Bufalin Induces Mitochondria-Dependent Apoptosis in Pancreatic and Oral Cancer Cells by Downregulating hTERT Expression via Activation of the JNK/p38 Pathway

Figure 3

hTERT silencing triggers mitochondria-dependent apoptosis of CAPAN-2 and CAL-27 cells. (a–c) CAPAN-2 and CAL-27 cells were transfected with scrambled siRNA, hTERT siRNA1, or hTERT siRNA2 for 48 h. The nontransfected cells were included for comparison. (a) The hTERT mRNA expression determined by qRT-PCR. Data were normalized to β-actin and expressed as fold change relative to the control. (b) The hTERT protein expression determined by western blot. Data were normalized to GAPDH. (c) The protein levels of procaspase-9/-3 and cleaved caspase-9/-3 determined by western blot. Data were normalized to β-actin. versus control. (d–f) CAPAN-2 and CAL-27 cells transfected with scrambled siRNA, hTERT siRNA1, or hTERT siRNA2 were treated with 100 nM bufalin or vehicle alone for 24 h. The nontransfected cells were included for comparison. (d) Cell viability determined by the MTT assay. (e) The telomerase activity determined by the telomerase PCR enzyme-linked immunosorbent assay. (f) DNA damage determined by the comet assay. The comet tail length was indicative of the extent of DNA damage. versus control with the corresponding DMSO or bufalin treatment. , .

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