TM: Cellular MicroRNA and P Bodies Modulate Host-HIV-1 Interactions. Molecular cell 2009, 34(6):696-709. 56. Navarro L, Dunoyer P, Jay F, Arnold B, Dharmasiri N, Estelle M, Voinnet O, Jones JD: A plant miRNA contributes to antibacterial resistance by repressing auxin signaling. Science 2006, 312(5772):436-439. 57. Eulalio A, Schulte L, Vogel J: The mammalian microRNA response to bacterial infections. RNA Biology 2012, 9(6):742-750. 58. Oertli M, Engler DB, Kohler E, Koch M, Meyer TF, Muller A: MicroRNA-155 is essential for the T cell-mediated control of Helicobacter pylori infection and for the induction of chronic Gastritis and Colitis. Journal of immunology 2011, 187(7):3578-3586. 59. Voinnet O: Micro-balancing innate immunity to Salmonella. The EMBO journal 2011, 30(10):1877-1879. 60. Rusca N, Monticelli S: MiR-146a in Immunity and Disease. Molecular biology international 2011, 2011:437301. 61. Lu L-F, Boldin MP, Chaudhry A, Lin L-L, Taganov KD, Hanada T, Yoshimura A, Baltimore D, Rudensky AY: Function of miR-146a in Controlling Treg Cell-Mediated Regulation of Th1 Responses. Cell 2010, 142(6):914-929. 62. Li L, Chen XP, Li YJ: MicroRNA-146a and Human Disease. Scandinavian Journal of Immunology 2010, 71(4):227-231. 63. Nahid MA, Pauley KM, Satoh M, Chan EK: miR-146a is critical for endotoxin-induced tolerance: IMPLICATION IN INNATE IMMUNITY. The Journal of biological chemistry 2009, 284(50):34590-34599. 64. Taganov KD, Boldin MP, Chang KJ, Baltimore D: NF-kappaB-dependent induction of microRNA miR-146, an inhibitor targeted to signaling proteins of innate immune responses. Proceedings of the National Academy of Sciences of the United States of America 2006, 103(33):12481-12486. 65. Bhaumik D, Scott GK, Schokrpur S, Patil CK, Campisi J, Benz CC: Expression of microRNA-146 suppresses NF-kappaB activity with reduction of metastatic potential in breast cancer cells. Oncogene 2008, 27(42):5643-5647. 66. Rujuan Dai, Rebecca A. Phillips, Yan Zhang DK, Oswald Crasta, Ahmed SA: Suppression of LPS-induced Interferon-g and nitric oxide in splenic lymphocytes by select estrogen-regulated microRNAs: a novel mechanism of immune modulation. Blood 2008, 112:4591-4597. 67. Liu Z, Xiao B, Tang B, Li B, Li N, Zhu E, Guo G, Gu J, Zhuang Y, Liu X et al: Up-regulated microRNA-146a negatively modulate Helicobacter pylori-induced inflammatory response in human gastric epithelial cells. Microbes and Infection 2010, 12(11):854-863. 68. Schnitger AKD, Machova A, Mueller RU, Androulidaki A, Schermer B, Pasparakis M, Kronke M, Papadopoulou N: Listeria monocytogenes Infection in Macrophages Induces Vacuolar-Dependent Host miRNA Response. PLoS ONE 2011, 6(11):e27435. 69. Sharbati J, Lewin A, Kutz-Lohroff B, Kamal E, Einspanier R, Sharbati S: Integrated MicroRNA-mRNA-Analysis of Human Monocyte Derived Macrophages upon <italic>Mycobacterium avium</italic> subsp. <italic>hominissuis</italic> Infection. PLoS ONE 2011, 6(5):e20258. 70. Sharbati S, Sharbati J, Hoeke L, Bohmer M, Einspanier R: Quantification and accurate normalisation of small RNAs through new custom RT-qPCR arrays demonstrates Salmonella-induced microRNAs in human monocytes. BMC Genomics 2012, 13(1):1-11. 71. Giannella RA, Washington O, Gemski P, Formal SB: Invasion of HeLa Cells by Salmonella typhimurium: A Model for Study of Invasiveness of Salmonella. The Journal of Infectious Diseases 1973, 128(1):69-75. 72. Abraham VC, Taylor DL, Haskins JR: High content screening applied to large-scale cell biology. Trends in Biotechnology 2004, 22(1):15-22. 73. Zanella F, Lorens JB, Link W: High content screening: seeing is believing. Trends in Biotechnology 2010, 28(5):237-245. 74. Hariharan P: Apochromatic lens combinations: a novel design approach. Optics & Laser Technology 1997, 29(4):217-219. 75. Malik Kale P, Jolly CE, Lathrop S, Winfree S, Luterbach C, Steele-Mortimer O: Salmonella - at home in the host cell. Frontiers in Microbiology 2011, 2. 76. Weinstein DL, O'Neill BL, Hone DM, Metcalf ES: Differential Early Interactions between Salmonella enterica Serovar Typhi and Two Other PathogenicSalmonella Serovars with Intestinal Epithelial Cells. INFECTION AND IMMUNITY 1998, 66(5):2310-2318. 77. Lambeth LS, Yao Y, Smith LP, Zhao Y, Nair V: MicroRNAs 221 and 222 target p27Kip1 in Marek's disease virus-transformed tumour cell line MSB-1. The Journal of general virology 2009, 90(Pt 5):1164-1171. 78. Orecchini E, Doria M, Michienzi A, Giuliani E, Vassena L, Ciafre SA, Farace MG, Galardi S: The HIV-1 Tat protein modulates CD4 expression in human T cells through the induction of miR-222. RNA Biology 2014, 11(4):334-338. 79. Pandis I, Ospelt C, Karagianni N, Denis MC, Reczko M, Camps C, Hatzigeorgiou AG, Ragoussis J, Gay S, Kollias G: Identification of microRNA-221/222 and microRNA-323-3p association with rheumatoid arthritis via predictions using the human tumour necrosis factor transgenic mouse model. Annals of the rheumatic diseases 2012, 71(10):1716-1723. 80. Xu T, Huang C, Chen Z, Li J: MicroRNA-323-3p: a new biomarker and potential therapeutic target for rheumatoid arthritis. Rheumatology International 2014, 34(5):721-722. 81. Qiu Y, Luo X, Kan T, Zhang Y, Yu W, Wei Y, Shen N, Yi B, Jiang X: TGF-beta upregulates miR-182 expression to promote gallbladder cancer metastasis by targeting CADM1. Molecular bioSystems 2014, 10(3):679-685. 82. Kelada S, Sethupathy P, Okoye IS, Kistasis E, Czieso S, White SD, Chou D, Martens C, Ricklefs SM, Virtaneva K et al: miR-182 and miR-10a are key regulators of Treg specialisation and stability during Schistosome and Leishmania-associated inflammation. PLoS Pathog 2013, 9(6):e1003451. 83. Zheng Y, Yin L, Chen H, Yang S, Pan C, Lu S, Miao M, Jiao B: miR-376a suppresses proliferation and induces apoptosis in hepatocellular carcinoma. FEBS Letters 2012, 586(16):2396-2403. 84. Formosa A, Markert EK, Lena AM, Italiano D, Finazzi-Agro E, Levine AJ, Bernardini S, Garabadgiu AV, Melino G, Candi E: MicroRNAs, miR-154, miR-299-5p, miR-376a, miR-376c, miR-377, miR-381, miR-487b, miR-485-3p, miR-495 and miR-654-3p, mapped to the 14q32.31 locus, regulate proliferation, apoptosis, migration and invasion in metastatic prostate cancer cells. Oncogene 2013. 85. Nachmani D, Zimmermann A, Oiknine Djian E, Weisblum Y, Livneh Y, Khanh Le VT, Galun E, Horejsi V, Isakov O, Shomron N et al: MicroRNA editing facilitates immune elimination of HCMV infected cells. PLoS Pathog 2014, 10(2):e1003963. 86. Ogoshi K, Hashimoto S-i, Nakatani Y, Qu W, Oshima K, Tokunaga K, Sugano S, Hattori M, Morishita S, Matsushima K: Genome-wide profiling of DNA methylation in human cancer cells. Genomics 2011, 98(4):280-287. 87. Alleaume C, Eychene A, Caigneaux E, Muller J-M, Philippe M: Vasoactive intestinal peptide stimulates proliferation in HT29 human colonic adenocarcinoma cells: concomitant activation of Ras/Rap1–B-Raf–ERK signalling pathway. Neuropeptides 2003, 37(2):98-104. 88. Postigo AA: Opposing functions of ZEB proteins in the regulation of the TGFbeta/BMP signaling pathway. The EMBO journal 2003, 15:2443-2452. 89. Skeen VR, Collard TJ, Southern SL, Greenhough A, Hague A, Townsend PA, Paraskeva C, Williams AC: BAG-1 suppresses expression of the key regulatory cytokine transforming growth factor beta (TGF-beta1) in colorectal tumour cells. Oncogene 2013, 32(38):4490-4499. 90. Bachman KE, Blair BG, Brenner K, Bardelli A, Arena S, Zhou S, Hicks J, Marzo AD, Argani P, Park BH: p21(WAF1/CIP1) mediates the growth response to TGF-beta in human epithelial cells. Cancer biology and therapy 2004, 3(2):221-225. 91. Matlashewski G, Banks L, Pim D, Crawford L: Analysis of human p53 proteins and mRNA levels in normal and transformed cells. European Journal of Biochemistry 1986, 154(3):665-672. 92. MEYER DH, K. P, SREENIVASAN, M. P: Evidence for Invasion of a Human Oral Cell Line by Actinobacillus actinomycetemcomitans. INFECTION AND IMMUNITY 1991:2719-2726. 93. Fink SL, Cookson BT: Pyroptosis and host cell death responses during Salmonella infection. Cellular microbiology 2007, 9(11):2562-2570. 94. Xiao G: Autophagy and NF-kappaB: Figureht for fate. Cytokine & growth factor reviews 2007, 18(3-4):233-243. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/57573 | - |
dc.description.abstract | 沙門氏菌(Salmonella)是一種可以在細胞內生長的革蘭氏陰性菌,沙門氏菌會藉由糞口傳染且可以感染人和動物,是目前造成食物中毒主要的病原菌之一。沙門氏菌會感染吞噬細胞與非吞噬細胞並在宿主細胞內進行複製,當宿主細胞被沙門氏菌感染時,細胞會啟動初級免疫反應以對抗沙門氏菌的感染。已發現微核醣核酸(microRNAs)會參與宿主細胞抵抗病原菌的防禦,目前已有許多研究證實微核醣核酸可以調控初級免疫反應、發炎反應以及抗微生物感染的免疫反應。然而,微核醣核酸在胞內細菌感染中所扮演的角色仍未清楚。在我們的研究當中,我們利用沙門氏菌感染人類大腸上皮細胞HT-29後,發現宿主細胞的miR-146a表現量會增加。為進一步探討宿主miR-146a在沙門氏菌感染時所扮演的角色,我們使用沙門氏菌感染大量表現miR-146a的HT-29細胞株,再利用菌落形成單位(colony formation unit)分析細胞內細菌數以觀察細菌在細胞內生長的情形。結果發現在大量表現miR-146a的HT-29細胞株中的細菌數比對照組有明顯減少。為了更進一步探討miR-146a是否具有抑制細胞內細菌的能力,我們另外選用HCT116和HeLa兩種細胞株,進行沙門氏菌感染並觀察其是否具有相同現象。首先我們利用MTT證實miR-146a在細胞內大量表現後並不會影響細胞的生長速度。接著再利用先前實驗方法感染沙門氏菌並進行分析,從實驗結果中發現當細胞有表現miR-146a時,細胞內的細菌數的確會比對照組少,而在細胞數目上則沒有明顯的差異。此外我們也使用高通量細胞影像分析系統(high content analysis)作為連續觀察細胞內沙門氏菌生長情形的影像工具,以HT-29和HeLa細胞進行觀察,此影像系統分析的結果與菌落形成單位的結果相符合。 | zh_TW |
dc.description.abstract | Salmonella, a Gram negative intracellular bacterium is an important and widespread pathogen causing foodborne infections in both human beings and animals. Salmonella can infect phagocytic and non-phagocytic cells, surviving and replicating within host cells. The innate immunity of host cells is triggered to against Salmonella infection. Regarding to the host responses against pathogens, it has been reported that microRNAs (miRNAs) are involved in this process, and miRNAs have been identified as an important regulator of innate immunity, inflammatory response and anti-microbial immunity. However, the role of miRNA in host response to intracellular bacterial infection is still largely unclear.
In our study, we found that miR-146a was induced in HT-29 cells after Salmonella infection. To understand the role of miR-146a in Salmonella infection, Salmonella was used to infect miR-146a-overexpressed HT-29 cells, and the colony formation unit (CFU) assay was used to measure the intracellular bacterial numbers after Salmonella infection. The results showed that the intracellular CFU was significantly decreased in miR-146a-transfected cells compared with the mock cells. To determine whether the inhibitory activity of miR-146a is a universal phenomenon, we measured the impact of miR-146a on two other Salmonella-infected epithelial cells, HCT116 and HeLa cells, and found that miR-146a did not affect cell viability of HCT116 and HeLa cells assayed by MTT assays. After Salmonella infection, the results showed that the intracellular CFU was decreased in miR-146a expressed cells compared with the mock cells. Furthermore, we also used a high-throughput microscopy, high content analysis (HCA), as an image tool to investigate the intracellular growth of Salmonella continuously in both HT-29 cells and HeLa cells, demonstrating that the data were consistent with the CFU assay. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T06:52:09Z (GMT). No. of bitstreams: 1 ntu-103-R01424007-1.pdf: 4111816 bytes, checksum: bae58e2c16bc681c28b1fc167e92e4d9 (MD5) Previous issue date: 2014 | en |
dc.description.tableofcontents | 口試委員會審定書 I
Acknowledgement II 中文摘要 III Abstract IV 1. Introduction 1 1.1. Introduction of Salmonella spp. 2 1.1.1. Pathogenesis of Salmonella 2 1.2. Host responses to enteric bacterial infection 4 1.2.1. Epithelial integrity and epithelial cell turnover 5 1.2.2. Innate immune responses 5 1.2.3. Autophagy 6 1.3. MicroRNAs 7 1.3.1. MicroRNAs against pathogen infection 8 1.4. Aim of this study 11 2. Materials and Methods 13 2.1. Bacteria strains and culture conditions 14 2.2. Cell culture and transfection 14 2.3. In vitro infection assay 15 2.4. Enumeration of intracellular bacteria 16 2.5. Cell viability assay 16 2.6. RNA isolation 17 2.7. Reverse transcription for miRNAs 18 2.8. Quantification of miRNAs by qRT-PCR 18 2.9. Construction of miRNAs expressing plasmids 19 2.10. Establish miR-146a mix clone in HCT116 cells and HeLa cells 19 2.11. MTT cell viability assay 20 2.12. Image-based high content assay 21 2.13. Statistical analysis 23 3. Results 24 3.1. The optimal condition for Salmonella infection in HT-29 cells 25 3.2. The impacts of identified host miRNAs on intracellular replication of Salmonella 26 3.3. Establishment mixed stable clone in other epithelial cells 27 3.4. Time course of intracellular CFU in HCT116 and HeLa cells infected with Salmonella 29 3.5. The expression of miR-146a in HCT116 and HeLa cells limits intracellular replication of Salmonella 29 3.6. Imaged-based high content assay for monitoring the intracellular replication of Salmonella 31 4. Discussion 33 5. Figures 42 Figure 1. Optimization of the multiplicity of infection. 44 Figure 2. Overexpression of microRNAs in HT-29 cells. 46 Figure 3. The transient overexpression of host miRNAs may restrict Salmonella intracellular replication 49 Figure 4. The transient overexpression of host miR-146a, miR-182* and miR-376a limits Salmonella intracellular replication 50 Figure 5. Expression of miR-146a in HCT116 and HeLa epithelial cells 53 Figure 6. Time course of intracellular CFU in HT-29 infected with Salmonella. 54 Figure 7. The miR-146a inhibits Salmonella intracellular replication in epithelial HCT116 and HeLa cells. 55 Figure 8. Monitoring in vitro Salmonella infection assay by high content analysis 60 6. References 64 | |
dc.language.iso | en | |
dc.title | 微核醣核酸146a於沙門氏菌感染宿主細胞之角色探討 | zh_TW |
dc.title | Role of Host microRNA-146a in Salmonella Infection | en |
dc.type | Thesis | |
dc.date.schoolyear | 102-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 賴信志,楊翠青,顏伯勳,邱浩傑 | |
dc.subject.keyword | 沙門氏菌,微核醣核酸,微核醣核酸146a,胞內繁殖,病原體-宿主交互作用,高通量細胞影像分析系統, | zh_TW |
dc.subject.keyword | Salmonella,miRNAs,miR-146a,intracellular replication,pathogen-host interaction,high content analysis, | en |
dc.relation.page | 79 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2014-07-22 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 醫學檢驗暨生物技術學研究所 | zh_TW |
顯示於系所單位: | 醫學檢驗暨生物技術學系 |
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ntu-103-1.pdf 目前未授權公開取用 | 4.02 MB | Adobe PDF |
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