Page 44 - 南京医科大学自然版

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·782 · 南 京 医 科 大 学 学 报 2025年6月

A B 1.5 D 1.5
siNC siFBXO6⁃1 siFBXO6⁃2 siFBXO6⁃3 mRNA levels protein levels
FBXO6 487 bp 1.0 1.0
GAPDH 203 bp 0.5 ** FBXO6 0.5 **

siFBXO6⁃2
C Relative FBXO6 Relative
siNC siFBXO6⁃1 siFBXO6⁃3 0 siNC 0 siNC
siFBXO6⁃2
siFBXO6⁃2
FBXO6 33 kDa siFBXO6⁃1 siFBXO6⁃3 siFBXO6⁃1 siFBXO6⁃3
β⁃actin 42 kDa
Relative cell proliferation rate 1.0 ** ×100 Relative cell area/ （%） total area 40 **
E 1.5 F siNC siFBXO6 G 60

0.5
20
0
0
siNC siFBXO6
siNC siFBXO6
U87 cells were transfected with siNC，siFBXO6⁃1，siFBXO6⁃2 or siFBXO6⁃3 for 48 h. A-D：The mRNA and protein levels of FBXO6 were
checked by RT⁃PCR（A：electrophoretic bands of RT⁃PCR；B：statistical chart）and Western blot（C：electrophoretic bands of Western blot；D：statistical
chart）；E-G：The levels of cell proliferation and invasion were detected by CCK⁃8（E）and Transwell experiment（F：representative images，scale bar=
**
200 μm；G：statistical chart）. Compared with the siNC transfection group，P < 0.01（n=3）.
图3 沉默FBXO6对U87细胞增殖和侵袭的影响
Figure 3 Effects of FBXO6 silencing on proliferation and invasion of U87 cells
A B
DMSO + - - -
U0126 - + - -
SP600125 - - + - p⁃ERK1/2/t⁃ERK1/2
Perifosine - - - + p⁃JNK/t⁃JNK
Relative protein levels 1.5
p⁃ERK1/2 44/42 kDa p⁃Akt1/t⁃Akt1
t⁃ERK1/2 44/42 kDa 1.0
p⁃JNK 54/46 kDa **
t⁃JNK 54/46 kDa 0.5 ** **
p⁃Akt1 55 kDa
0
DMSO U1206 SP600125 Perifosine
t⁃Akt1 55 kDa
β⁃actin 42 kDa
C D E F
Perifosine
Perifosine
DMSO U0126 SP600125 mRNA levels 1.5 DMSO U0126 SP600125 protein levels 1.5
FBXO6 487 bp 1.0 FBXO6 33 kDa 1.0
Relative FBXO6 Relative FBXO6
GAPDH 203 bp 0.5 ** β⁃actin 42 kDa 0.5 **

0
SP600125
DMSO U1206 Perifosine 0 DMSO U1206 Perifosine
SP600125
U87 cells were treated with DMSO，U0126，SP600125 or Perifosine. A，B：The phosphorylation levels of ERK1/2，JNK and Akt1 were examined by
Western blot（A：electrophoretic bands of Western blot；B：statistical chart）；C-F：The mRNA and protein levels of FBXO6 were detected by RT⁃PCR
（C：electrophoretic bands of RT⁃PCR；D：statistical chart）and Western blot（E：electrophoretic bands of Western blot；F：statistical chart）. Compared
**
with the DMSO treatment group，P < 0.01（n=3）.
图4 ERK1/2、JNK和Akt1抑制剂对U87细胞FBXO6表达的影响
Figure 4 The effects of ERK1/2，JNK and Akt1 inhibitors on FBXO6 expression in U87 cells