Study On The Effects Of Inhibiting HTERT Expression And Paclitaxel On Proliferation And Apoptosis In Hut78 Cells

Objectives1. To investigate the effects of telomerase antisense oligodeoxynucleotide (ASODN) on telomerase activity and cell proliferation in cutaneous T-cell lymphoma (CTCL) cell Hut78 line.2. To observe the effects of apoptosis induction of paclitaxel on Hut78 cells line in vitro, and to explore the mechanism of antitumour of paclitaxel.3. To explore whether simultaneous shortening of telomeres and inhibition of telomerase may provide combined antitumor activity and whether telomerase inhibitors can be used as a sensitizing agent to increase the cytotoxic effect of subtoxic concentrations of paclitaxel in Hut78 cells.4. To provide an easy, quick and economical method to synthesize siRNA with T7 RNA polymerase in vitro transcription, and the reactive condition is further optimized. To study the effect of hTERT-siRNA on telomerase activity and cell proliferation in Hut78 cells.Methods1. Design and synthesize oligonucleotides directed against hTERT and transfect them into Hut78 cellsAccording to the hTERT cDNA, we designed hTERT antisense oligonucleotide to the region complementary to the beginning code region as well as sense oligonucleotide, and all were protected by phosphorothioate bonds. All oligonucleotides sequences were subjected to a BLAST search of the National Center for Biotechnology Information expressed sequence tag library to ensure that they targeted only the desired gene. They were introduced into Hut78 cells by lipofectamine-mediated DNA transfection technique.