Recombinant lentivirus targeting the pleotrophin gene reduces pleotrophin protein expression in pancreatic cancer cells and inhibits neurite outgrowth of dorsal root ganglion neurons

Yao,Jun; Li,Wen-Yao; Li,Shuo-Guo; Feng,Xiao-Shan; Gao,She-Gan

doi:10.3892/mmr.2014.1918

Recombinant lentivirus targeting the pleotrophin gene reduces pleotrophin protein expression in pancreatic cancer cells and inhibits neurite outgrowth of dorsal root ganglion neurons

Authors:
- Jun Yao
- Wen-Yao Li
- Shuo-Guo Li
- Xiao-Shan Feng
- She-Gan Gao
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Affiliations: Department of Oncology, First Affiliated Hospital, Henan University of Science and Technology, Luoyang, Henan 471003, P.R. China
Published online on: January 24, 2014 https://doi.org/10.3892/mmr.2014.1918
Pages: 999-1004

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Abstract

The objectives of the present study were to construct the recombinant primate lentivirus‑short hairpin RNA-pleiotrophin (pLV-shRNA-PTN) vector, to investigate the silencing effect of pLV-shRNA-PTN on PTN expression in MIA PaCa-2 cells and to observe the inhibition efficiency of pLV-shRNA‑PTN on neurite outgrowth from dorsal root ganglion (DRG) neurons in vitro. The construction procedure for recombinant lentivirus pLV-shRNA-PTN has been described previously. In the present study, pLV-shRNA‑PTN was used to infect MIA PaCa-2 pancreatic cancer cells and the efficiency of the knockdown of the PTN gene on day 7 following infection was analyzed using western blotting. The morphological changes in the cultured DRG neurons were observed by monoculture of DRG neurons and co-culture with MIA PaCa-2 cells in vitro. The recombinant lentivirus pLV-shRNA‑PTN was successfully constructed. The western blot analysis showed that the inhibition rates of PTN expression were 46, 80, 20 and 21%, respectively, following pLV-shRNA‑PTN-A, B, C and D infection. pLV-shRNA-PTN‑B showed the highest knockdown efficiency. DRG neurons co-cultured with infected MIA PaCa-2 cells were decreased in size when compared with the control, and there was a significant decrease in the number and length of neurites. The results suggest that efficient and specific knockdown of PTN in MIA PaCa-2 pancreatic cancer cells and the subsequent reduction in PTN expression results in the inhibition of neurite outgrowth from DRG neurons.

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