Zhongguo aizheng zazhi (Jun 2021)

The influence of PRUNE2 gene point mutation on proliferation, apoptosis, invasion and migration of prostate cancer DU145 cells

  • CAO Dalong, ZHU Wenkai, SHI Guohai, ZHANG Hailiang, WANG Ziliang, YE Dingwei

DOI
https://doi.org/10.19401/j.cnki.1007-3639.2021.06.001
Journal volume & issue
Vol. 31, no. 6
pp. 441 – 446

Abstract

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Background and purpose: PRUNE2 is a specific prognostic gene for neuroblastoma and plays an important role in regulating cellular differentiation, proliferation and invasion of neuroblastoma. Low expression of PRUNE2 gene is associated with poor prognosis of prostate cancer. The aim of this paper was to investigate the correlation of PRUNE2 mutation with proliferation, apoptosis, invasion and migration of prostate cancer DU145 cells. Methods: Recombinant plasmids carrying wild-type and mutant-type PRUNE2 were constructed and then transfected to prostate cancer DU145 cell line to construct corresponding stable transgenic strains. Cell counting kit-8 (CCK-8) assay and clone formation assay were used to detect the ability of cell proliferation. Apoptosis assay was used to detect the ability of apoptosis, and transwell assay was used to detect the abilities of invasion and migration of cells. Western blot was used to detect the level of protein expression. The interaction between proteins was detected by immunofluorescence and immunoprecipitation experiments. Results: DU145 cells transfected with mutant-type PRUNE2 E370K exhibited significantly increased proliferation, invasion and migration, but significantly decreased apoptosis (P<0.005). It was also found that PRUNE2 and RhoA could bind to each other, while the binding ability between mutant-type PRUNE2 E370K and RhoA was significantly weakened. We also found that mutant-type PRUNE2 E370K promoted the expressions of ROCK and FAK proteins related with Rho pathway and the expressions of Bcl-2 and cyclin D1 proteins related with cell proliferation, and inhibited the expression of E-cadherin associated with cell invasion and migration. Conclusion: PRUNE2 gene mutation positively promotes the proliferation, invasion and migration, and inhibits apoptosis of prostate cancer DU145 cells. These results provide important theoretical basis for studying the invasion and migration of prostate cancer.

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