Page 23 - 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 ·1553 ·

A B C
0.5 *** 1 600 shNC
shNC 1 400 shKIF11⁃1
（g） 0.4 （min） 1 200
Tumor weight 0.2 Tumor volume 800 ***
shKIF11⁃1 0.3 1 000
600
400
0.1
0 200 0
shNC shKIF11⁃1 0 3 6 9 12 15 18 21
Time（d）
A：Tumor tissues extracted from nude mice in the shNC and shKIF11⁃1 groups after subcutaneous growth. B：Tumor weight of each group. C：Tumor
volume of each group. *** P < 0.001（n=5）.
图4 KIF11在体内促进CRC进展
Figure 4 KIF11 promoted CRC progression in vivo

ed that the IGF2BPs family is a crucial“reader”of inated the inhibitory effect on migration caused by
m6A modification，which affects gene expression by METTL3 knockdown in CRC cells（Figure 7G，H）.
［24］
regulating the stability of target mRNAs . Therefore， Consequently，KIF11 was discovered to be a down⁃
we used the starBase database to predict the relation⁃ stream target of METTL3 to facilitate CRC progression.
ship between KIF11 and their expression，and found 2.6 KIF11 promoted CRC progression through the
that the correlation between IGF2BP2 and KIF11 was PI3K/AKT signaling pathway
the highest（Supplementary Figure 2）. The RIP⁃qPCR To further explore the possible downstream molec⁃
experiment results demonstrated that IGF2BP2 and ular mechanisms of KIF11，we performed transcrip⁃
KIF11 interacted with each other（Figure 6D）. Accord⁃ tome sequencing analysis. We found a total of 166 dif⁃
ing to qRT ⁃ PCR and WB assay results，IGF2BP2 ferentially expressed genes（DEGs）between the knock⁃
knockdown decreased the mRNA and protein levels of down and control groups，comprising 114 upregulated
KIF11（Figure 6E-H）. In addition，the RNA stability genes and 52 downregulated genes in KIF11 knock⁃
assay demonstrated that KIF11 mRNA stability de⁃ down cells（P < 0.05，|log2 （fold change）| > 1）（Figure
creased after IGF2BP2 knockdown（Figure 6I，J）. 8A，B）. By gene set enrichment analysis（GSEA），we
These findings indicated that IGF2BP2 improved found that KIF11 and PI3K/AKT signaling pathway
mRNA stability and functions as a“reader”of KIF11 were closely related（Figure 8C）. WB results indicated
mRNA methylation. Rescue experiments revealed that that the knockdown of KIF11 led to significantly lower
METTL3 overexpression partially counteracted the re⁃ phosphorylation levels of PI3K and AKT than that of
duction of KIF11 expression levels caused by the NC group，whereas the total protein levels of PI3K
IGF2BP2 silencing（Figure 6K）. Taken together，MET⁃ and AKT were unaffected（Figure 8D）. In addition，
TL3 ⁃ mediated m6A modification enhances KIF11 we assessed the impact of the METTL3⁃KIF11 axis on
mRNA stability in an IGF2BP2⁃dependent manner. the PI3K/AKT pathway. We overexpressed KIF11 in
2.5 METTL3 promoted CRC proliferation and migra⁃ METTL3 ⁃ silenced colorectal cancer cells and found
tion by regulating KIF11 that upregulation of KIF11 partially recovered the
Based on these results，we further investigated the phosphorylation level of the PI3K/AKT signaling
impact and regulatory mechanisms of METTL3 on the pathway（Figure 8E）.
proliferation and migration of CRC cells. The results of Among the top 10 down ⁃ regulated downstream
CCK⁃8（Figure 7A，B），colony formation（Figure 7C，D）， genes，PROM1 attracted our attention. Previously，

and EdU（Figure 7E，F）experiments showed that MET⁃ Wei et al ［ 25 ］ reported that CD133⁃p85 interaction acti⁃
TL3 silencing inhibited the proliferative ability of CRC vated the PI3K/Akt pathway to promote tumorigenicity
cells，while overexpression of KIF11 partially rescued of glioma stem cells；Zhu et al ［26］ found that CD133 pro⁃
the adverse effect. In addition，Transwell assay results moted the chemoresistance to 5 ⁃ fluorouracil in GC
demonstrated that KIF11 overexpression partially elim⁃ cells by activating the PI3K/Akt/p70S6K pathway. So