Inhibitory effects of antisense phosphorothioate oligodeoxynucleotides on pancreatic cancer cell Bxpc-3 telomerase activity and cell growth in vitro.
OBJECTIVE
To investigate the effect of telomerase hTERT gene antisense oligonucleotide (hTERT-ASO) on proliferation and telomerase activity of pancreatic cancer cell line Bxpc-3.


METHODS
MTT assay was used to detect the effect of different doses of hTERT-ASO on proliferation of Bxpc-3 cell for different times. To study the anti-tumor activity, the cells were divided into three groups: Control group (pancreatic cancer cell Bxpc-3); antisense oligonucleotide (hTERT-ASO) group; and nosense oligonucleotide group decorated with phosphorothioate. Telomerase activity was detected using TRAP-PCR-ELISA. Cell DNA distribution was examined using flow cytometry assay. Cell apoptosis was observed by transmission electron microscope in each group.


RESULTS
After treatment with 6 mmol/L hTERT-ASO, cell proliferation was inhibited in dose- and time-dependent manner. The telomerase activity decreased after treatment with hTERT-ASO for 72 h. Flow cytometry showed the cell number of G0/G1 phase increased from 2.7% to 14.7%, the cell number of S phase decreased from 72.7% to 51.0%, and a sub-G1 stage cell apoptosis peak appeared in front of G1 stage.


CONCLUSIONS
Telomerase antisense oligodeoxy-nucleotide can inhibit the proliferation of pancreatic cancer cell line Bxpc-3 and decrease the telomerase activity and increase cell apoptosis rate in vitro.
Wang
Yun-Feng
YF
Department of General Surgery, The Center Hospital of Shanghai Yangpu District (Branch Hospital of the Affiliated Xinhua Hospital of Shanghai Jiaotong University), Shanghai 200127, China. wangyunfeng197911@hotmail.com
Guo
Ke-Jian
KJ
Huang
Bei-Ting
BT
Liu
Yong
Y
Tang
Xiao-Yun
XY
Zhang
Jian-Jun
JJ
Xia
Qiang
Q
D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism*; D015316:Genetic Therapy; D006801:Humans; D020319:Oligodeoxyribonucleotides, Antisense / Q000494:pharmacology*; D010190:Pancreatic Neoplasms / Q000201:enzymology / Q000473:pathology*; D019098:Telomerase / Q000235:genetics* / Q000378:metabolism*
D004268:DNA-Binding Proteins; D020319:Oligodeoxyribonucleotides, Antisense; D019098:Telomerase