CYFIP1对结直肠癌细胞HT29增殖和凋亡的影响  

Effect of CYFIP1 on proliferation and apoptosis of colorectal cancer cell HT29

作　　者：余富龙 李亮 强昊 袁慧 王松[1] 程晓虎[1] 江闰犇 杨雅茹[3] 刘志宁[1] YU Fu-long;LI Liang;QIANG Hao;YUAN Hui;WANG Song;CHENG Xiao-hu;JIANG Run-ben;YANG Ya-ru;Liu Zhi-ning(Dept of General Surgery,the Second Affiliated Hospital of Anhui Medical University,Hefei 230000,China;Anhui Medical University,Hefei 230032,China;Drug Clinical Trial Research Center,the Second Affiliated Hospital of Anhui Medical University,Hefei 230000,China)

机构地区：[1]安徽医科大学第二附属医院普外科,安徽合肥230000 [2]安徽医科大学,安徽合肥230032 [3]安徽医科大学第二附属医院药物临床实验研究中心,安徽合肥230000

出　　处：《中国药理学通报》2025年第1期116-121,共6页Chinese Pharmacological Bulletin

基　　金：安徽医科大学研究生科研与实践创新项目(No YJS20230196);安徽省转化医学研究(No 2022zhyx-C39);安徽省高等学校自然科学研究项目(No KJ2021A0317)。

摘　　要：目的分析CYFIP1(cytoplasmic FMR1-interacting protein-1,CYFIP1)在结直肠癌中的表达水平,并探究敲低CYFIP1对结直肠癌细胞HT29增殖与凋亡的影响及其可能机制。方法通过免疫组化实验检测32例结直肠癌组织及对应癌旁组织中CYFIP1的表达水平;通过GEPIA2数据库筛选共表达基因,预测二者相关性及可能结合位点;构建siRNA-CYFIP1后,分别用CCK-8、细胞凋亡荧光Hoechst 33342/PI双染实验和Western blot分别检测HT29细胞的增殖、凋亡水平以及细胞凋亡相关蛋白表达水平的变化。结果免疫组化结果显示结直肠癌组织中CYFIP1的表达水平明显高于其对应的癌旁组织(P<0.05);CYFIP1的表达与患者的年龄、性别无关,与TNM分期,淋巴结转移相关(P<0.05);利用GEPIA2、JASPAR数据库和rVista 2.0启动子预测软件在CYFIP1基因上游的3kbps的DNA区域预测了一个保守的TP53结合位点;HT29转染siRNA-CYFIP1后,细胞增殖能力均明显降低(P<0.05);HT29转染siRNA-CYFIP1后细胞凋亡相关蛋白cleaved caspase-3水平升高,caspase-3,Bcl-2的表达水平降低(P<0.05),这可能与CYFIP1与TP53相互作用有关。结论CYFIP1在结直肠癌中高表达,与TNM分期,以及淋巴结转移相关,敲低CYFIP1后可以抑制结直肠癌细胞HT29的增殖、影响相关凋亡蛋白表达。Aim To investigate the expression levels of cytoplasmic FMR1-interacting protein-1(CYFIP1)in colorectal cancer and assess the impact of CYFIP1 interaction on the proliferation and apoptosis of colorectal cancer cell HT29,along with its potential mechanisms.Methods Immunohistochemistry was employed to assess CYFIP1 expression in 32 colorectal cancer tissues and adjacent tissues.Coexpressed genes were identified using the GEPIA2 website to predict potential correlations and binding sites.Following the construction of a siRNA-CYFIP1,alterations in cell proliferation,apoptosis,and levels of apoptosis-related proteins were evaluated through CCK-8 assay,Hoechst 33342/PI double staining assay,and Western blot analysis,respectively.Results The immunohistochemical findings revealed a significantly elevated level of CYFIP1 expression in colorectal cancer tissues compared to paracancer tissues(P<0.05).The expression of CYFIP1 did not show any correlation with age and gender,but exhibited associations with TNM stage and lymph node metastasis(P<0.05).A conserved TP53 binding site was predicted in the 3kbps DNA region upstream of the CYFIP1 gene using GEPIA2,JASPAR databases,and rVista 2.0 promoter prediction software.Following transfection of HT29 cells with siRNA-CYFIP1,the clonogenesis and proliferation of cells significantly decreased(P<0.05).Additionally,the levels of cleaved caspase-3 were elevated,while the expression levels of caspase-3 and Bcl-2 were reduced after transfection with siRNA-CYFIP1(P<0.05),which might be related to the interaction between CYFIP1 and TP53.Conclusions The upregulation of CYFIP1 in colorectal cancer is associated with TNM stage and lymph node metastasis.Upon silencing,CYFIP1 demonstrates the ability to suppress proliferation in HT29 cells and modulate the expression of apoptotic proteins.

关 键 词：结直肠癌 CYFIP1 凋亡 增殖 HT29 小分子干扰RNA 

分 类 号：R735.34[医药卫生—肿瘤]