Can inhibition of telomerase increase pancreatic cancer cell's susceptibility to chemotherapeutic reagents?
OBJECTIVE
To clarify the inhibition of pancreatic cancer cells by interference with the hTR component of the telomerase reverse transcriptase enzymatic complex and evaluate susceptibility of antisense hTR pancreatic cancer cells to chemotherapeutic reagents.


METHODS
A 593 bp of full length hTR cDNA was subcloned into a mammalian expression vector pcDNA3.1(-) in antisense orientation to construct an antisense hTR expression plasmid. The plasmids were introduced into Pancl cells, a human pancreatic carcinoma cell line, by lipofectin, and G418-resistant stable transformants were expanded. Resulting stable clones were screened for the presence of hTR insert by PCR with T7 and BGH reverse primers located on the flanks of the multiclonal site of pcDNA3.1 vector. Cell growth rate, hTR expression, telomerase activity, and anchorage-independent growth property were analyzed. Finally, susceptibility of antisense hTR cells to chemotherapeutic reagents was evaluated.


RESULTS
Significant downregulation of endogenous hTR was evident in the antisense-hTR transformed cells, and telomerase activity was markedly decreased compared to control cells in standard TRAP assays. Furthermore, the proliferation and the anchorage-independent growth ability in antisense-hTR expressing cells were significantly decreased compared with the control parental cells. However, no crisis or senescence phenomena was observed. Antisense hTR appears to increase Pancl cell's susceptibility to chemotherapeutic reagent cDDP, but not to differentiation reagent DMSO, COX2 inhibitor sulinbac, NS-398, curcumin, and chemotherapeutic reagent adriamycin (ADM).


CONCLUSIONS
These data indicate that hTR is probably a critical component of human telomerase activity and that downregulation of the RNA component of human telomerase is an effective target for anticancer strategy and antisense hTR can increase Pancl cell's susceptibility to cDDP.
Teng
Li-Song
LS
Department of Oncological Surgery, Zhejiang University School of Medicine, Hangzhou 310003, China. lsteng@mail.hz.zj.cn
Fahey
Thomas J
TJ
D000230:Adenocarcinoma; D000970:Antineoplastic Agents / Q000494:pharmacology*; D002455:Cell Division / Q000187:drug effects; D045744:Cell Line, Tumor / Q000166:cytology / Q000187:drug effects; D016373:DNA, Antisense; D019008:Drug Resistance, Neoplasm; D006801:Humans; D010190:Pancreatic Neoplasms; D019098:Telomerase / Q000037:antagonists & inhibitors*
D000970:Antineoplastic Agents; D016373:DNA, Antisense; D019098:Telomerase