Page 82 - 《中国药科大学学报》2026年第1期

76 学报 Journal of China Pharmaceutical University 2026, 57(1): 68 − 77 第 57 卷

A B ** 150 ** C Penfluridol - + - - + - - + - - + + - + + - - - - + - - + - - + - - + + - + + -

si-NC -
A375 cell survival/% 150 ** B16 cell survival/% 150 ** A375 cell survival/% 150 B16 cell survival/% 100 si-HSPA6 -
100
100
100
HSPA6
50
50
50
50
p21
0
p53
Control OE-NC OE-HSPA6 0 Control OE-NC OE-HSPA6 0 Control si-NC si-HSPA6 0 Control si-NC si-HSPA6 β-actin
D A375 B16
A375 60
Control Penfluridol si-NC si-NC+Penfluridol si-HSPA6 si-HSPA6+Penfluridol **
10 5 0.3% 2.2% 10 5 2.1% 5.8% 10 5 0.4% 2.8% 10 5 2.2% 5.5% 10 5 0.4% 3.3% 10 5 2.1% 4.0% 40
10 4 10 4 10 4 10 4 10 4 10 4 20
PI 10 3 10 3 10 3 10 3 10 3 10 3 A375 apoptotic cells/% 0
si-HSPA6+Penfluridol
Penfluridol
0 10 2 0.6% 0 10 2 32.2% 0 10 2 0.5% 0 10 2 28.4% 0 10 2 0.5% 0 10 2 6.1% Control si-NC si-HSPA6
0 10 2 10 3 10 4 10 5 0 10 2 10 3 10 4 10 5 0 10 2 10 3 10 4 10 5 0 10 2 10 3 10 4 10 5 0 10 2 10 3 10 4 10 5 0 10 2 10 3 10 4 10 5 si-NC+Penfluridol
Annexin-V FITC
E
B16 40
Control Penfluridol si-NC si-NC+Penfluridol si-HSPA6 si-HSPA6+Penfluridol **
10 5 1.1% 3.1% 10 5 1.9% 2.2% 10 5 0.8% 3.0% 10 5 1.8% 4.1% 10 5 1.1% 2.1% 10 5 2.7% 7.0% 30
20
10 4 10 4 10 4 10 4 10 4 10 4 10
PI B16 apoptotic cells/%
10 3 10 3 10 3 10 3 10 3 10 3 0
si-NC+Penfluridol
10 2 10 2 10 2 10 2 10 2 10 2 Control si-NC
0 3.6% 0 20.9% 0 3.4% 0 20.0% 0 0.4% 0 4.3% Penfluridol si-HSPA6
0 10 2 10 3 10 4 10 5 0 10 2 10 3 10 4 10 5 0 10 2 10 3 10 4 10 5 0 10 2 10 3 10 4 10 5 0 10 2 10 3 10 4 10 5 0 10 2 10 3 10 4 10 5 si-HSPA6+Penfluridol
Annexin-V FITC
F
A375
Control Penfluridol si-NC si-NC+Penfluridol si-HSPA6 si-HSPA6+Penfluridol
Dip G1 48.62% Dip G1 62.12% Dip G1 48.37% Dip G1 60.51% Dip G1 48.10% Dip G1 52.04%
80
Dip S 38.76%
Dip S 30.55% 700 Dip S 23.59% 600 Dip S 30.75% 700 Dip S 25.00% Dip S 30.20% 400 Dip G2 9.20% Cell numbers in G1 Cycle/%100 *
Dip G2 21.70%
100 200 300 400 500 500 400 500 300 20 0
Dip G2 20.88%
Dip G2 14.49%
Dip G2 14.29%
Dip G2 20.83%
60
40
Number Number 300 Number 200 Number 300 Number 100 200 300 400 500 Number 200 Control si-NC+Penfluridol
si-NC
Penfluridol
si-HSPA6
0 100 0 0 100 0 0 100 0 si-HSPA6+Penfluridol
0 50 100 150 200 0 50 100 150 200 0 50 100 150 200 0 50 100 150 200 0 50 100 150 200 0 50 100 150 200
Propidium lodide Propidium lodide Propidium lodide Propidium lodide Propidium lodide Propidium lodide
G
B16
Control Penfluridol si-NC si-NC+Penfluridol si-HSPA6 si-HSPA6+Penfluridol
Dip G1 41.12% Dip G1 75.31% Dip G1 44.04% 700 Dip S 29.48% Dip G1 46.90% 800 Dip S 43.41% in G1 Cycle/%100 **
Dip G1 56.02%
Dip G1 70.52%
80
100 200 300 400 500 Dip S 57.32% 200 400 600 8001000 Dip S 21.62% 100200300400500600 Dip S 52.26% 500 100200300400500600 Dip S 53.10% 600 Cell numbers 40 0
Dip G2 0.57%
Dip G2 0.00%
Dip G2 3.70%
Dip G2 3.07%
Dip G2 1.56%
Dip G2 0.00%
60
20
Number Number Number Number 300 Number Number 400 Control si-NC+Penfluridol
Penfluridol
si-NC
si-HSPA6
0 0 0 100 0 0 200 0 si-HSPA6+Penfluridol
0 50 100 150 0 50 100 150 0 50 100 150 0 50 100 150 0 50 100 150 0 50 100 150
Propidium lodide Propidium lodide Propidium lodide Propidium lodide Propidium lodide Propidium lodide
Figure 6 Penfluridol-induced A375 and B16 cell apoptosis and cell cycle arrest are dependent on the presence of HSPA6( x ± s, n=3)
A: Effect of high HSPA6 expression on A375 and B16 cell survival rate；B: Effect of low HSPA6 expression on A375 and B16 cell survival rate；C:
Expression of relevant proteins in A375 and B16 cells following treatment with penfluridol and/or HSPA6-specific siRNA；D, E: Apoptosis of A375
and B16 cells under different treatments detected by flow cytometry；F, G: Cell cycle distribution of A375 and B16 cells under different treatments
detected by flow cytometry
OE-NC: over-expression negative control plasmid; OE-HSPA6: HSPA6 over-expression plasmid; si-NC: small interfering RNA lentivirus negative
control; si-HSPA6: HSPA6 gene-silencing small interfering RNA lentivirus
*P < 0.05, **P < 0.01
现象高度吻合。更重要的是，p53 的激活可通过双 另一方面通过激活 caspase 级联反应（如 cleaved
重机制诱导凋亡：一方面通过调控 Bcl-2 家族蛋白 caspase-3 的上调）执行细胞凋亡程序 。这为解释
[20]
（如：降低 Bcl-2/Bax 比值）促进线粒体凋亡途径 ； HSPA6 敲低导致凋亡率下降提供了分子基础——
[19]