摘要
目的 探讨hsa-miR-374a-3p与VEGFB在结直肠癌中的相关性和作用。方法 TCGA数据集分析VEGFB在结直肠癌中的表达和对生存曲线的影响;免疫组织化学方法检测59例结直肠癌组织中VEGFB蛋白的表达;ELISA方法检测结直肠正常细胞和结直肠癌细胞中VEGFB的表达。重组人VEGFB蛋白或VEGFB抗体处理后,MTT法检测细胞活力;膜联蛋白V-PE和7-氨基放线菌素D(7-AAD)染色法检测细胞凋亡水平;FASAria细胞分选仪检测细胞周期。miRNA测序检测细胞中miRNA总体表达水平;实时荧光定量PCR检测miRNA-374a-3p与VEGFB mRNA的表达水平;荧光素酶报告实验验证miRNA-374a-3p与VEGFB-3′-UTR的靶向关系。结果 VEGFB在结直肠癌组织中的mRNA和蛋白表达水平显著高于正常组织。VEGFB高表达患者的中位生存期明显低于低表达患者。与结直肠正常细胞相比,结直肠癌细胞VEGFB表达水平明显升高。VEGFB蛋白处理后,结直肠正常细胞NCM460的细胞活力水平上升、细胞凋亡水平下降。结直肠癌组织中miR-374a-3p与VEGFB的表达水平存在负相关。miR-374a-3p靶向VEGFB mRNA的3′端非编码区。敲低miR-374a-3p后,细胞中VEGFB的mRNA和蛋白表达水平均上升,细胞活力上升,细胞凋亡减少。结论 miR-374a-3p与VEGFB的表达水平在结直肠癌中存在负相关,VEGFB/miR-374a-3p轴调控结直肠癌细胞的增殖与凋亡。
Objective To investigate the correlation and role of hsa-miR-374a-3p and VEGFB(Vascular endothelial growth factor B)in colorectal cancer.Methods The TCGA dataset was used to analyze the expression of VEGFB in colorectal cancer and its effect on survival curve.The expression of VEGFB in 59 cases of colorectal cancer was detected by immunohistochemistry.The expression of VECFB in normal colorectal and colorectal cancer cells was detected by ELISA.After treatment with recombinant human VEGFB protein or VEGFB antibody,the cell viability was detected by MTT assay.Cell apoptosis was detected by annexin V-PE and 7-amino-actinomycin D(7-AAD)staining.Cell cycle was measured with FASAria cell sorter.The overall expression level of miRNA was detected by miRNA sequencing.The mRNA levels of miRNA-374a-3p and VEGFB were detected by real-time fluorescence quantitative PCR.Luciferase reports experiments to verify the targeting relationship between miRNA-374a-3p and VEGFB-3'-UTR.Results The mRNA and protein levels of VEGFB in colorectal cancer tissues were significantly higher than those in normal tissues.Patients with high VEGFB expression had significantly lower median survival than those with low VEGFB expression.The expression of VEGFB in colorectal cancer cells was significantly higher than that in normal colorectal cancer cells.After VEGFB protein treatment,the viability of NCM460 cells was increased and the apoptosis of cells was decreased.The level of miR-374a-3p was negatively correlated with VEGFB in colorectal cancer tissues.miR-374a-3p targets the 3'end non-coding region of VEGFB mRNA.After the knockdown of miR-374a-3p,the mRNA and protein levels of VEGFB in the cells were increased,the vitality of cells was increased,and the apoptosis of cells was decreased.Conclusion The levels of miR-374a-3p and VEGFB are negatively correlated in colorectal cancer,and the VEGFB/miR-374a-3p axis regulates the proliferation and apoptosis of colorectal cancer cells.
作者
朱亚雯
达静
徐凤霞
姜树中
陆翠华
ZHU Ya-wen;DA Jing;XU Feng-xia;JIANG Shu-zhong;LU Cui-hua(Department of Gastroenterology,Nantong Sixth People's Hospital,Nantong 226001;Department of Gastroenterology,Affiliated Hospital of Nantong University,Nantong 226006,China)
出处
《解剖科学进展》
CAS
2023年第3期295-298,共4页
Progress of Anatomical Sciences
基金
南通市卫生健康委员会项目(MB2021071)。
作者简介
通信作者:陆翠华。
