Page 71 - 《中国药科大学学报》2026年第1期

第 57 卷第 1 期 王俪颖，等：Isthmin-1 通过调控 FoxO 信号通路抑制非小细胞肺癌细胞生长的研究 65
A KEGG enrichment ScatterPlot B GO_term: molecular function

hsa04110 | Cell cycle GO:0005515 | protein binding
hsa05168 | Herpes simplex virus 1 infection GO:0046872 | metal ion binding
Gene number Gene number
hsa03013 | Nucleocytoplasmic transport 50 GO:0008017 | microtubule binding 200
400
100
hsa04140 | Autophagy-animal 150 GO:0000981 | DNA-binding transcription factor activity 600
hsa05210 | Colorectal cancer 200 GO:0016740 | transferase activity 800
hsa05220 | Chronic myeloid leukemia P value GO:0000978 | RNA polymerase Il cis-regulatory region P value
hsa05215 | Prostate cancer 1e−06 GO:0000976 | transcription cis-regulatory region binding 2.5e−08
hsa04115 | p53 signaling pathway 2e−06 GO:0000166 | nucleotide binding 5.0e−08
7.5e−08
hsa04120 | Ubiquitin mediated proteolysis GO:0003700 | DNA-binding transcription factor activity
hsa04068 | FoxOsignaling pathway GO:1990837 | sequence-specific double-stranded DNA binding
0.4 0.5 0.050 0.075 0.100 0.125 0.150
Rich factor Rich factor
C GO_term: biological process D GO_term: cellular component

GO:0006695 | cholesterol biosynthetic process GO:0005737 | cytoplasm
GO:0016126 | sterol biosynthetic process Gene number GO:0005829 | cytosol Gene number
20
GO:0000122 | negative regulation of transcription by RNA polymerase II 40 GO:0005856 | cytoskeleton 100
60 200
GO:0007399 | nervous system development GO:0005634 | nucleus 300
80
GO:0045944 | positive regulation of transcription by RNA polymerase II GO:0000785 | chromatin 1 400
100
GO:0007049 | cell cycle GO:0030139 | endocytic vesicle P value
P value
GO:0008203 | cholesterol metabolic process GO:0005886 | plasma membrane 2e−06
2e−09
GO:0006694 | steroid biosynthetic process 4e−09 GO:0042995 | cell projection 4e−06
GO:0051301 | cell division 6e−09 GO:0070161 | anchoring junction 6e−06
8e−09
GO:0008202 | steroid metabolic process GO:0005794 | Golgi apparatus
0.1 0.2 0.3 0.4 0.5 0.05 0.10 0.15 0.20
Rich factor Rich factor
Figure 4 rISM1 regulated FoxO signaling pathway
H1299 cells were treated with 1.0 μg/mL rISM1 for 24 h, and subjected to RNA sequencing. David website was used for KEGG (A), GO-MF (B),
GO-BP (C), GO-CC (D) analyasis. The first 10 enriched terms were shown for each anlyasis
A B A549 H1299

250 *** 2.0 ** ** 2.5 ** ***
Control Control Control
200 rISM1 1.5 rISM1 2.0 rISM1
FPKM 150 ** * Relative mRNA expression 1.0 Relative mRNA expression 1.5
10
1.0
5 * * 0.5
* 0.5
0 0 FoxO3 FoxO1 0 FoxO3 FoxO1
CDKN1A CDKN1B PIK3CD FoxO3 FoxO1 PTEN

C A549 H1299 D 1.5 ** ** E 4
Relative ROS production Relative caspase-3 activity 2
rISM1
rISM1
rISM1 − + − + Control 3 *** *** Control
FoxO3 90 kD 1.0
FoxO1 74 kD 0.5 1

GAPDH 36 kD 0 0
A549 H1299 A549 H1299
Figure 5 rISM1 upregulated FoxO1 and FoxO3 expression, increased ROS production, and induced cell apoptosis ( x ± s，n=3)
A: Gene expression levels (FRPK) according to RNA sequencing data; B-E: Cells were treated with 1.0 μg/mL rISM1 for 24 h, and subjected to
qRT-PCR assay (B) Western blot assay (treatment for 48 h) (C), ROS assay (D), and caspase-3 activity assay (E)
* P<0.05, **P<0.01, ***P<0.001
表明，FoxO 作为肿瘤细胞存活的关键分子 Akt 的 挥重要作用 [16−17] 。进一步检测细胞内 ROS 的产生
下游信号分子，在线粒体损伤诱导的细胞凋亡中发 和 Caspase-3 的活性，结果显示 rISM1 处理组 ROS