OBJECTIVE@#To express matrix
remodeling associated 7 (MXRA7) in the
human acute myeloid leukemia SHI-1
cell line and to assess the
role of MXRA7 in the
biological function of SHI-1
cells.@*
METHODS@#The full-length
cDNA sequence of
human MXRA7 was synthesized and subcloned into the
lentivirus shuttle vector pRRL-
Venus. SHI-1
cells were transfected with the
lentivirus which was packaged with
293T cells. The YFP-positive
cells were sorted by
flow cytometry and the stable
cell lines were obtained by expanded
culture. The expression and distribution of MXRA7 in SHI-1
cells were verified by real-
time qPCR,
Western blot and
laser confocal
techniques.
Cell proliferation and
cell cycle were measured by
flow cytometry, and
apoptosis was determined by
Annexin V and 7-AAD
staining. The expression of
apoptosis related
proteins were detected by
Western blot.@*RESULTS@#The stable SHI-1
cell line overexpressing MXRA7 was established successfully.
Laser confocal
analysis confirmed that MXRA7 was expressed in the
cytoplasm of SHI-1
cells. Compared with the control
cell line, the overexpression of MXRA7 showed no effect on the
cell proliferation and
cell cycle, but reduced the percentage of
apoptosis cells induced by
methotrexate. Moreover, the expression of BCL-2
protein was increased by overexpression of MXRA7, which can inhibit
cell apoptosis.@*CONCLUSION@#The SHI-1 stable
cell line overexpressing MXRA7 was established successfully, and MXRA7 could inhibit
drug-induced
apoptosis through increasing the expression of BCL-2
protein.