The antibodies against p53, ANXA7, NF-[kappa]B p65, [beta]-actin, and horseradish peroxidase-conjugated secondary antibodies were from Santa Cruz Biotechnology (USA).
EECP Regulated the Levels of ANXA7, p53, and NF-[kappa]B p65 in MCF-7 and MDA-MB-231 Cells.
Our results showed that EECP potentially exerted its antitumor effect by inhibiting cell proliferation, inducing apoptosis, inhibiting cell migration, regulating ANXA7 and p53 levels, downregulating NF-[kappa]B p65 level and inhibiting its translocation from cytoplasm to nuclei, and increasing intracellular ROS level, decreasing mitochondrial membrane potential.
ANXA7, a member of the annexin family of calcium-dependent phospholipid binding proteins, codes for [Ca.sup.2+] dependent GTPase, which involves several different roles in autophagy, exocytosis, carcinogenesis, and tumor suppression [26-28].
In addition, potential roles of AR-A014418 in downregulation of splicing factors were reflected with decrease in Anxa7 (VA) variant and increase in Anxa7 WT tumor suppressor transcript and protein.
GSK3beta Inhibition Induced Reversal in Anxa7 Alternative Splicing.
Earlier we have shown that Anxa7 is a EGFR negative regulator; its allelic loss has been depicted with enhanced glioma survival signaling pathway [11, 13].
Here we have observed splicing deregulation with elevated Anxa7 VA (variant) transcript with loss in Anxa7 VB (WT) in comparison with Normal Brain cDNA Clontech company in U87 and U373 glioma cells (Figure 5(a)).
And a recent study indicated that annexin a7 (ANXA7), a member of the annexin family of calcium-dependent phospholipid binding proteins, was an endogenous regulator of PC-PLC.
Considering the important roles of ANXA7 and PC-PLC in atherosclerosis and propolis modulated atherosclerosis and affected PC-PLC activity, we hypothesized that propolis may also affect ANXA7, the endogenous regulator of PC-PLC.
Primary antibodies against ANXA7, NF-[kappa]B p65, GAPDH, and secondary antibody (horseradish peroxidase) were from Santa Cruz Biotechnology (USA).
Cells were incubated with ANXA7 and NF-[kappa]B p65 primary antibodies (1/100) at 4[degrees]C overnight.