[RNAi-mediated Silencing of CXCR4 Inhibits the Adhesion, Invasion and Tumorigenicity of Acute Monocytic Leukemic Cell Line SHI-1]

Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2015 Oct;23(5):1286-91. doi: 10.7534/j.issn.1009-2137.2015.05.012.
[Article in Chinese]

Abstract

Objective: To investigate the effect of CXCR4 gene on the proliferation, adhesion, invasion and tumorigenicity of a human monocytic leukemic cell line SHI-1.

Methods: The lentivirus vector silencing the expression of CXCR4 was constructed for infection of SHI-1 cells silencing expression of CXCR4 in SHI-1 cells. The expression of CXCR4, MMP-2 and MMP-9 was detected by real time PCR. The expression of CXCR4 on membrane of SHI-1 cells was detected by flow cytometry. The SHI-1 cell proliferation ability was detected by MTT. The co-culture system of the leukemia cells and bone marrow stromal cells was used to detect the adhesion and migration ability of leukemia cells. SHI-1 cells were inoculated subcutaneously in nude mice to investigate the growth ability in vivo.

Results: After SHI-1 cells were infected by lentivirus silencing expression of CXCR4, the expression of CXCR4 mRNA in SHI-1 CXCR-4i cells decreased by 76% as compared with expression of SHI-1/NC of negative control virus, the expression of CXCR4 on membrane of SHI-1/CXCR4i obviously downregulated; the expression of MMP-2 and MMP-9 in SHI-1/CXCRi cells also declined by 63% and 62% respectively; the proliferation ability of SHI-1/CXCR4i in vitro did not obviously changed, but the adhesion and trans-matrigel invasion ability significantly decreased, the SHI-1/CXCR4i cells could not form neoplasm subcutaneously in mice, but the SHI-1 and SHI-1/NC cells could form neoplasm subcutaneously in mice, and there was no significant difference in volumn of neoplasm mass.

Conclusion: The silencing expression of CXCR4 can decline the adhesion and migration ability of SHI-1 cells, and can completely suppress the formation of neoplasm subcutaneously, so the CXCR4 may serve as a target for treating leukemia.

MeSH terms

  • Animals
  • Cell Adhesion
  • Cell Line, Tumor
  • Cell Proliferation
  • Coculture Techniques
  • Humans
  • Lentivirus
  • Leukemia, Monocytic, Acute / pathology*
  • Matrix Metalloproteinase 2 / metabolism
  • Matrix Metalloproteinase 9 / metabolism
  • Mesenchymal Stem Cells
  • Mice
  • Mice, Nude
  • Neoplasm Invasiveness
  • RNA Interference*
  • RNA, Messenger
  • Receptors, CXCR4 / genetics*
  • Signal Transduction

Substances

  • CXCR4 protein, human
  • RNA, Messenger
  • Receptors, CXCR4
  • MMP2 protein, human
  • Matrix Metalloproteinase 2
  • MMP9 protein, human
  • Matrix Metalloproteinase 9