摘要
目的 探讨TRIM59促进结直肠癌进展的分子机制。方法 收集28例行手术治疗的人结直肠肿瘤组织样本及临床资料,采用免疫荧光双染技术分析癌与癌周正常组织样本肿瘤相关巨噬细胞中TRIM59的表达,应用流式分选技术分选出CD68^(+)的肿瘤相关巨噬细胞,Western blot方法检测TRIM59的表达。培养人巨噬细胞THP-1细胞系,随机分为sh-TRIM59组和NC组,分别转染sh-TRIM59以及阴性对照,并与人结肠癌细胞SW480进行共培养,应用Western blot方法检测两组THP-1细胞中PI3K/AKT/mTOR通路相关蛋白p-PI3K、p-AKT、p-mTOR及TRIM59、VEGF、COX-2、MMP-9蛋白的表达情况;应用划痕实验检测SW480细胞的迁移能力;应用Transwell检测SW480细胞的侵袭和迁移能力。将HUVEC细胞与上述两种细胞共培养进行血管形成实验,观察两组细胞血管形成能力。结果 免疫荧光实验结果显示:结直肠癌组织肿瘤相关巨噬细胞中TRIM59的表达明显高于癌周正常组织(P<0.01);Western blot结果显示:与癌周正常组织相比,癌组织中肿瘤相关巨噬细胞TRIM59的表达明显升高(P<0.01)。与NC组相比,sh-TRIM59组THP-1细胞中PI3K/AKT/mTOR通路相关蛋白p-PI3K、p-AKT、p-mTOR及TRIM59、COX-2、VEGF、MMP-9表达明显降低(P<0.01);与NC组相比,sh-TRIM59组SW480细胞划痕距离明显增大,细胞愈合率明显减小(P<0.01);与NC组相比,sh-TRIM59组HUVEC细胞新生血管数量明显增加(P<0.01)。结论 TRIM59通过激活结直肠癌肿瘤相关巨噬细胞中PI3K/AKT/mTOR通路促进COX-2、VEGF、MMP-9的分泌,增强癌细胞的浸润、侵袭和内皮细胞的血管生成能力,从而加速肿瘤的进展。
Objective To investigate the molecular mechanism of TRIM59 in promoting the progression of colorectal cancer.Methods The tumor tissue samples and the clinical data of 28 patients with colorectal cancer were collected.The expression of TRIM59 in tumor-associated macrophages in cancer tissues and normal tissues was analyzed by double immunofluorescence staining technique,the CD68^(+) tumor-associated macrophages were separated by flow cytometry and the expression of TRIM59 was detected by Western blot.Human macrophage THP-1 cell line was cultured and randomly divided into sh-TRIM59 group and NC group.The cells were respectively transfected with sh-TRIM59 and negative control,and then co-cultured with SW480 cells.The expression levels of PI3K/AKT/mTOR pathway-related proteins p-PI3K,p-AKT,p-mTOR and TRIM59,VEGF,COX-2,MMP-9 in THP-1 cells were detected by Western blot.The migration ability of SW480 cells was detected by scratch wound assay,and the invasion and migration abilities of SW480 cells were detected by Transwell.HUVEC cells were co-cultured with the above two kinds of cells to observe their ability of angiogenesis by the tube-formation assay.Results The results of double immunofluorescence staining assay showed that the expression of TRIM59 in the tumor-associated macrophages in colorectal cancer tissues was significantly higher than that in the normal tissues around cancer(P<0.01).Western blot results showed that the expression of TRIM59 in the tumor-associated macrophages in cancer tissues was significantly higher than that in normal tissues around cancer(P<0.01).Compared with NC group,the expression levels of PI3K/AKT/mTOR pathway-related proteins p-PI3K,p-AKT,p-mTOR,TRIM59,COX-2,VEGF and MMP-9 in THP-1 cells were significantly decreased in sh-TRIM59 group(P<0.01).Moreover,compared with NC group,the scratch distance of SW480 cells was significantly increased in sh-TRIM59 group,and the cell healing rate was significantly reduced(P<0.01).The number of new blood vessels in HUVEC cells was significantly incre
作者
高晓斌
王胜杰
梁峰
王文静
孙光源
武雪亮
GAO Xiaobin;WANG Shengjie;LIANG Feng;WANG Wenjing;SUN Guangyuan;WU Xueliang(Department of General Surgery,First Hospital Affiliated to Hebei North College,Zhangjiakou 075000,China)
出处
《山西医科大学学报》
CAS
2023年第7期903-909,共7页
Journal of Shanxi Medical University
基金
河北省卫健委医学科学研究课题(20200515)。
