Title page for etd-0019115-165758


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URN etd-0019115-165758
Author Jun-Yan Zeng
Author's Email Address No Public.
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Department Biological Sciences
Year 2014
Semester 1
Degree Master
Type of Document
Language zh-TW.Big5 Chinese
Title New Combretastatin A-4 enediyne derivatives induce Program
Cell Death in Neuroblastoma SH-SY5Y cell line
Date of Defense 2015-01-07
Page Count 69
Keyword
  • Combretastatin A-4
  • LO-OMe
  • LO-NH2
  • PARP
  • Caspase-3
  • Apoptosis
  • Autophagy
  • p62
  • LC3
  • Abstract Antimitotic drug, combretastatin A-4 (CA-4) is a natural plant extract exhibits powerful anticancer and anti-angiogenesis activity by targeting microtubule formation. CA-4 enediyne derivatives, such as LO-OMe and LO-NH2 were shown to possess potent cytotoxicity in various cancer cell lines except human neuroblastoma cell. Here, we further investigate the cytotoxic effects of LO-OMe and LO-NH2 on neuroblastoma SH-SY5Y cells. Our viability assay demonstrated that LO-OMe and LO-NH2 mediated 50 percent killing of SH-SY5Y cells at a concentration of 3.2 and 2.1 ┬ÁM, respectively. Cell lysates of drug treated cells were collected for analyzing markers of cellular autophagy and apoptosis using antibodies against LC3, p62, caspase-3, cleaved PARP. The results indicated a dose-dependent increase of LC3-II/LC3-I ratio and p62 protein upon drug treatments. In addition, consistent effects were observed in GFP-LC3 reporter H4 cell line. Both compounds elicited the increase in the number and the size of GFP-LC3 puncta, indicating an accumulation of autophagosomes in reporter cells. Our previous data showed LO-OMe and LO-NH2 treatment led to cell cycle arrest at G2/M. In addition, LO-OMe and LO-NH2 also significantly unregulated the level of active caspase-3, which resolved in the increase of cleaved-PARP. In summary, our results showed that LO-OMe and LO-NH2 induced the increase of autophagosomes. The accumulation of p62 indicated an inhibition in the fusion between autophagosome and lysosome, which might be due to the interference of microtubule polymerization exerted by these compounds. Unresolved cellular stress then resulted in programmed cell death of SH-SY5Y neuroblastoma cells. However, the molecular details awaits further investigation.
    Advisory Committee
  • Ming-Jung Wu - chair
  • Hong Yi-Ren - co-chair
  • Jiin-Tsuey Cheng - advisor
  • Files
  • etd-0019115-165758.pdf
  • Indicate in-campus at 5 year and off-campus access at 5 year.
    Date of Submission 2015-01-23

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