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Advanced Neurology TG100-115 suppresses glioblastoma cell functions

Our study showed that TG100-115 attenuated PI3K/ invasion, and treatment resistance. Recently, Akt was
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Akt signaling. Akt, the central molecule of PI3K/Akt identified as a direct substrate of TG100-115, and kinase-
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signaling, undergoes activation by phosphorylation, deficient murine neutrophils confirmed the essential role
promoting tumor cell survival, proliferation, migration, of TRPM7 kinase in neutrophil function via Akt/mTOR
signaling. Our findings exhibited that TG100-115
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A
markedly reduced Akt phosphorylation. It should be noted
that TG100-115 was previously recognized as a potent
PI3K inhibitor, particularly targeting the PI3K-γ and δ
isoforms. 22,23,32 We performed MTT and wound-healing
assays using PIK-294, a selective blocker of PI3K p110δ.
A PI3K p110δ inhibitor was specifically used because
GBMs express a low level of PI3K p110γ. In Figure S4A,
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our data revealed that PIK-294 significantly reduced the
viability of U251 cells at higher doses (120 and 180 µM),
but not at lower doses (10, 30, and 60 µM), with 24-h
B
incubation. In contrast, TG100-115 significantly reduced
cell viability even at a lower dose of 30 µM after 24 h of
incubation. This data suggests that the PI3K inhibitory
activity of TG100-115 could partially affect the reduction
of cell viability.
Previous studies have indicated that PI3K p110-δ is
essential for glioma cell migration and invasion. CAL-
101, a PI3K p110δ-specific inhibitor, has been shown to
moderately reduce GBM cell proliferation and migration
without significantly affecting tumor growth in GBM
xenograft mouse models. In our experiment, another PI3K
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p110δ-specific inhibitor, PIK-294, did not significantly
influence the migration of U251 cells at concentrations
Figure 5. TG100-115 inhibited U251 cell invasion. (A) The of 60 and 120 µM (Figure S4B), likely due to the lower
representative images are from transwell experiments to detect in vitro doses and different cell lines used. In contrast, TG100-
cell invasion. Scale bar: 50 µm, magnification: ×10. (B) The invasion of 115 significantly reduced migration at the same doses
TG100-115-treated cells (150 µM) was significantly decreased compared
with DMSO (**p<0.01, Student’s t-test, n = 6). (Figure 2). This suggests that the anti-migration effect of
Abbreviations: DMSO: Dimethyl sulfoxide. TG100-115 on U251 cells may be mediated through its

A B C D

Figure 6. The underlying molecular mechanisms are mediated by TG100-115 in U251 cells. U251 cells were treated with TG100-115 (150 µm) for
24 h, and the protein expression was detected by western blotting. (A) Representative images of western blotting results. (B) TG100-115 significantly
reduced the ratio of p-Akt/t-Akt. (C) TG100-115 did not change the ratio of p-ERK/t-ERK (p>0.05). (D) TG100-115 significantly increased the ratio of
p-cofilin/t-cofilin.
Note: Statistical significance determined at **p<0.01 and ***p<0.001 using Student’s t-test, n = 4.
Abbreviations: Akt: Protein kinase B; DMSO: Dimethyl sulfoxide; ERK: Extracellular signal-regulated kinase; GAPDH: Glyceraldehyde-3-phosphate
dehydrogenase; p: Phosphorylated; t: Total.

Volume 4 Issue 3 (2025) 95 doi: 10.36922/AN025110023