摘要
目的:检测在化疗药引起胃癌细胞DNA损伤过程中端粒酶、TRF1和TRF2的表达.方法:用不同浓度足叶乙甙处理胃癌细胞SGC7901和MKN28,分别在3,6,12,24和36h进行检测.采用实时定量TRAP分析检测端粒酶活性;用实时RT-PCR检测hTERTmRNA表达;用Westernblot和实时RT-PCR检测TRF1和TRF2表达.结果:两种细胞端粒酶活性及TRF2mRNA表达均在药物处理的早期明显增高(P<0.05),且显示明显的药物浓度依赖性(P<0.05);TRF1表达稍增高,但无统计学意义(P>0.05).hTERTmRNA表达无明显改变(P>0.05).TRF2表达在蛋白和mRNA水平均增高(P<0.05).结论:端粒酶和TRF2参与化疗药引起的胃癌细胞DNA损伤反应.
AIM： To investigate the possible involvement of telomerase and telomeric repeat binding factors （TRF1 and TRF2） in chemotherapeutic agentsinduced DNA damage responses in gastric cancer cells.
METHODS： Gastric cancer cell line SGC7901 and MKN28 were treated with various concentrations of etoposide for 3, 6, 12, 24 and 36 h. Telomerase activity was measured by realtime quantitative telomeric repeat amplification protocol （RTQ-TRAP） assay. The expression of human telomerase reverse transcriptase （hTERT） mRNA was detected by real time reverse transcription polymerase chain reaction （RT-PCR）. The expression of TRF1 and TRF2 were detected by Western blot and real time RT-PCR at protein and mRNA level, respectively.
RESULTS： Telomerase activity and TRF2 mRNA expression were up-regulated at the early stage of drug treatment in both cell lines （P 〈 0.05）. The expression of TRF1 mRNA was also increased, but it was not significant （P 〉 0.05）. The increase of telomerase activity was independent on hTERT mRNA levels, and TRF2 was significantly increased both in protein and mRNA levels （P 〈 0.05）. The up-regulation was in a drug dose-dependent manner.
CONCLUSION： Telomerase activity and TRF2 expression is possibly involved in the responses of gastric cancer cells to DNA-damaging drugs.
出处
《世界华人消化杂志》
CAS
北大核心
2006年第10期942-946,共5页
World Chinese Journal of Digestology
基金
国家自然科学基金资助项目
No.30400016~~
