Dissertation > Excellent graduate degree dissertation topics show

RNAi-mediated inhibition of survivin CNE-2 nasopharyngeal carcinoma xenograft experimental study

Author: ZhaoLiuFang
Tutor: LiXiaoJiang
School: Kunming Medical College
Course: Oncology
Keywords: RNA interference Plasmid Survivin Short hairpin RNA survivin Nasopharyngeal carcinoma Gene Therapy Mice Bare
CLC: R739.63
Type: Master's thesis
Year: 2011
Downloads: 50
Quote: 1
Read: Download Dissertation

Abstract


The first part of the NPC survivinshRNA targeting plasmid expression vector construct pSIREN-survivin/shRNA identification purposes recombinant plasmid and sequencing for exploring gene therapy for nasopharyngeal carcinoma RNA interference (RNAi) basis. Materials and methods based on gene pool survivin cDNA sequence (NM 0 01168) and Reynolds design principles, design and synthesis of restriction sites at both ends of the 65-base oligonucleotide strands, annealing into complementary duplexes later T4DNA enzyme was cloned into the linearized RNAi-Ready pSIREN-DNR-DsRed-Express plasmid; transformed into E. coli DH-5a strain, plasmid line restriction enzyme digestion and sequencing analysis. Results PCR amplified fragment with the expected results; double digestion confirmed recombinant vector was successfully constructed, insert fragment sequencing results with synthetic oligonucleotide sequences consistent. Conclusion The recombinant plasmid pSIREN-survivin/shRNA, for the next turn with liposome package and nasopharyngeal carcinoma cell research foundation. The second part of the small interfering RNA inhibition of survivin gene expression induced apoptosis in nasopharyngeal carcinoma CNE-2 Objective To investigate the application of small interfering RNA (small interfering RNA, siRNA) technology nasopharyngeal carcinoma CNE-2 cells inhibited survivin expression on cells wither death and proliferation. Materials and methods design, synthesis specifically inhibits the expression of survivin shRNA, transfected CNE-2 cells, semi-quantitative RT-PCR analysis of transfected cells survivin mRNA changes in flow cytometry cell cycle and apoptosis, MTT test cell proliferation. Results plasmid transfection efficiency was approximately (46.9 ± 1.55)%; positive transfected plasmid 24h survivinmRNA and protein inhibition rates were: (41.58 ± 0.63)%, (68.29 ± 0.52)%; 48h inhibition rates were increased (63.64 ± 0.96)%, (70.83 ± 0.48)%; positive plasmid group 24h apoptosis rate was (36.24 ± 0.78)%, 48h increased to (50.37 ± 0.85)%, significantly higher than the negative control group and the untreated group, positive plasmid group S phase cells decreased, G2 / M phase increased the proportion of the emergence of G2 / M phase arrest phenomena; MTT showed positive plasmid transfected cell growth slowed. Conclusion specific shRNA can effectively interfere with nasopharyngeal carcinoma cells survivin mRNA and protein expression and induce apoptosis and inhibit cell proliferation. The third part of the RNAi-mediated inhibition of survivin CNE-2 nasopharyngeal carcinoma xenograft experiments targeting survivin gene Objective To investigate the short hairpin RNA (short hairpin RNA, shRNA) in human nasopharyngeal carcinoma CNE2 tumor growth in nude mice affected. Materials and Methods Construction tumor model in nude mice by intratumoral multi-point injection, liposome transfection plasmid volumetric measurement of tumor growth, by semi-quantitative RT-PCR and immunohistochemical determination of survivin mRNA and protein expression, with a transmission electron microscope detect survivin shRNA on apoptosis and liver and kidney function were detected in nude mice. Results survivin gene sequence-specific shRNA can efficiently and stably suppress nasopharyngeal carcinoma xenografts survivin mRNA and protein expression; significantly inhibited tumor growth, promote apoptosis, and the mice had no effect on short-term kidney function. Conclusion RNAi technology can effectively inhibit the expression of survivin nasopharyngeal carcinoma xenograft reduce nasopharyngeal carcinoma cell proliferation and promote apoptosis of nasopharyngeal carcinoma cells, slowing tumor growth.

Related Dissertations

  1. The Expression of Let-7 microRNA during the Process of the Mouse Embryonic Lung Development and the Construction of the Adenovirus Shuttle Plasmid,Q78
  2. Study of Sepia Ink-Astragalus Preparation for Relieving Damage Caused by Chemotherapy,R285.5
  3. Polyethylenimine Conjugated Stearic Acid-g-chitosan Oligosaccharide Micelles for Antitumor Gene Therapy,R450
  4. Expression Dynamics of Pheromone Binding Proteins and Expression Influences by Mating and Knockdown of Cryl in Spodoptera Exigua,S433.4
  5. Molecular Cloning, Mrna Expression and Rnai of Nadph-Cytochrome P450 Reductase Gene in Helicoverpa Armigera (Hǘbner),S435.622.3
  6. Effects of Angiogenic Factor on Follicular Angiogenesis and Development in Immature Mouse,S865.13
  7. Study on Gene Expression of β-defensin1 in Mammary Gland of Mice,S852.4
  8. Roles of MMP-7 and Lysozyme in the Pathogenesis of Ulcerative Colitis Induced by DSS in Balb/c Mice,S858.91
  9. Some Pharmacology and Toxicity of Three Prescriptions from Chinese Veterinary Pharmacopoeia,S859.5
  10. Effects of BMPR-IB Gene Silencing by Small Interfering RNA on Apoptosis of Porcine Pollicular Granulosa Cells and Ecpression of BMP Pathy-Way-Ralted Genes,S828
  11. Inhibition of Myostatin (MSTN) Expresstion in Primary Porcine Fetal Fibroblasts by Lentivector-mediated RNAi,S828
  12. Study of Melittin on Tumor Inhibition and Part of Mechanism of Human Hepatocellular Carcinoma HepG-2 Xenograft in Nude Mice,R735.7
  13. Apolipoprotein E Gene Knock-out and High-fat Diet on IP3、 IP3R-1 ＆ Aβ Expression in Neurons of Mice’s Hippocampal CA1 ＆ Ca3,R329
  14. Effects of TACI-Ig on MRL/lpr Mice and Activation of JAK1-STAT1 Signaling Pathway in Lupus Nephritis,R363
  15. The Construction and Evaluation of a Novel Non-viral Gene Transfection System and Its Application in Mesenchymal Stem Cells Gene Recombination,R346
  16. Effects and Some Mechanisms of TACI-Ig on Immune Function in MRL/lpr Mice,R593.241
  17. Maternal Exposure to Fenvalerate in Embryo Accelerated Age-related Changes of Behaviors in CD-1 Mice,R714.2
  18. A Preliminary Study on the Regulation of Hepatoma Cell Biological Behavior by MCFP,R735.7
  19. The Experiment Study Was Used Three Methods to Repair the Ligation of the Femoral Nerve,R651.3
  20. The DNA Cloning and RNA Interference of Some Functional Genes in Laodelphax Striatellus (Fallen),S435.112.3
  21. Effect of β-elemene Combined with Radiotherapy on the Angiogenesis of Lung Adenocarcinoma Xenograft Tumor in Nude Mice,R734.2

CLC: > Medicine, health > Oncology > Department of Otolaryngology tumor > Pharyngeal tumors
© 2012 www.DissertationTopic.Net  Mobile