Knockdown of hTERT by SiRNA suppresses growth of Capan-2 human pancreatic cancer cell via the inhibition of expressions of Bcl-2 and COX-2.
OBJECTIVE
To investigate the effects of human telomerase reverse transcriptase (hTERT) on the growth of Capan-2 human pancreatic cancer cell and apoptosis.


METHODS
Cell proliferation, apoptosis and cell cycle were analyzed by cell counting and flow cytometry. mRNA and protein expressions of hTERT, Bcl-2 and cyclooxygenase (COX)-2 were assessed by real time PCR and Western blot.


RESULTS
Cell growth was significantly inhibited by 26.39 percent 24 h after hTERT-small interference RNA (siRNA) transfection (50 nmol/L) with a 100 percent silencing efficiency (P < 0.05). The inhibition rates of cell proliferation were 46.77 percent, 70.61 percent, 84.71 percent and 85.99 percent at 2, 3, 5 and 7 days after transfection, respectively (P < 0.001). Early and late apoptotic cells increased significantly (especially 24 h after transfection) (P < 0.001). The cell cycle was suppressed significantly as manifested by the increase of cells in the G0/G1 phase and the decrease of cells in the S phase and G2/M phase (P < 0.01). The expressions of Bcl-2 mRNA and COX-2 mRNA were inhibited significantly 48 h after transfection: the inhibition rates were 86.86 percent and 100 percent, respectively (P < 0.001). Levels of Bcl-2 protein were downregulated by 58.54 percent and 63.44 percent and the levels of COX-2 protein were downregulated by 50.06 percent and 82.77 percent at 48 h and 72 h after transfection, respectively.


CONCLUSIONS
Knockdown of hTERT by siRNA can inhibit the growth of Capan-2 cell. The inhibitory effect is associated with the downregulation of Bcl-2 and COX-2.
Zhong
Ying Qiang
YQ
Department of Gastroenterology, The Second Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong Province, China. zhongyingq@yahoo.com.cn
Xia
Zhong Sheng
ZS
Fu
Yu Ru
YR
Zhu
Zhao Hua
ZH
D017209:Apoptosis / Q000502:physiology; D002453:Cell Cycle / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D045744:Cell Line, Tumor; D051546:Cyclooxygenase 2 / Q000235:genetics*; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D010190:Pancreatic Neoplasms / Q000235:genetics* / Q000473:pathology / Q000628:therapy*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics*; D034741:RNA, Small Interfering; D019098:Telomerase / Q000235:genetics*
D019253:Proto-Oncogene Proteins c-bcl-2; D034741:RNA, Small Interfering; D051546:Cyclooxygenase 2; C509186:TERT protein, human; D019098:Telomerase