Dissertation > Excellent graduate degree dissertation topics show

Long-Fragment RNA Inhibits Hepatitis B Virus Gene Replication and Expression in HepG2.2.15 Cell Line

Author: TianLv
Tutor: HeSong
School: Chongqing Medical University
Course: Internal Medicine
Keywords: RNAi HBV replication antisense RNA
CLC: R512.62
Type: Master's thesis
Year: 2011
Downloads: 25
Quote: 0
Read: Download Dissertation

Abstract


Objective:To evaluate the inhibitory effects of long antisense RNA on HBV replication in HepG2.2.15 cells.Methods : The coding region of HBV S gene was cloned into pTARGET vector in sense and antisense orientations and the recombinant plasmids were transfected into HepG2.2.15 cells, and the HepG2.2.15 cells were divided into HBS2(antisense RNA) group,HBS4(sense RNA) group,interferon group, poly:c group and control group. HBsAg and HBeAg in the culture supernatant were detected by ELISA. HBV DNA,HBV cccDNA in the cells and HBV DNA in the supernatant were quantified by real time PCR.β-IFN mRNA in the cells was quantified by real time RT-PCR. The rate of apoptosis was detected by Cellular DNA Fragmentation ELISA.Results:(1)PCR and sequencing results displayed that recombinant plasmids were constructed successfully.(2)ELISA result showed that the level of HBsAg and HBeAg in the supernatant of HBS2 group, interferon group and poly:c group were lower than control group(P<0.05); the level of HBsAg in the supernatant of HBS4 group was higher than control group(P<0.05),but the level of HBeAg of HBS4 group had no difference with control group(P>0.05).(3)Real time PCR result displayed that the level of HBV DNA both in supernant and cells of HBS2 group, interferon group and poly:c group were lower than control group(P<0.05),but HBS4 group had no difference with control group(P>0.05),the level of HBV cccDNA in the cells of HBS2 group, interferon group and poly:c group were lower than control group(P<0.05);HBS4 group had no difference with control group(P>0.05).(4)Real time RT-PCR result indicated that the level ofβ-IFN mRNA in the cells of HBS2 group and poly:c group were higer than control group(P<0.05), HBS4 group had no difference with control group.(5) Cellular DNA Fragmentation ELISA result showed that the rates of apoptosis of HBS2 group, interferon group and poly:c group were higher than control group(P<0.05), HBS4 group had no difference with control group(P>0.05).Conclusion:Antisense RNA might be a useful tool to repress HBV replication,also could induce the apoptosis of HepG2.2.15 cells.

Related Dissertations

  1. Improvement of the Resistance of Nicotiana Tabaccum to Virus by RNAi and Initial Establishment of Genetic Transformation System of L.regale,S435.72
  2. Study on Roles of Diacylglycerol Kinase Gene Family in Rice under Xylanase and NaCl Treatments by Transient RNA Interference System,S511
  3. Construction and Identification of Recombinant Lentivirus Expressing shRNA Targeting to ORF1 and ORF7 of Porcine Reproductive and Respiratory Syndrome Virus Genomes,S852.65
  4. Cloning and Characterization of Prodh Gene cDNA from Leptinotarsa Decemlineata (Say) and Bacterial Expression of dsRNA,S435.32
  5. Toxicities of Several Newly-Typed Insecticides to Colorado Potato Beetle, Leptinotarsa Decemlineata (Say) and Molecular Cloning of Two Kinds of Targets,S435.32
  6. Expression of β-Catenin in Pig’s Ovary and the Effect of β-Catenin on Porcine Granulosa Cells Apotosis and Steroidogenesis Related Enzyme,S828
  7. RNAi transgenic silkworm BmNPV Resistance of inhibition,Q78
  8. Study on Genes Expression and Function of UDP-glucosyltransferases in the Silkworm, Bombyx Mori,S881.2
  9. Apoptosis Mechanisms Research in Interfering TIGAR Gene and Epirubicin Treatmnet in Hepatic Carcinoma Cells,R735.7
  10. Effects Suppression of Bcl-xl Expression by RNAi on Proliferation and Apoptosis of Human Pancreatic Cancer Cells and Chemosensitivity to Gemcitabine,R735.9
  11. The Clinical Significance of B7-H3 Expression in Human Prostate Cancer and Preliminary Research in Its Mechanism,R737.25
  12. Effects of Combined Therapy of 5-FU and Inhibitor of ClassⅢ　PI3K Human Gastric Cancer SGC7901 Cells,R735.2
  13. Effect of RNAi-mediated Gene Silencing of Mcl-1 on Apoptosis of Human Pancreatic Cancer Cell Line PANC-1,R735.9
  14. The Construction of pBI121-TPAS ihpRNA Vector of a Rice Tn Pong and Agrobacterium-mediated Transformation by Regeneration of Embryonic Callus,S511
  15. Silkworm BmMP54 polyhedrin gene and gene fusion expression and function of the preliminary study,Q78
  16. Study on a New Oncolytic Adenovirus for the Targeting Therapy to Malignancies,R730.5
  17. The Effect of Treatment from Dual Target in Murine Breast Tumors by Survivin and Vegf-c,R737.9
  18. Study of Apoptosis in MCF-7 and Its Mechanism Caused by VEGF siRNA Transfected into It,R346
  19. Silencing of the EGFR Gene by siRNA Effects on Sensitivity of Chemotherapy in MDA-MB-231 Cell Line,R737.9
  20. The Epression of Hypoxia-inducible Factor-1 Alpha in Cutaneous Squamous Cell Carcinoma and Its Role in Promoting Cell Proliferation,R739.5
  21. Cloning and Functional Analysis of MaChy Gene in Metarhizium Acridum,S476.12

CLC: > Medicine, health > Internal Medicine > Infectious disease > Viral infections > Viral Hepatitis > Hepatitis B
© 2012 www.DissertationTopic.Net  Mobile