Title page for etd-0803114-151657


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URN etd-0803114-151657
Author Po-min Chang
Author's Email Address No Public.
Statistics This thesis had been viewed 5388 times. Download 738 times.
Department Biological Sciences
Year 2013
Semester 2
Degree Master
Type of Document
Language English
Title Effect of Methoxychlor on Ca2+ Homeostasis and Apoptosis in HA59T Human Hepatoma Cells
Date of Defense 2014-07-08
Page Count 36
Keyword
  • methoxychlor
  • apoptosis
  • Ca2+
  • hepatoma
  • Abstract Methoxychlor is an organochlorine pesticide. It has been shown that methoxychlor is thought to be an endocrine disrupter that affects Ca2+ homeostasis and cell viability in different cell models. However, the effect of methoxychlor on human hepatoma cell is unclear. This study explored the action of methoxychlor on cytosolic free Ca2+ concentrations ([Ca2+]i) and apoptosis in HA59T human hepatoma cells. Fura-2, a Ca2+-sensitive fluorescent dye, was applied to measure [Ca2+]i. Viability was measured by using the probe WST-1. Apoptosis was measured by Alexa Fluor® 488 Annexin V/propidium iodide (PI) staining. Methoxychlor at concentrations of 0.1-1 µM caused a [Ca2+]i rise in a concentration-dependent manner. Removal of external Ca2+ abolished methoxychlor’s effect. Methoxychlor-induced Ca2+ influx was confirmed by Mn2+-induced quench of fura-2 fluorescence. Methoxychlor-induced Ca2+ entry was inhibited by nifedipine, econazole, SKF96365, and protein kinase C modulators. Methoxychlor killed cells at concentrations of 10-130 µM in a concentration-dependent fashion. Chelation of cytosolic Ca2+ with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N' -tetraacetic acid/AM (BAPTA/AM) did not prevent methoxychlor’s cytotoxicity. Furthermore, methoxychlor (10 and 50 µM) induced apoptosis concentration- dependently. Together, in HA59T cells, methoxychlor induced a [Ca2+]i rise by inducing Ca2+ entry via protein kinase C-sensitive Ca2+-permeable channels, without causing Ca2+ release from stores. Methoxychlor also induced apoptosis that was independent of [Ca2+]i rises.
    Advisory Committee
  • Ko-L Lin - chair
  • Hsueh-Wen Chang - co-chair
  • Chao-neng Tseng - co-chair
  • Chung-Lung Cho - advisor
  • Chung-Ren Jan - advisor
  • Files
  • etd-0803114-151657.pdf
  • Indicate in-campus at 1 year and off-campus access at 1 year.
    Date of Submission 2014-09-07

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