DTX2促进奥沙利铂耐药的结直肠癌细胞增殖、侵袭和上皮间质转化  

High expression of DTX2 promotes proliferation,invasion and epithelial-mesenchymal transition of oxaliplatin-resistant colorectal cancer cells

作　　者：马振南 刘福全[1] 赵雪峰[1] 张晓微[1] MA Zhennan;LIU Fuquan;ZHAO Xuefeng;ZHANG Xiaowei(Department of Anorectal Surgery,Dalian University Affiliated Xinhua Hospital,Dalian 116021,China)

机构地区：[1]大连大学附属新华医院肛肠外科,辽宁大连116021

出　　处：《南方医科大学学报》2025年第4期829-836,共8页Journal of Southern Medical University

基　　金：大连市生命健康领域指导计划项目(2023005).

摘　　要：目的探讨DTX2对奥沙利铂耐药的结直肠癌(CRC/OXA)细胞的影响及作用机制。方法利用CCK8检测奥沙利铂(OXA)对CRC细胞的抑制率,构建CRC/OXA细胞系,检测CRC/OXA细胞中DTX2的表达水平,利用基因工具干预CRC/OXA细胞,分为未转染组(con)、敲低组(DTX2-shRNA)及共转染组(DTX2-shRNA+pcDNA-Notch2)。采用平板克隆、划痕和Transwell侵袭实验检测改变DTX2的表达对CRC/OXA细胞增值、迁移侵袭能力的影响,并通过Western blotting检测各组中Notch2、NICD、Ecadherin、N-cadherin及Vimentin蛋白的表达水平。利用SW620/OXA细胞同样分组行裸鼠移植瘤实验,体内验证对裸鼠成瘤及蛋白的影响。结果OXA对CRC细胞有明显抑制作用,SW620和LoVo细胞IC50分别为6.00和8.00μmol/L,成功构建CRC/OXA细胞系,CRC/OXA细胞中DTX2表达量明显升高(P<0.01)。DTX2-shRNA组中CRC/OXA细胞明显抑制增值、迁移侵袭能力(P<0.05),DTX2-shRNA+pcDNA-Notch2组可逆转增强CRC/OXA细胞增值、迁移侵袭的能力(P<0.05)。Notch2、NICD及Vimentin蛋白平均表达水平,在DTX2-shRNA组中明显降低,而DTX2-shRNA+pcDNA-Notch2组明显升高(P<0.01);E-cadherin蛋白表达水平在以上两组中的趋势相反且有明显差异(P<0.01)。体内实验显示DTX2可明显促进SW620/OXA细胞移植瘤的生长及对应蛋白的变化(P<0.05)。结论DTX2通过Notch2信号通路促进CRC/OXA细胞增值、迁移侵袭及上皮间质转化,DTX2可能作为提高OXA疗效的分子标志物。Objective To investigate the role of DTX2 in regulating biological behaviors of oxaliplatin-resistant colorectal cancer cells(CRC/OXA cells).Methods CCK8 assay was used to determine the inhibition rate of oxaliplatin-treated CRC cells.A CRC/OXA cell line was constructed,in which DTX2 expression level was detected.The cells were transfected with a DTX2-shRNA plasmid or co-transfected with DTX2-shRNA and pcDNA-Notch2,and the changes in cell proliferation,migration and invasion ability were evaluated using plate cloning assay,scratch assay and Transwell invasion assay.The expression levels of Notch2,NICD and epithelial-mesenchymal transition(EMT)proteins of the transfected cells were detected with Western blotting.In a nude mouse model bearing SW620/OXA cell xenografts,the effects of DTX2 knockdown and Notch2 overexpression in the implanted cells on tumor growth and protein expressions were tested.Results The IC50 of oxaliplatin was 6.00μmol/L in SW620 cells and 8.00μmol/L in LoVo cells.CRC/OXA cells showed a significantly increased expression of DTX2.DTX2 knockdown in CRC/OXA cells significantly inhibited cell proliferation,migration and invasion,and these effects were reversed by co-transfection of the cells with pcDNA-Notch2.DTX2 knockdown significantly reduced the expression levels of Notch2,NICD and vimentin proteins and increased E-cadherin expression in CRC/OXA cells,and co-transfection with pcDNA-Notch2 potently attenuated the changes in these proteins.In the tumor-bearing mice,DTX2 overexpression obviously promoted the growth of SW620/OXA cell xenograft,enhanced the protein expressions of Notch2,NICD and vimentin,and lowered the expression of E-cadherin.Conclusion High expression of DTX2 promotes proliferation,migration,invasion and EMT of CRC/OXA cells through the Notch2 signaling pathway,suggesting the potential of DTX2 as a target to improve the efficacy of oxaliplatin.

关 键 词：deltex-2 结直肠癌 Notch2信号通路 上皮间质转化 奥沙利铂 耐药 

分 类 号：R735.3[医药卫生—肿瘤]