Page 17 - 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 ·1547 ·

Colorectal cancer（CRC）is the third most com⁃ In this study，we demonstrated that KIF11 was
mon cancer globally and the second leading cause of highly expressed in CRC，and its knockdown inhibited
［1］
death from cancer . According to GLOBOCAN 2020 the proliferation and migration of CRC cells. We also
estimates，3 154 674 new CRC cases are projected found that the METTL3 ⁃ IGF2BP2 axis regulated
［2］
worldwide by 2040 ，highlighling its significance as a KIF11 in an m6A⁃dependent way. In addition，KIF11
public health crisis. Despite advances in diagnostic promoted CRC progression by regulating PROM1 and
and therapeutic strategies，patient prognosis remains PI3K/AKT signaling pathway. Collectively，these find⁃
［3］
poor . Thus，exploring the molecular mechanisms of ings position KIF11 as a potential therapeutic target
CRC and promising therapeutic targets are essential. and prognostic biomarker for CRC.
Kinesin family member 11（KIF11），a plus⁃end⁃
1 Materials and Methods
directed kinesin motor protein of the kinesin family，
plays a critical role in spindle pole separation and chro⁃ 1.1 Materials
［4］
mosome alignment during mitosis . Accumulating evi⁃ 1.1.1 Clinical samples
dence denonstrates that KIF11 upregulation is associat⁃ This study collected 30 pairs of CRC tissues and
ed with poor prognosis in many cancers. For example， adjacent normal tissues from patients undergoing sur⁃

KIF11 is highly expressed and promotes cell prolifera⁃ gery at the Second Affiliated Hospital of Nanjing Medi⁃
［5］
tion in gallbladder cancer ；in hepatocellular carcino⁃ cal University. Before surgery，none of the patients
ma，KIF11 is negatively correlated with senescence underwent radiotherapy or chemotherapy. All patients
biomarkers；its knockdown induces cellular senes⁃ who participated in this study signed informed consent
cence and suppresses hepatocellular carcinoma（HCC） forms. The whole process was in strict accordance with
［6］
progression . Epigenetic modifications，particularly the Declaration of Helsinki. The ethical approval of
N6 ⁃ methyladenosine（m6A），are pivotal in disease this study was provided by the Institutional Review
pathogenesis. As the most prevalent internal mRNA Board of Nanjing Medical University（Ethics No. 2024⁃
modification in humans（ affecting almost 90% of KY⁃274⁃01）.
［7］
transcripts） ，m6A regulates post⁃transcriptional pro⁃ 1.1.2 Animals
cesses. However，whether m6A modifications and their Four ⁃ week ⁃ old，16-18 g specific pathogen free
regulatory enzymes control KIF11 expression in CRC （SPF）male BALB/c nude mice were purchased from
remains unexplored. GemPharmatech Co.，Ltd. The nude mice were housed
The dynamic regulation of m6A methylation modi⁃ in an environment of 20-27 ℃ with relative humidity
fication is performed by methyltransferases（“writers”）， of 40%-60%. The Nanjing Medical University Animal
demethylases（“erasers”），and binding proteins（“read⁃ Ethics Committee granted its approval for animal stud⁃
［8］
ers”） . METTL3 is one of the major methyltransferases ies（IACUC⁃2410071）.
proven to be highly expressed in various cancers and to 1.1.3 Reagents and instruments
accelerate tumor malignant development，including TRIzol（Invitrogen，USA）；anti ⁃ KIF11，anti ⁃
［9-12］
colorectal and lung cancers . Similarly，IGF2BP2 is IGF2BP2，anti ⁃ ACTIN，anti ⁃ GAPDH（Proteintech，
a key m6A modification“reader”that encourages the USA）；anti⁃METTL3（Youmengbiology，China）；anti⁃
growth of malignancies by affecting the stability and PI3K，anti⁃p⁃PI3K，anti⁃AKT（CST，USA）；p⁃AKT（Af⁃
［13-15］
translation of downstream target mRNAs . finity，USA）；anti⁃IgG（Millipore，USA）；SYBR Green
TM
PROM1，also known as CD133，is a cancer stem premix（CWBIO，Taizhou，China）；BeyoClick EdU
cell（CSC）marker involved in tumorigenesis，drug re⁃ cell proliferation kit with AF555（Beyotime，Shanghai，
sistance，and apoptosis in CSCs. For example，CD133 China）；Annexin V⁃FITC/PI Apoptosis Detection Kit
［16］
can promote angiogenesis through VEGF⁃A ；it can （Vazyme，Nanjing，China）；Magna RIP kit，anti⁃m6A，
also promote gastric cancer（GC）progression through IgG antibody（Millipore，USA）. Light Cycler480Ⅱreal⁃
［17］
the PI3K/AKT signaling pathway . time fluorescence PCR Instrument（Roche，USA）；Cen⁃