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A B C
IP: IgG S100A4 IP: IgG HA IP: IgG S100A4
IB 25 kDa— SUMO1⁃S100A4 IB 15 kDa— Flag⁃UBC9 IB 25 kDa— SUMO1⁃S100A4
10 kDa— S100A4 10 kDa— HA⁃S100A4 10 kDa— S100A4
Input Input Input
10 kDa— S100A4 15 kDa— Flag⁃UBC9 10 kDa— S100A4
15 kDa— UBC9 10 kDa— HA⁃S100A4 15 kDa— Flag⁃UBC9
70 kDa— SENP1 40 kDa— β⁃actin 70 kDa— Flag⁃SENP1
70 kDa— SENP2 Flag⁃UBC9： + + 40 kDa— β⁃actin
pcDNA3.1： + -
70 kDa— SENP3 HA⁃S100A4： - + pcDNA3.1： + + - -
Flag⁃UBC9： - - + -
40 kDa— β⁃actin IP: IgG HA Flag⁃SENP1： - - - +
shScr：+ + - - - -
shUBC9：- - + - - - IB 70 kDa— Flag⁃SENP1
shSENP1：- - - + - - 10 kDa— HA⁃S100A4
shSENP2：- - - - + -
shSENP3：- - - - - + Input
70 kDa— Flag⁃SENP1
10 kDa— HA⁃S100A4
40 kDa— β⁃actin
Flag⁃SENP1： + +
pcDNA3.1： + -
HA⁃S100A4： - +
A：In MIA PaCa⁃2 cells with knockdown of UBC9，SENP1，SENP2 or SENP3 by shRNA，protein A/G magnetic beads were conjugated with anti⁃
S100A4 antibody and precipitated with cell lysates，and the degree of SUMOylation was detected by anti⁃SUMO1 antibody. B：The interaction between
S100A4 and UBC9 or SENP1 was detected by co⁃immunoprecipitation. C：S100A4 SUMOylation was detected with an anti⁃SUMO1 antibody in MIA
PaCa⁃2 cells transfected with Flag⁃UBC9 or Flag⁃SENP1.
图5 SUMO化相关酶UBC9和SENP1参与MIA PaCa⁃2细胞中S100A4 蛋白的SUMO化与去SUMO化修饰过程
Figure 5 UBC9 and SENP1 are involved in the SUMOylation and deSUMOylation of S100A4 protein in MIA PaCa⁃2 cells

A B
Nucleus Nucleus
10 kDa— S100A4 10 kDa— S100A4
40 kDa— β⁃actin 40 kDa— β⁃actin

70 kDa— Lamin B1 70 kDa— Lamin B1
Cytoplasm Cytoplasm
10 kDa— S100A4 10 kDa— S100A4
40 kDa— β⁃actin 40 kDa— β⁃actin
70 kDa— Lamin B1 70 kDa— Lamin B1
shScr shUBC9 shSENP1 WT K22R K96R DKR

A：Results of protein immunoblotting using cytoplasmic and nuclear extracts. The expression of endogenous S100A4 in the nucleus and cytoplasm
of shUBC9，shSENP1 and negative control MIA PaCa⁃2 cells was detected. B：Expression of S100A4 in the nucleus and cytoplasm of cells transfected
with different mutant forms of S100A4.
图6 S100A4在不同处理的胰腺癌细胞中胞核与胞质表达情况
Figure 6 Expression of S100A4 in the nucleus and cytoplasm of pancreatic cancer cells with different treatment

细胞迁移侵袭的能力。 蛋白质参与 ［19］ 。1989 年，Ebralidze ［20］ 、Ishikawa ［21］ 等
鉴定出S100A4，并首次证明其与肿瘤转移相关。最
3 讨 论
早在人乳腺癌中发现 S100A4 的过表达可作为预后
肿瘤转移是影响预后的关键因素，也是导致治 不良和高转移潜力的指标 ［22］ 。对于食管鳞状细胞
疗失败和患者死亡的首要原因。癌症转移是一个 癌、胃癌、结直肠癌、肝细胞癌、PC 和胆囊癌 ［23-24］ 等
多步骤的过程，在分子水平上有许多与转移相关的 消化系统肿瘤，S100A4是其不良预后和手术切除或