南方医科大学学报 ›› 2026, Vol. 46 ›› Issue (7): 1533-1542.doi: 10.12122/j.issn.1673-4254.2026.07.07
• • 上一篇
王琳钰1,2(
), 王晓燕1,2,4, 朱季军1,2, 蒋波1,2, 徐洁1,2,4, 高彩月2,3,4, 李冬冬2,3,4, 王玉2,3,4, 李晓敏1,2,4(
)
收稿日期:2026-01-26
出版日期:2026-07-20
发布日期:2026-07-20
通讯作者:
李晓敏
E-mail:1007080916@qq.com;lxm1980326@sina.com
作者简介:王琳钰,在读硕士研究生,E-mail: 1007080916@qq.com
基金资助:
Linyu WANG1,2(
), Xiaoyan WANG1,2,4, Jijun ZHU1,2, Bo JIANG1,2, Jie XU1,2,4, Caiyue GAO2,3,4, Dongdong LI2,3,4, Yu WANG2,3,4, Xiaomin LI1,2,4(
)
Received:2026-01-26
Online:2026-07-20
Published:2026-07-20
Contact:
Xiaomin LI
E-mail:1007080916@qq.com;lxm1980326@sina.com
Supported by:摘要:
目的 探究丝切蛋白1(CFL1)及其磷酸化形式p-CFL1在结肠癌中的表达情况,分析其与临床病理特征及预后的关系。揭示CFL1/p-CFL1调控结肠癌细胞迁移的分子机制,重点阐明其核定位在此过程中所发挥的作用及潜在机制。 方法 利用TCGA数据库、结肠癌组织芯片免疫组化分析CFL1和p-CFL1表达及临床病理参数相关性;应用Kaplan-Meier生存分析探究CFL1和p-CFL1表达水平与结肠癌患者生存期的关系;应用免疫荧光明确CFL1和p-CFL1在细胞核和细胞质的定位;构建野生型(CFL1-WT)与核定位信号突变型(CFL1-MUT)载体,在HCT116细胞中过表达,通过Transwell小室迁移实验检测细胞迁移能力;结合转录组测序与KEGG富集分析解析差异基因与信号通路。 结果 免疫组化结果显示,CFL1与p-CFL1在结肠癌组织中表达显著高于癌旁组织(P<0.0001),且二者表达呈正相关(r=0.4753,P<0.0001)。CFL1和p-CFL1高表达与淋巴结转移、临床分期晚及患者总生存期缩短相关(P<0.05)。免疫组化与免疫荧光显示CFL1/p-CFL1可定位于细胞质与细胞核。功能实验表明,过表达CFL1-WT与CFL1-MUT均能促进HCT116细胞迁移,且CFL1-WT作用更强。KEGG分析提示,CFL1-WT调控的基因显著富集于Ras、PI3K-Akt、细胞粘附分子等肿瘤转移相关通路。 结论 CFL1/p-CFL1在结肠癌中高表达,是预后不良的潜在生物标志物。CFL1可通过核定位进一步激活肿瘤转移相关信号通路,从而促进结肠癌迁移。
王琳钰, 王晓燕, 朱季军, 蒋波, 徐洁, 高彩月, 李冬冬, 王玉, 李晓敏. CFL1及p-CFL在结肠癌中高表达预示结肠癌患者预后不良并促进癌细胞迁移[J]. 南方医科大学学报, 2026, 46(7): 1533-1542.
Linyu WANG, Xiaoyan WANG, Jijun ZHU, Bo JIANG, Jie XU, Caiyue GAO, Dongdong LI, Yu WANG, Xiaomin LI. High expression of CFL1 and phospho-CFL1 predicts poor prognosis and promotes migration of colon cancer cells in vitro[J]. Journal of Southern Medical University, 2026, 46(7): 1533-1542.
图1 结肠癌及癌旁组织CFL1的表达情况
Fig.1 CFL1 is highly expressed in colon cancer tissues. A: Expression levels of CFL1 in colon adenocarcinoma (COAD) and rectal adenocarcinoma (READ) from the TCGA database. B, C: Representative immunohistochemical staining and the corresponding H-score analysis of CFL1 in colon cancer tissues and paired adjacent normal tissues (n=80). Scale bar=500 µm (upper panel); Scale bar=200 µm (lower panel). *P<0.05, ****P<0.0001.
图2 结肠癌及癌旁组织p-CFL1的表达情况
Fig.2 p-CFL1 is highly expressed in colon cancer tissues. A, B: Representative immunohistochemical staining and the corresponding H-score analysis of p-CFL1 in colon cancer tissues and paired adjacent normal tissues (n=80). Scale bar=500 µm (upper panel); Scale bar=200 µm (lower panel). ****P<0.0001.
图3 CFL1和p-CFL1在结肠癌组织中的表达呈正相关
Fig.3 Positive correlation between expressions of CFL1 and p-CFL1 in colon cancer tissues. A: Representative images showing consistent expression trends of CFL1 and p-CFL1 in the same colon cancer tissue samples (Scale bar=100 μm). B: Pearson correlation analysis between CFL1 and p-CFL1 expression levels in colon cancer tissues (n=96).
图4 CFL1和p-CFL1高表达对结肠癌患者术后生存情况的影响
Fig.4 Impact of high expression of CFL1 and p-CFL1 on overall survival of patients with colon cancer. A: Kaplan-Meier survival curves comparing overall survival between patients with high and low expression of CFL1 (P=0.0158). B: Kaplan-Meier survival curves comparing overall survival between patients with high and low expression of p-CFL1 (P=0.0030).
图5 CFL1和p-CFL1在部分结肠癌的细胞核中有阳性表达
Fig.5 Nuclear localization of CFL1 and p-CFL1 in a subset of colon cancer cases. A: Representative immunohistochemical images showing nuclear and cytoplasmic staining of CFL1 and p-CFL1 in colon cancer tissues (Scale bar=100 μm). B: Immunofluorescence analysis confirms the localization of CFL1 and p-CFL1 in both the nucleus and cytoplasm of HCT116 cells (Scale bar=25 μm).
图6 过表达野生型和NLS突变型CFL1均促进结肠癌细胞迁移
Fig.6 Overexpression of wild-type and NLS-mutant CFL1 promotes migration of colon cancer cells. A: Sanger sequencing verification showing the presence of the NLS in the CFL1-WT vector and its mutation in the CFL1-MUT vector. B: Western blotting confirms overexpression of CFL1 in HCT116 cells transfected with CFL1-WT and CFL1-MUT vectors. C: Immunofluorescence staining showing that wild-type CFL1 overexpression leads to both nuclear and cytoplasmic distribution, while the NLS-mutant confines CFL1 to the cytoplasm (Scale bar=20 μm). D: Transwell migration assays demonstrating that both CFL1-WT and CFL1-MUT enhancemigration of colon cancer cells (Scale bar=100 μm; n=5). *P<0.05, **P<0.01.
图7 野生型和NLS突变型CFL1的KEGG富集分析
Fig.7 KEGG enrichment analysis of wild-type and NLS-mutant CFL1. A, B: Volcano plots displaying the numbers of differentially expressed genes (DEGs) identified by RNA-seq in HCT116 cells transfected with CFL1-MUT, CFL1-WT, or control vectors. C, D: Bubble plots of KEGG pathway enrichment analysis for up-regulated DEGs (MUT vs NC: 63, WT vs NC: 41) in the CFL1-MUT and CFL1-WT groups, respectively. E, F: Bubble plots of KEGG pathway enrichment analysis for down-regulated DEGs (MUT vs NC: 41, WT vs NC: 54) in the CFL1-MUT and CFL1-WT groups, respectively.
图8 核内定位的CFL1的KEGG富集分析
Fig.8 KEGG enrichment analysis of nuclear-localized CFL1. A, B: Venn diagram and heatmaps showing the 31 up-regulated and 45 down-regulated differentially expressed genes in CFL1-WT group compared to the control. C, D: KEGG enrichment analysis of the 31 up-regulated and 45 down-regulated DEGs.
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