Algariri, Eman Salem Saeed
(2022)
Gene Expression, Biochemical And
Functional Analysis Of Stromal
Interaction Molecule 1 (Stim1) Silencing
In Acute Myeloid Leukemia Cell Lines.
Masters thesis, Universiti Sains Malaysia.
Abstract
A stromal interaction molecule (STIM1) was recently discovered to be a critical
modulator of cell growth and survival in a variety of cancers. In contrast, the function of
STIM1 in AML is still not properly understood. Therefore, the aim of the present study is
to investigate the effect of STIM1 on the proliferation and survival of AML cell lines by
employing a dicer-substrate siRNA (dsiRNA)-mediated silencing technique performed on
two AML cell lines, namely THP-1 and Kasumi-1 cells. The expression profile of critical
genes involved in proliferation, survival, and ROS production was analyzed through RTqPCR
after STIM1 silencing. The intracellular calcium and ROS levels, cell proliferation,
and colony formation were also measured after silencing of STIM1. The study findings
revealed that STIM1 mRNA expression was higher in THP-1 cells compared to Kasumi-
1 cells. STIM1 silencing, at the mRNA and protein levels, was achieved in THP-1 cells
using 10 nM dsiSTIM1 for 24 hours and 20 nM dsiSTIM1 for 48 hours in Kasumi-1 cells.
STIM1 silencing resulted in down-regulation of KRAS, MAPK, C-MYC, Akt, NOX2 and
PKC in both AML cells, whereas BAX was up-regulated, and Bcl-2 was down-regulated
only in THP-1 cells. Furthermore, STIM1 silencing resulted in a reduction in the
intracellular calcium and ROS levels and inhibition of AML cell proliferation and colony
formation in both AML cells. In conclusion, the present study suggests that STIM1 might
have a crucial role in promoting the proliferation and survival of AML cells
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