Open AccessDown‐regulation of Sonic hedgehog signaling pathway activity is involved in 5‐fluorouracil‐induced apoptosis and motility inhibition in Hep3B cells
Author(s) -
Wang Qiyu,
Huang Shuhong,
Yang Ling,
Zhao Ling,
Yin Yuxia,
Liu Zhongzhen,
Chen Zheyu,
Zhang Hongwei
Publication year - 2008
Publication title -
acta biochimica et biophysica sinica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.771
H-Index - 57
eISSN - 1745-7270
pISSN - 1672-9145
DOI - 10.1111/j.1745-7270.2008.00456.x
Subject(s) - gli1 , sonic hedgehog , hedgehog signaling pathway , biology , signal transduction , viability assay , microbiology and biotechnology , gene knockdown , apoptosis , motility , cell growth , cyclopamine , cancer research , biochemistry
The Sonic hedgehog (SHh) pathway plays a critical role in normal embryogenesis and carcinogenesis, but its function in cancer cells treated with 5‐fluorouracil (5‐FU) remains unknown. We examined the expression of a subset of SHh signaling pathway genes, including SHh, SMO, PTC1, Su(Fu) and HIP in human hepatocellular carcinoma (HCC) cell lines, Hep3BandHepG2, treated with 5‐FU by reverse transcription‐polymerase chain reaction. Using trypan blue analysis, 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazoliumbromide assay and terminal deoxynucleotidyl transferase‐mediated digoxigenin‐dUTP nick‐end labeling assay, we also detected the apoptosis of Hep3B cells resulting from the transfection of pCS2‐Glil expression vectorcombined with 5‐FU treatment. The motility of the cells was detected by scratch wound closure assay. The expression and subcellular location of PTC1 protein in Hep3B cells treated by 5‐FU were also investigated by Western blot analysis and immunofluorescent microscopy. The results indicated that the expression of SHh pathway target molecules at both messenger RNA and protein levels are evidently down‐regulated in Hep3B cells treated with 5‐FU. The overexpression of GUI restores cell viability and, to some extent, the migration abilities inhibited by 5‐FU. Furthermore, 5‐FU treatment affects the subcellular localization of PTC1 protein, a key member in SHh signaling pathway. Our data showed that the down‐regulation of SHh signaling pathway activity was involved in 5‐FU‐induced apoptosis and the inhibition of motility in hedgehog‐activated HCC cell lines. This implies that the combination of SHh signaling pathway inhibitor and 5‐FU‐based chemotherapy might represent a more promising strategy against HCC.