Page 25 - 南京医科大学自然版

第45卷第11期 林书慧，钱萌森，朱 静，等. METTL3介导的KIF11 mRNA m6A修饰通过PI3K/AKT信号通路促进结
2025年11月 直肠癌进展［J］. 南京医科大学学报（自然科学版），2025，45（11）：1546-1562 ·1555 ·

A B C D
shNC IgG
1.5 shMETTL3 ** 1.5 shNC HCT116 1.5 shNC DLD1 10 ***
IGF2BP2
KIF11 enrichment 1.0 Relative KIF11 mRNA expression 1.0 shMETTL3 Relative KIF11 mRNA expression 1.0 shMETTL3 Relative folds of KIF11 8 6 4 ***
***
0.5

0 0.5 * 0.5 * 2
HCT116 DLD1 0 0 3 6 0 0 3 6 0 HCT116 DLD1

H
E shNC F Time after Actinomycin D（h） G Time after Actinomycin D（h） HCT116 DLD1
expression 1.5 shIGF2BP2 *** shNC shIGF2BP2 shNC shIGF2BP2 1.5 shIGF2BP2 shNC shIGF2BP2 shNC shIGF2BP2
DLD1
HCT116
shNC
**
1.0
1.0
Relative IGF2BP2 0.5 0 IGF2BP2 66 kDa Relative KIF11 expression 0.5 0 ** * ACTIN 130 kDa
KIF11
42 kDa
42 kDa
ACTIN
DLD1
HCT116
I 1.5 HCT116 DLD1 J 1.5 shNC DLD1 K HCT116 DLD1 shNC
Relative KIF11 mRNA expression 1.0 shIGF2BP2 *** Relative KIF11 mRNA expression 1.0 shIGF2BP2 * shNC shIGF2BP2+oe⁃NC shIGF2BP2+oe⁃METTL3 shNC shIGF2BP2+oe⁃NC shIGF2BP2+oe⁃METTL3 Relative KIF11 protein expression 1.5 ** ** ** **
shIGF2BP2+oe⁃NC
HCT116
shIGF2BP2+oe⁃METTL3
shNC
1.0
0.5
0.5
0.5
0
0
0
3
6
3
0
6
KIF11
Time after Actinomycin D（h）
Time after Actinomycin D（h）
42 kDa
ACTIN 130 kDa 0 HCT116 DLD1
A：MeRIP⁃qPCR demonstrated that METTL3 mediated m6A modification of KIF11. B，C：Effect of knockdown of METTL3 on KIF11 mRNA stabili⁃
ty. D：The binding of KIF11 to IGF2BP2 was demonstrated by RIP⁃qPCR. E，F：The knockdown efficiency of IGF2BP2 was verified by qRT⁃PCR（E）
and WB（F）. G，H：The expression level of KIF11 after the knockdown of IGF2BP2 was detected by qRT⁃PCR（G）and WB（H）. I，J：Effect of knock⁃
down of IGF2BP2 on KIF11 mRNA stability. K：The expression level of KIF11 after knockdown of IGF2BP2 and overexpression of METTL3 was detected
*
**
by WB. P < 0.05，P < 0.01，and *** P < 0.001（n=3）.
图6 METTL3介导的m6A修饰通过IGF2BP2依赖性机制增强KIF11的稳定性
Figure 6 METTL3⁃regulated m6A modification enhanced KIF11 stability through IGF2BP2⁃dependent mechanisms
EdU assays showed that KIF11 knockdown had reduced sistance development，the 5⁃year survival rate of CRC
cell proliferation ability，which could be rescued by is less than 40% ［27］ . The increasing morbidity and mor⁃
PROM1 overexpression（Figure 9E-J）. Transwell assay tality suggest that the exploration of novel biomarkers
demonstrated that overexpression of PROM1 partially is necessary for the diagnosis and prognosis of colorec⁃
rescued the decreased cell migration ability caused by tal cancer. KIF11 is a member of the kinesin family of
KIF11 knockdown（Figure 10A，B）. The WB results proteins，which plays a vital role in the mitotic pro⁃
［28- 29］
revealed that silencing of KIF11 suppressed the activa⁃ cess . Kinesin family proteins（KIFs）are a group
tion of the PI3K/AKT pathway，and the overexpression of conserved microtubule ⁃ dependent molecular motor
of PROM1 rescued the KIF11 knockdown⁃mediated in⁃ proteins that possess ATPase activity and motility prop⁃
hibition（Figure 10C）. These results suggest that erties and are mainly involved in a series of intracellular
KIF11 promotes CRC progression through the PROM1/ activities，including cytokinesis，intracellular vesicle
PI3K/AKT pathway. and organelle trafficking，and microtubule cytoskeleton
reorganization ［30- 31］ . Studies have shown that the kine⁃
3 Discussion
sin family is essential to cancer development. For ex⁃
Due to tumor recurrence，metastasis and drug re⁃ ample，KIF20A regulates malignant proliferation and