Page 55 - 南京医科大学自然版

第45卷第8期 刁庆飞，张 昊，杨春白雪，等. EZH2靶向FAK/F⁃actin/ROS信号通路影响结直肠癌进展的机制研究［J］.
2025年8月 南京医科大学学报（自然科学版），2025，45（8）：1110-1122 ·1111 ·

［Abstract］ Objective：To explore the regulatory effect of the enhancer of Zeste homolog 2（EZH2）on the focal adhesion kinase
（FAK）/filamentous actin（F⁃actin）/reactive oxygen species（ROS）pathway，and to analyze its effect on the proliferation，invasion，and
metastasis of colorectal cancer（CRC）cells. Methods：Tissue samples from 50 patients undergoing CRC resection at the First Affiliated
Hospital of Hebei North University were collected，including both cancer tissues and adjacent normal tissues. Immunohistochemistry
was used to detect EZH2 expression，and clinical data were analyzed to determine the correlation between EZH2 expression and
clinicopathological parameters as well as survival prognosis. A subcutaneous xenograft model using CRC nude mice was established，

dividing the mice into the negative control（EZH2 NC）group，EZH2 overexpression（EZH2 mimic）group，EZH2 NC+cytochalasin D
group，and EZH2 mimic+cytochalasin D group. After 14 days of cultivation，the tumor growth was observed. Human CRC cell lines
SW480 and SW620 cells were cultured in vitro and divided into the same four groups using lipofection. Western blot was used to detect
the expression of FAK，F⁃actin，and ROS pathway⁃related proteins. Immunofluorescence staining was used to observe F⁃actin expression
and distribution. Wound healing，transwell，and CCK⁃8 assays were used to assess cell migration，invasion，and viability. ChIP⁃qPCR
was used to detect the enrichment of FAK，NADPH oxidase（NOX）⁃2，and NOX4 on EZH2. Results：Immunohistochemistry analysis
showed the expression levels of EZH2 were significantly higher in CRC tissues than in adjacent normal tissues（P < 0.05）. EZH2
expression levels were closely associated with lymph node metastasis and distant transfer events（all P < 0.05）. The analysis of follow⁃up
data showed that the 5 ⁃ year overall survival rate of CRC patients with low EZH2 expression was significantly higher than that of
patients with high EZH2 expression，with a statistically significant difference（P < 0.05）. The subcutaneous CRC xenograft model was
successfully established，with the EZH2 mimic group showing significantly larger tumor volumes than that of the EZH2 NC group（P <
0.05）. Post cytochalasin D intervention，both EZH2 NC+cytochalasin D and EZH2 mimic+cytochalasin D groups showed significantly
reduced tumor volumes compared with the untreated groups（P < 0.05），though no significant difference was noted between the two
intervention groups（P > 0.05）. The expression levels of EZH2，p⁃FAK，p⁃Paxillin，NOX2，NOX4，and p⁃JNK proteins in the EZH2
mimic group were significantly higher than those in the EZH2 NC group（P < 0.05），while the expression levels of RUNX family
transcription factor 3（RUNX3）protein were slightly lower than that in the EZH2 NC group（P < 0.05）. The number of F ⁃ actin
distribution，migration ability，and cell viability increased in the EZH2 mimic group than in the EZH2 NC group. After the intervention
of cytochalasin D，there was no significant difference in the expression levels of EZH2，p⁃FAK，and p⁃Paxillin proteins compared with
the untreated group（P > 0.05），while the expression levels of NOX2，NOX4，and p⁃JNK proteins were significantly decreased（P <
0.05），and the expression levels of RUNX3 protein were significantly increased compared with the untreated group（P < 0.05）. There
was no significant difference between the two intervention groups（P > 0.05）. In addition，the number of F⁃actin distribution，migration
ability，and cell viability were significantly decreased in the intervention groups，with no significant difference between the two groups
（P > 0.05）. The results of the ChIP⁃qPCR assay showed that after using the EZH2 antibody，the promoter contents enriched by FAK，
NOX2，and NOX4 were significantly increased，respectively（P < 0.05）. Conclusion：EZH2 promotes the proliferation，invasion，and
metastasis of CRC cells by upregulating the activity of the FAK/F⁃actin/ROS pathway.
［Key words］ EZH2；FAK；F⁃actin；ROS；colorectal cancer；proliferation；invasion；migration
［J Nanjing Med Univ，2025，45（08）：1110⁃1122］

结直肠癌（colorectal cancer，CRC）是一种常见 物 2（polycomb repressive complex 2，PRC2）的核心成
［1］
的严重危害人类生命的消化系统恶性肿瘤 。CRC 分。越来越多的证据表明EZH2与多种癌症相关，包
在女性和男性中发病率分别位居第 2 位和第 3 位。 括鼻咽癌、乳腺癌、肺癌和膀胱癌 ［6-8］ 。此外，EZH2
尽管诊断和治疗方法有所改进提升，但40%~50%的 过表达提示肿瘤更具侵袭性，与预后不良有关 ［9-10］ 。
CRC 患者会出现肿瘤的复发转移进而导致预后不 近年来，多项研究表明 EZH2 在 CRC 中的表达
［2］
良 。因此，明确CRC发生和转移的分子机制，开发 水平与肿瘤的进展密切相关。EZH2 的过表达与
有效的 CRC 生物标志物和改进治疗策略是目前临 CRC 的细胞增殖能力增强、侵袭转移倾向增加、化
床研究的重点。 疗耐药性以及不良预后有关。EZH2在CRC中的作

Zeste同源物增强子2（enhancer of Zeste homolog 用机制被认为是通过沉默多个抑癌基因如钙黏蛋
［3］ ［4］
2，EZH2）在细胞周期进程 、自噬和凋亡 以及DNA 白E（E⁃cadherin，E⁃Cad）、周期蛋白依赖的激酶抑制
损伤修复 中发挥重要作用，EZH2是多梳抑制复合 剂（p16INK4a，p16）等，促进肿瘤细胞的表型转化，
［5］