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[Effects of Down-Regulation of on Differentiation and Apoptosis of MPN Cells with Gene Mutation and Survival of 6133/MPL Mice]. | LitMetric

[Effects of Down-Regulation of on Differentiation and Apoptosis of MPN Cells with Gene Mutation and Survival of 6133/MPL Mice].

Zhongguo Shi Yan Xue Ye Xue Za Zhi

Blood Diseases Institute, Xuzhou Medical University; Department of Hematology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou 221000, Jiangsu Province, China.

Published: October 2024

AI Article Synopsis

  • The study investigates how reducing the protein PAK1 affects the growth, differentiation, and death of myeloproliferative neoplasm cells with a specific MPL mutation.
  • Researchers used various methods, including lentivirus-mediated shRNA technology and assays, to measure cell proliferation, apoptosis, and protein expression in these cells.
  • Results showed that decreasing PAK1 significantly hampers cell growth, increases polyploid DNA formation and apoptosis in 6133/MPL cells, and improves survival rates in affected mice.*

Article Abstract

Objective: To investigate the effects of down-regulation of p21 activated kinase 1 (PAK1) on the proliferation, differentiation, and apoptosis of myeloproliferative neoplasm (MPN) cells (6133/MPL) with thrombopoietin receptor MPL mutation at codon 515 () and survival of 6133/MPL mice.

Methods: Interference with the protein level of in 6133/MPL cells was assessed using lentivirus-mediated shRNA transfection technology. CCK-8 assay was used to detect the effect of down-regulation of on the proliferation viability of 6133/MPL cells, and colony-forming ability was measured by cell counting. Flow cytometry was used to detect the kinase activity on the ability of polyploid DNA formation and cell apoptosis in 6133/MPL cells. The expression of cyclin D1, cyclin D3 and apoptosis-related protein Bax was detected by Western blot. The infiltration of tumor cells in spleen and bone marrow of 6133/MPL mice were detected by HE staining.

Results: Down-regulation of inhibited the proliferation and reduced the ability of cell colony formation of 6133/MPL cells. After knocking down , the content of polyploid DNA in 6133/MPL cells increased from 31.8 to 57.5% and 48.0%, and the proportion of apoptosis increased approximately to 10.8%. Down-regulation of led to a reduction of infiltration of tumor cells in liver and bone marrow of 6133/MPL mice, thereby prolonging survival time.

Conclusion: Down-regulation of can significantly inhibit the growth of 6133/MPL cells, promote the formation of polyploid DNA, induce 6133/MPL cell apoptosis, and prolong the survival time of 6133/MPL mice.

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Source
http://dx.doi.org/10.19746/j.cnki.issn.1009-2137.2024.05.026DOI Listing

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