Dissertation > Excellent graduate degree dissertation topics show

Promotion Effect of Hydrogen Sulfide on Tolerability of HepG-2Cells to ADM and the Underlying Mechanisms

Author: YinXiaoGang
Tutor: QinChunHong
School: Nanhua University
Course: Department of General Surgery
Keywords: Hydrogen sulfide HepG-2 multidrug resistance KATPchannel BDNF-TrkBpathway
CLC: R735.7
Type: Master's thesis
Year: 2013
Downloads: 1
Quote: 0
Read: Download Dissertation

Abstract


【Objective】The hepatocellular carcinoma cell lines HepG-2treating with adriamycin（ADM） was deserved as experimental model （ADM-killing HepG-2model）,and then the role of hydrogen sulfide （H2S） in tolerance of HepG2to ADMand its mechanism were investigated on the basis of experimental model.【Methods】1. HepG-2cell were cultured in vitro, treated with ADM and NaHS. TheIC50value of ADM was analyzed by CCK8assay. In view of ADM IC50,experiment was divided into four groups, containing control, ADM,ADM+NaHS, NaHS group. HepG-2apoptosis was detected by propidiumiodide （PI） staining and flow cytometry （FCM）. Meanwhile, expression levelof caspase-3protein was determined by ELISA. Western blot was used todetermine expression of multidrug resistance （MDR） protein Bcl-2,P-glycoprotein （P-gp）. To known about whether H2S promoted tolerance ofHepG-2to ADM.2. HepG-2cell were cultured in vitro, treated with ADM, NaHS,glybenclamide (Gly, inhibitor of KATPchannel). The IC50value of ADM wasanalyzed by CCK8assay. HepG-2apoptosis was detected by propidiumiodide （PI） staining and flow cytometry （FCM） in five group, containingcontrol, ADM, ADM+NaHS, ADM+NaHS+Gly, Gly group. Western blot was used to determine expression of Bcl-2and P-glycoprotein （P-gp） in four group,containing control, NaHS, NaHS+Gly, Gly. To known about whether KATPchannel mediated the role of H2S promoting tolerance of HepG-2to ADM.3. According to ADM IC50, experiment was divided into five groups,containing control, ADM, ADM+NaHS, ADM+NaHS+K252a （inhibitor ofBDNF-TrKB pathway）, K252group. Apoptosis and cell viability of the fivegroups were determined by CCK8and FCM, respectively. Western blot wasused to determine expression of Bcl-2, P-glycoprotein （P-gp） and BDNF infour groups, containing control, NaHS, NaHS+K252a, K252a. To known aboutwhether BDNF-TrkB pathway mediated the role of H2S promoting toleranceof HepG-2to ADM.【Results】1. The effects of H2S on tolerance of HepG-2to ADM1.1IC50of ADMIC50of ADM in NaHS （100、200、400μmol/L） treatment group were0.59±0.05mg/L,0.78±0.07mg/L,1.42±0.08mg/L, higher than control group（only treatment with ADM,0.32±0.03mg/L）（P<0.01, P<0.001）.1.2ApoptosisApoptosis rate of ADM group （77.9±0.9）%was higher than that of controlgroup （16.9±0.6）%（P<0.001）. Comparing with ADM group （77.9±0.9）%,apoptosis rate of ADM+NaHS group （25.7±2.3）%significantly decreased（P<0.001）. Apoptosis rate of NaHS group was lower than that of controlgroup （16.9±0.6）%（P<0.01）.1.3Caspase-3expressionExpression of caspase-3in ADM group （1.09±0.05ng/ml） was higher thanthat of control group （0.53±0.03ng/ml）（P<0.001）. Comparing with ADMgroup （1.09±0.05ng/ml）, expression of caspase-3in ADM+NaHS group（0.81±0.03ng/ml） significantly decreased （P<0.01）. There was no significantdifference between control group and NaHS group （P＞0.05）. 1.4Bcl-2and P-gp expressionExpression of Bcl-2in NaHS（100、200、400μmol/L）group was higher thanthat of control group （P<0.05, P<0.001）. Expression of P-gp in NaHS（200、400μmol/L）group was higher than that of control group （P<0.01, P<0.001）.2. The mediating role of KATPchannel for H2S promoting tolerance ofHepG-2to ADM2.1IC50of ADMIC50of ADM in NaHS group （0.78±0.07mg/L） was higher than that ofADM group （0.32±0.03mg/L）（P<0.001）. IC50of ADM in NaHS+Gly group（0.58±0.01mg/L） was lower than that of NaHS group （0.78±0.07mg/L）（P<0.05）.2.2ApoptosisApoptosis rate of ADM group （77.9±0.9）%was higher than that of controlgroup （16.9±0.6）%（P<0.001）. Comparing with ADM group （77.9±0.9）%,apoptosis rate of ADM+NaHS group （25.7±2.3）%significantly decreased（P<0.001）. Apoptosis rate of ADM+NaHS+Gly group[（50.5±3.9）%] was higherthan that of ADM+NaHS group （25.7±2.3）%（P<0.01）. There was no significantdifference in apoptosis rate between control group and Gly group（P＞0.05）.2.3Bcl-2and P-gp expressionExpression of Bcl-2and P-gp protein in NaHS group was higher than thatof control group（P<0.05, P<0.01）. Expression of Bcl-2and P-gp protein inNaHS+Gly group was lower than that of NaHS group（P<0.05）. There was nosignificant difference in expression of Bcl-2and P-gp protein between controlgroup and Gly group（P＞0.05）.3. The mediating role of BDNF-TrkB pathway for H2S promoting toleranceof HepG-2to ADM3.1BDNF expressionExpression of BDNF in NaHS （100,200,400μmol/L） group was higherthan that of control group （P<0.05, P<0.001）.3.2Cell viability Cell viability of ADM group （OD value=0.56±0.02） was lower than thatof control group（1.90±0.03）（P<0.001）. Cell viability of ADM+NaHS group（1.39±0.09） was higher than that of ADM group （0.56±0.02）（P<0.001）. Cellviability of ADM+NaHS+K252a group （0.71±0.01） was lower than that ofADM+NaHS group （P<0.001）. There was no significant difference in cellviability between control group and K252group （P＞0.05）.3.3ApoptosisApoptosis rate of ADM group[（77.9±0.9）%] was higher than that ofcontrol group [（16.9±0.6）%]（P<0.001）. Apoptosis rate of ADM+NaSH group[（25.7±2.3）%] was lower than that of ADM group （77.9±0.9）%（P<0.001）.Apoptosis rate of ADM+NaHS+K252a group [（46.0±5.9）%] was higher thanthat of ADM+NaHS group [（25.7±2.3）%]（P<0.05）. There was no significantdifference in apoptosis rate between K252a group and control group（P＞0.05）.3.4Bcl-2and P-gp expressionExpression of Bcl-2and P-gp protein in NaHS group was higher than thatof control group（P<0.05，P<0.001）. Expression of Bcl-2and P-gp protein inNaHS+K252a group was lower than that of NaHS group（P<0.05，P<0.01）.There was no significant difference in expression of Bcl-2and P-gp proteinbetween K252group and control group（P＞0.05）.【Conclusion】1. H2S promoted the tolerance of HepG-2cell to ADM.2. KATPchannel and BDNF-TrkB pathway probably mediated the action ofH2S promoting tolerance of HepG-2cell to ADM.3. The action of promoting tolerance of HepG-2cell to ADM was associatedwith KATPchannel and BDNF-TrkB pathway.

Related Dissertations

  1. Screen Genes Affected the Activity of Quorum Sensing Regulator VqmA and Functional Analysis of LysR and MFS Proteins in V.choleare,R378
  2. Effects of Elemene on Topoisomerase Ⅰ Andⅱof Human Hepatocarcinoma HepG-2 Cells,R735.7
  3. Effects of SILA409 on the Expression of GRP78 in Nci-h446 Human Small Cell Lung Cancer Cells and Its Correlation with Chemotherapy Resistance to VP-16,R734.2
  4. Effect of GCS on Expression of MDR1mRNA and P-gp Function in K562/AO2 Cell Line,R733.7
  5. Establishment of Cisplatin- Resistant Gastric Cancer Cell SGC7901/DDP and the Effect of Recombinant Adenovirus Mediated Human IL-24 Gene to Chemosensitivity,R735.2
  6. Jieduhuayu party of the leukemia in K562/A02 HL60/ADR cell resistance reversal role,R733.7
  7. Expression and Significance of Breast Cancer Resistance Protein and Topoisomerase Ⅱ in Nude Mice Transplanted Tumor Model of A549/CDDP Multidrug Resistance Cell,R730.5
  8. Invitro and in vivo inhibitioon of MRP Gene Expression and Reversal of Multidrug Resistance by SIRNA,R73-3
  9. The Effect and Ion Channel Mechanism of Hydrogen Sulfide on Rat Heart Function,R363
  10. Hydrogen Sulfide Attenuates Abdominal Aortic Coarctation Induced Cardiac Hypertrophy and Fibrosis in Rats,R541
  11. Construction of the Fusion Protein Ag85B-Hsp16.3 of Mycobacterium Tuberculosis and Its Effects on Human Hepatoma Carcinoma Cell HepG-2,R735.7
  12. Xiaoaiping injection and oxaliplatin combined with inhibition of proliferation and induction of apoptosis in hepatoma cell research,R285
  13. Studies of Isomycin Derivant on Inducing Apoptsis of Human Hepatoma HepG-2 Cell Line,R285
  14. In Vitro Effects of the Proliferation and Apoptosis for Human Hepatoma Cell Line HepG-2 by Antisense Oligodeoxynucleotides Against Livin,R735.7
  15. The Synthesis and Apoptosis-inducing Effects and Mechanism on HepG-2 Cells of Flavone Derivatives,R965
  16. The oligomycin multidrug resistant cell line the K562/A02 role of research,R733.7
  17. P-glycoprotein inhibitors from traditional Chinese medicine in a high -throughput screening,R287
  18. Cardioprotective Effect of Hydrogen Sulfide to Heart Failure of Rats after Myocaidial Infarction and Its Possible Mechenism,R542.22
  19. Protection of Hydrogen Sulfide Delayed Preconditioning on Myocardial Ischemia-reperfusion Injury in Rats,R541
  20. Limited space confined workplaces hydrogen sulfide poisoning prevention measures,R136
  21. Hydrogen sulfide stress conditions regulate seed germination and seedling growth signaling mechanism,S565.1

CLC: > Medicine, health > Oncology > Gastrointestinal Cancer > Liver tumors
© 2012 www.DissertationTopic.Net  Mobile