Page 53 - 南京医科大学自然版

第44卷第12期 尹建星，成张淳，王协锋，等. IBSP激活NF⁃κB通路促胶质母细胞瘤恶性进展［J］.
2024年12月 南京医科大学学报（自然科学版），2024，44（12）：1662-1670 ·1667 ·

A 10 B IBSP low 100
** （%） 100 TMZ（n=13） （%） IBSP high
-
Relative IBSP expression 6 4 2 Overall survival 50 Log⁃rank P < 0.001 Overall survival 50 Log⁃rank P=0.087
TMZ（n=13）
-
8
TMZ（n=99）
+
TMZ（n=99）
+
40
100
60
-2 0 0 50 Time（months） 150 0 20 Time（months） 80 100
Pri Rec
C D E U251S N3S F U251SN3rd N3T3rd
U251S U251T3rd N3S N3T3rd Vector IBSP Vector IBSP sh⁃ctrl sh⁃IBSP sh⁃ctrl sh⁃IBSP
IBSP 40 kDa IBSP 40 kDa IBSP 40 kDa
GAPDH ** ** 35 kDa 6 ** S T3rd GAPDH ** ** 35 kDa GAPDH ** 35 kDa
5
S
1.5
5
Vector
sh⁃ctrl
Relative IBSP expression 4 3 2 Relative IBSP expression 4 2 ** Relative IBSP expression 4 3 2 Relative IBSP expression 1.0 **
IBSP
sh⁃IBSP
T3rd
0.5
0 1 0 1 0 0
U251 N3 U251 N3 U251 N3 U251SN3rd N3T3rd
G U251 N3S Vector
IBSP
Vector IBSP Vector IBSP 25 **
4 4 4 4 **
10 Q1 Q2 10 Q1 Q2 10 Q1 Q2 10 Q1 Q2
2.21 8.01 1.18 1.83 2.94 7.19 0.87 1.65 （%） 20
10 3 10 3 10 3 10 3
FL3·H∷PI 10 2 FL3·H∷PI 10 2 FL3·H∷PI 10 2 FL3·H∷PI 10 2 Apoptosis rate 15
10
1
10
Q4 Q3 10 1 Q4 Q3 10 1 Q4 Q3 10 1 Q4 Q3 5
0 78.6 11.1 0 88.1 8.90 0 78.9 10.9 0 89.2 8.32
10 10 10 10
10 0 10 1 10 2 10 3 10 4 10 0 10 1 10 2 10 3 10 4 10 0 10 1 10 2 10 3 10 4 10 0 10 1 10 2 10 3 10 4 0
FL1·H∷Annexin V FITC FL1·H∷Annexin V FITC FL1·H∷Annexin V FITC FL1·H∷Annexin V FITC U251S N3S
H U251T3rd N3T3rd sh⁃ctrl
sh⁃ctrl sh⁃IBSP sh⁃ctrl sh⁃IBSP sh⁃IBSP
4 4 4 4 25
10 Q1 Q2 10 Q1 Q2 10 Q1 Q2 10 Q1 Q2 ** **
0.91 1.70 10 3 2.84 7.08 10 3 0.97 1.44 10 3 2.57 7.46 （%） 20
3
10
FL3·H∷PI 10 2 FL3·H∷PI 10 2 FL3·H∷PI 10 2 FL3·H∷PI 10 2 15
10 1 10 1 10 1 10 1 Apoptosis rate 10
Q4 Q3 Q4 Q3 Q4 Q3 Q4 Q3
0 90.2 7.20 0 79.4 10.7 0 89.4 8.23 0 78.1 11.8 5
10 10 10 10
10 0 10 1 10 2 10 3 10 4 10 0 10 1 10 2 10 3 10 4 10 0 10 1 10 2 10 3 10 4 10 0 10 1 10 2 10 3 10 4
0
FL1·H∷Annexin V FITC FL1·H∷Annexin V FITC FL1·H∷Annexin V FITC FL1·H∷Annexin V FITC
U251T3rd N3T3rd
I Vector IBSP Vector IBSP J sh⁃ctrl sh⁃IBSP sh⁃ctrl sh⁃IBSP
TMZ 0 6 12 24 0 6 12 24 0 6 12 24 0 6 12 24 TMZ 0 6 12 24 0 6 12 24 0 6 12 24 0 6 12 24
γ⁃H2AX 15 kDa γ⁃H2AX 15 kDa
GAPDH 35 kDa GAPDH 35 kDa
U251S N3S U251T3rd N3T3rd
A：IBSP expression in primary（Pri，n=649）and recurrent（Rec n=333）glioma samples was analyzed using CGGA dataset. B：GBM samples were
divided into two groups（IBSP low and high group），prognosis of patients treated with or without TMZ was analyzed. C：IBSP expression in paired TMZ
sensitive and resistant cells was measured by Western blot. D：IBSP expression in paired TMZ sensitive and resistant cells was measured by qRT⁃PCR.
E：The validation of IBSP overexpression efficiency by Western blot. F：The validation of IBSP knockdown efficiency by Western blot. G：Control or
IBSP overexpressed sensitive cells were treated with TMZ（200 μmol/L）for 24 h，apoptotic rate was measured by flow cytometry. H：Control or IBSP
depleted resistant cells were treated with TMZ（200 μmol/L）for 24 h，apoptotic rate was measured by flow cytometry. I：Control or IBSP overexpressed
sensitive cells were treated with TMZ（200 μmol/L）for indicated time，γ⁃H2AX levels were measured by Western blot. J：Control or IBSP depleted resistant
**
cells were treated with TMZ（200 μmol/L）for indicated time，γ⁃H2AX levels were measured by Western blot. P < 0.01（n=3）.
图4 IBSP促胶质瘤替莫唑胺化疗耐药
Figure 4 IBSP promoted TMZ resistance of glioma cells