Journal: Medicine
Article Title: Curcumin induces endoplasmic reticulum stress-associated apoptosis in human papillary thyroid carcinoma BCPAP cells via disruption of intracellular calcium homeostasis
doi: 10.1097/MD.0000000000011095
Figure Lengend Snippet: Curcumin induces phosphorylation of IRE1α and XBP-1 mRNA splicing. BCPAP cells were exposed to different dosages (12.5–50 μM) of curcumin for 24 hours. After the cells were collected, western blot or RT-PCR analysis were performed. (A) Curcumin increases the phosphorylation of IRE1α in BCPAP cells. The protein levels of phosphorylated IRE1α and total IRE1α were detected by western blot analysis. β-actin was used as a loading control. (B) Curcumin increases XBP-1 splicing in BCPAP cells. The mRNA levels of spliced and unspliced forms of XBP-1 were assessed by RT-PCR. Actin was performed as a loading control. (C) Curcumin treatment results in the conversion of inactive unspliced XBP-1 (XBP-1u) protein to an active spliced (XBP-1s) protein in BCPAP cells. The protein levels of spliced and unspliced forms of XBP-1 were detected by western blot assay. (D) Curcumin enhances the mRNA expressions of XBP-1 downstream genes. BCPAP cells were exposed to different dosages (12.5–50 μM) of curcumin for 24 hours. The mRNA expressions of ERDJ3 , EDEM1 , and SERP1 were determined by RT-PCR and normalized to that of Actin . The data shown represent the means ± S.D. of 3 independent experiments. && P < .01, # P < .05, ∗ P < .05, ∗∗ P < .01 versus SC group. (E) 4-PBA fails to rescue the XBP-1 splicing induced by curcumin in BCPAP cells. Cells were pretreated with or without different dosages of 4-PBA (2.5–10 mM) for 1 hour. After that, cells were treated with 50 μM of curcumin for 24 hours and XBP-1 splicing was measured by RT-PCR. IRE1α = inositol-requiring enzyme 1α, RT-PCR = reverse transcriptase-polymerase chain reaction.
Article Snippet: The antibodies used were as follows: anti-ATF6 (sc-22799), anti-XBP-1 (sc-7160), anti-CaMKII (sc-9035), anti-CHOP (sc-7351), anti-PARP (sc-7150), anti-β-actin (sc-47778), and goat anti-mouse (sc-2005), or rabbit (sc-2004) IgG-HRP were purchased from Santa Cruz Biotechnology (Santa Cruz, CA).
Techniques: Western Blot, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction
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