Wei Chyou-wei, Lin Chai-ching, Yu Yung-luen, Lin Chai-yi, Lin Po-cheng, Wu Min-tze, Chen Cheng-jueng, Chang Wenliang, Lin Shinn-zong, Chen Yi-lin Sophia, Harn Horng-jyh
Institute of Biomedical Nutrition, College of Medicines and Nursings, Hungkuang University, Taichung, Taiwan, China;
Acta Pharmacol Sin. 2009 Sep;30(9):1297-306. doi: 10.1038/aps.2009.124. Epub 2009 Aug 24.
To investigate the role of hTERT gene expression and AP-2alpha in n-butylidenephthalide (n-BP)-induced apoptosis in A549 lung cancer cells.
Viability of A549 cells was measured by MTT assay. Protein expression was determined by Western blot. Telomerase activity was measured using the modified telomere repeat amplification protocol (TRAP) assay. Xenograft mice were used as a model system to study the cytotoxic effect of n-BP in vivo. The morphology of tumor was examined by immunohistochemical staining.
The growth of A549 lung cancer cells treated with n-BP was significantly inhibited. Telomerase activity and hTERT mRNA expression were determined by telomeric repeat amplification protocol and reverse transcription-polymerase chain reaction, respectively. n-BP inhibited telomerase activity and hTERT mRNA expression in A549 cells while overexpression of hTERT could abolish BP-induced growth inhibition in the A549 cells. We also showed that hTERT promoter activity in the presence of n-BP was mediated via AP-2alpha. We saw an inhibition of tumor growth when nude mice carrying A549 subcutaneous xenograft tumors were treated with n-BP. Immunohistochemistry of this tumor tissue also showed a decrease in the expression of hTERT.
The antiproliferative effects of n-BP on A549 cells in vitro and in vivo suggest a novel clinical application of this compound in the treatment of lung cancers.
探讨人端粒酶逆转录酶（hTERT）基因表达和AP - 2α在正丁烯基苯酞（n - BP）诱导A549肺癌细胞凋亡中的作用。
采用MTT法检测A549细胞活力。通过蛋白质免疫印迹法测定蛋白质表达。使用改良的端粒重复序列扩增法（TRAP）检测端粒酶活性。以异种移植小鼠作为模型系统研究n - BP在体内的细胞毒性作用。通过免疫组织化学染色检查肿瘤形态。
用n - BP处理的A549肺癌细胞生长受到显著抑制。分别通过端粒重复序列扩增法和逆转录 - 聚合酶链反应测定端粒酶活性和hTERT mRNA表达。n - BP抑制A549细胞中的端粒酶活性和hTERT mRNA表达，而hTERT的过表达可消除BP诱导的A549细胞生长抑制。我们还表明，在n - BP存在下hTERT启动子活性是通过AP - 2α介导的。当携带A549皮下异种移植肿瘤的裸鼠用n - BP处理时，我们观察到肿瘤生长受到抑制。该肿瘤组织的免疫组织化学也显示hTERT表达降低。
n - BP在体外和体内对A549细胞的抗增殖作用表明该化合物在肺癌治疗中具有新的临床应用价值。
