Fig. 2.

CDDO-Me inhibits hTERT gene and hTERT protein expression in pancreatic cancer cells. (A) Effect of CDDO-Me on hTERT gene expression. MiaPaCa-2 and Panc-1 cells were treated with CDDO-Me (0–5 μM) for 48 h and total cellular RNA was prepared using TRI-zole reagent. 1 μg of cellular RNA was reverse transcribed using oligo-dt primer and high fidelity reverse transcriptase. 1 μl of cDNA was amplified using hTERT or GAPDH primers. Amplified products were separated on 2% DNA agarose gel. Gels were stained with ethidium bromide and amplified DNA fragments were identified by base pair sizes. (B) Effect on hTERT protein. MiaPaCa-2 and Panc-1 cells were treated with CDDO-Me (0–5 μM) for 48 h and cell lysates were analyzed for hTERT and p-hTERT protein by Western blotting. Each experiment was repeated at least two times.