Location of NLS-RARα protein in NB4 cell and nude mice

Abstract

In the majority of acute promyelocytic leukemia (APL) cases, translocons produce a promyelocytic leukemia protein-retinoic acid receptor α (PML-RARα) fusion gene. Studies have reported that neutrophil elastase (NE) cleaves bcr-1-derived PML-RAα in early myeloid cells, leaving only the nuclear localization signal (NLS) of PML attached to RARα. NLS-RARα promotes cell growth and inhibits differentiation in response to ATRA. However, the mechanisms by which NLS-RARα affects cell biological characteristics are yet to be fully elucidated. The present study found that the location of RARαwas altered after it was cleaved by NE. Firstly, NE was overexpressed during the preparation of recombinant plasmid NB-4/pCMV6-NE-Myc to cleave PML-RARα. The total protein expression levels of myc and NE and expression levels of NLS-RARα in nucleoprotein were detected by western blotting. Location of NLS-RARα protein was detected by immunofluorescence and confocal laser scanning. Secondly, a nude mice model was constructed and NE protein, NLS-RARα and RARα protein assays, and the location of NLS-RARα and RARα proteins were assessed as described. The present results showed that, compared with the control groups, the location of NLS-RARα protein was predominantly detected in the nucleus, whereas RARα was mainly distributed in the cytoplasm. These findings were consistent with those of the nude mice model, and these may be used as a foundation to explain the occurrence mechanism of APL.

RANBP9 suppresses tumor proliferation in colorectal cancer

“…2018-A001-01; Shanghai, China). All mice were handled according to the National Institutes of Health Guidelines for the Care and Use of Laboratory Animals (16). Three animals per group (4 groups: The control and RANBP9 -shRNA group for HCT116 cells or HT29 cells) were used in each experiment.…”

Tissue expression of retinoic acid receptor alpha and CRABP2 in metastatic nephroblastomas

“…RARA can be translocated from the cytoplasm to the nucleus after being synthesized, modified and stimulated by RA. Under physiological conditions, RARA is located in the cytoplasm and nucleus [21]. RARA gene mutations observed in promyelocytic leukemia can induce the cytoplasmic translocation of the receptor [22].…”

A novel retinoic acid analog, 4-Amino-2-Trifluoromethyl-Phenyl Retinate (ATPR) triggers differentiation and is effective in the treatment of Acute Promyelocytic Leukemia

“…It is not like solid tumor to establish tumor-bearing animal model by the xenotransplantation, hematological malignancies with dispersivity and generalized characteristics are difficult to propagate [11][12][13]. In recent years, a lot of human malignant hematopoietic cells, such as acute lymphoblastic leukemia (ALL), chronic myeloid leukemia (CML), and multiple myeloma (MM), have been successfully engrafted in nude mice [5,14].…”