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The Effects of ER Stress on Cell Toxicity and Htt Levels in the Stable PC12 Cells Expressing Htt586

Author: WuZuo
Tutor: QinZhengHong；LinFang；GuZhenLun
School: Suzhou University
Course: Pharmacology
Keywords: Huntington’s disease Huntingtin (Htt) Endoplasmic Reticulum stress (ER stress) autophagy/lysosome ubiquitin-proteasome
CLC: R363
Type: Master's thesis
Year: 2011
Downloads: 17
Quote: 0
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Abstract


Objective: To study the activation of Endoplasmic Reticulum stress (ER stress) induced by tunicamycin on the metabolism of Htt-586 in PC12 cells expressing wild-type Htt586 or mutant Htt586.Methods: Cellular models of Huntington’s disease was established using the PC12 cells stably expressing a caspase 6-cleavage product Htt-586 with 18Q (wild-type) or 100Q (mutant) using lentivirus. The effects of different concentrations of tunicarmycin (TM) on cell survival were assessed with MTT method; The levels of CHOP, Caspase-12,Bcl-2, LC3II,P62 and Htt586 were determined by Western blot analysis;Results: Wild-type and mutant Htt-586 were stably expressed in highly differentiated PC12 cells with lentivirus transfection. Endoplasmic reticulum stress (ER stress) of PC12 cells can be induced after cell treated with TM (250μg/L) for 36 hr, ER stress related protein CHOP and caspsase-12; autophagy related protein LC3-II and p62 were increased. Meanwhile, Bcl-2 was reduced. TM treatment increased mutant Htt586 levels.Conclusion: In this study, we have successfully established stable PC12 cellular models of Huntington’s disease using lentivirus-mediated expression of a caspase-6 cleavage fragment Htt-586 with 18Q (wild type) or 100Q (mutant). ER stress could be induced by tunicamycin in these stable cell lines. ER stress increased accumulation of mutant Htt586.

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