Down-regulation of Sonic hedgehog signaling pathway activity is involved in 5-fluorouracil-induced apoptosis and motility inhibition in Hep3B cells

  1. Qiyu Wang1,†,
  2. Shuhong Huang1,‡,
  3. Ling Yang1,
  4. Ling Zhao2,
  5. Yuxia Yin2,
  6. Zhongzhen Liu1,
  7. Zheyu Chen2,*,
  8. Hongwei Zhang1,*

Article first published online: 29 SEP 2008

DOI: 10.1111/j.1745-7270.2008.00456.x

Issue

Acta Biochimica et Biophysica Sinica

Acta Biochimica et Biophysica Sinica

Volume 40, Issue 9, pages 819–829, September 2008

Additional Information

How to Cite

Wang, Q., Huang, S., Yang, L., Zhao, L., Yin, Y., Liu, Z., Chen, Z. and Zhang, H. (2008), Down-regulation of Sonic hedgehog signaling pathway activity is involved in 5-fluorouracil-induced apoptosis and motility inhibition in Hep3B cells. Acta Biochimica et Biophysica Sinica, 40: 819–829. doi: 10.1111/j.1745-7270.2008.00456.x

Author Information

  1. 1

    School of Life Sciences, Shandong University, Jinan 250100, China

  2. 2

    School of Medicine, Shandong University, Jinan 250012, China

  1. †

    This work was supported by the grants from the National Natural Science Foundation of China (30570967, 30570967, 30228031, 30671072, 30671050 and 30725020) and the National Ministry of Science and Technology of China (2006CB503803, 2007CB947100 and 2007CB815800)

  2. ‡

    These authors contributed equally to this work

* *Corresponding authors: Hongwei Zhang: Tel, 86-531-88364935; Fax, 86-531-88565610; E-mail, zhw@sdu.edu.cn Zheyu Chen: Tel, 86-531-88382329; Fax, 86-531-88382329; E-mail, zheyuchen@sdu.edu.en

Publication History

  1. Issue published online: 29 SEP 2008
  2. Article first published online: 29 SEP 2008
  3. Received: March 25, 2008; Accepted: June 26, 2008

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Keywords:

  • Sonic hedgehog signaling pathway;
  • hepatocellular carcinoma;
  • 5-fluorouracil;
  • cell apoptosis;
  • cell motility

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.

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