n-Butylidenephthalide induced apoptosis in the A549 human lung adenocarcinoma cell line by coupled down-regulation of AP-2alpha and telomerase activity.
OBJECTIVE
To investigate the role of hTERT gene expression and AP-2alpha in n-butylidenephthalide (n-BP)-induced apoptosis in A549 lung cancer cells.


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Wei
Chyou-wei
CW
Institute of Biomedical Nutrition, College of Medicines and Nursings, Hungkuang University, Taichung, Taiwan, China;
Lin
Chai-ching
CC
Yu
Yung-luen
YL
Lin
Chai-yi
CY
Lin
Po-cheng
PC
Wu
Min-tze
MT
Chen
Cheng-jueng
CJ
Chang
Wenliang
W
Lin
Shinn-zong
SZ
Chen
Yi-lin Sophia
YL
Harn
Horng-jyh
HJ
D000230:Adenocarcinoma / Q000201:enzymology / Q000235:genetics*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D015536:Down-Regulation / Q000187:drug effects; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects* / Q000235:genetics; D006801:Humans; D008175:Lung Neoplasms / Q000201:enzymology / Q000235:genetics*; D051379:Mice; D010796:Phthalic Anhydrides / Q000494:pharmacology*; D012334:RNA, Neoplasm / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019098:Telomerase / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D050656:Transcription Factor AP-2 / Q000187:drug effects*; D014407:Tumor Cells, Cultured; D023041:Xenograft Model Antitumor Assays
D010796:Phthalic Anhydrides; D012334:RNA, Neoplasm; D050656:Transcription Factor AP-2; C509186:TERT protein, human; D019098:Telomerase; C026105:butylidenephthalide
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