Liu Yong-ping, Hu Yue-di, Wang Feng, Ling Yang, Kong Ying-ze, Li Peng
Department of Oncology, Changzhou Tumor Hospital Affiliated to Medical School of Soochow University, Changzhou 213001, China.
Zhonghua Yi Xue Za Zhi. 2009 Sep 15;89(34):2391-4.
To evaluate whether antisense oligonucleotides (ASODN) targeting hTERT mRNA could sensitize human pancreatic cancer cell to gemcitabine in vitro.
hTERT mRNA expression was measured by real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) assay. Telomerase activity was examined by TRAP, polyacrylamide gel electrophoresis and silver staining. The proliferation capacity and the apoptosis of cancer cells were evaluated by MTT assay and double staining with both Annexin V-FITC and PI.
Transient transfection in human pancreatic cancer cell with hTERT ASODN diminished the abundance of hTERT mRNA in a concentration-dependent fashion. And 0.2 micromol/L ASODN decreased the telomerase activity by 0.47-fold in cancer cell. The IC(50) value of gemcitabine in ASODN-transfected cell was (0.8 +/- 0.2) micromol/L while that in oligofectamine-transfected control cell (7.3 +/- 0.9) micromol/L with a statistically significant difference. hTERT ASODN significantlly increased the gemcitabine-induced apoptosis rate in pancreatic cancer cell. The gemcitabine-induced apoptosis rate in ASODN-transfected cell was 60.28% while that in oligofectamine-transfected control cell 17.34%.
hTERT antisense oligodeoxynucleotide can increase the sensitivity of pancreatic cancer cell to gemcitabine. The mechanism may be due to the down-regulated expression of hTERT mRNA and telomerase activity.
评估靶向人端粒酶逆转录酶（hTERT）mRNA的反义寡核苷酸（ASODN）在体外是否能使人类胰腺癌细胞对吉西他滨敏感。
采用实时定量逆转录聚合酶链反应（RT-PCR）检测hTERT mRNA表达。通过端粒重复序列扩增法（TRAP）、聚丙烯酰胺凝胶电泳和银染检测端粒酶活性。采用MTT法及Annexin V-FITC和PI双染法评估癌细胞的增殖能力和凋亡情况。
用hTERT ASODN瞬时转染人胰腺癌细胞后，hTERT mRNA丰度呈浓度依赖性降低。0.2 μmol/L的ASODN可使癌细胞端粒酶活性降低0.47倍。ASODN转染细胞中吉西他滨的半数抑制浓度（IC50）值为（0.8±0.2）μmol/L，而在脂质体转染的对照细胞中为（7.3±0.9）μmol/L，差异有统计学意义。hTERT ASODN显著提高了吉西他滨诱导的胰腺癌细胞凋亡率。ASODN转染细胞中吉西他滨诱导的凋亡率为60.28%，而脂质体转染的对照细胞为17.34%。
hTERT反义寡脱氧核苷酸可提高胰腺癌细胞对吉西他滨的敏感性。其机制可能是由于hTERT mRNA表达下调和端粒酶活性降低。
