MiR-150-5p靶向TP53对结直肠癌细胞侵袭和增殖的影响

MiR-150-5p Promotes Invasion and Proliferation by Targeting TP53 in Colorectal Cancer Cells

目的:探索miR-150-5p靶向调控TP53基因对结直肠癌(colorectal cancer,CRC)在临床和生物学的相关性,研究miR-150-5p调控TP53基因在结直肠癌细胞增殖和侵袭病变中的作用。方法:收集临床手术切除并病理证实的结直肠癌患者的手术及血浆样本(结直肠癌和癌旁组织)60例,另选取癌旁正常粘膜10例,腺瘤30例。根据瘤体直径分为肿瘤>5 cm(n=30)和≤5 cm(n=30)。q RT-PCR法测定样本中miR-150-5p表达,荧光素酶活性测定以确定TP53是否为miR-150-5p的靶基因。使用SW480细胞株,进行Transwell小室检测细胞侵袭能力,CCK-8检测细胞增值能力,对miR-150-5p进行生物信息学分析。结果:TP53是miR-150-5p的下游基因。癌组织及血浆中miR-150-5p表达量低于癌旁组织,直径>5 cm瘤体中的miR-150-5p表达量显著低于直径≤5 cm瘤体,Ⅰ-Ⅱ期结直肠癌组织中的miR-150-5p表达显著高于Ⅲ-Ⅳ期(P<0.05)。上调miR-150-5p后,细胞中TP53表达下降,下调miR-150-5p后,TP53表达升高;CCK-8增殖试验显示细胞中miR-150-5p过表达组抑制细胞增殖(P<0.05)。结论:Mi R-150-5p在结直肠癌组织和细胞中显著低表达,miR-150-5p通过靶向调节TP53抑制人CRC细胞的侵袭和增殖,有望成为结直肠癌治疗的新靶点。

Objective:To explore the clinical and biological relevance of miR-150-5p targeted regulation of TP53 gene in colorectal cancer,and to investigate the role of miR-150-5p regulates TP53 gene on behavior of invasion and proliferation in colorectal cancer cells.Methods:The surgical and plasma samples(colorectal cancer and paracancerosal tissue)of 60 patients with colorectal cancer who had undergone surgical resection and pathologically confirmed colorectal cancer were collected.There were 10 normal mucosa and 30 adenomas.According to tumor diameter,tumors were divided into tumors>5 cm(n=30)and≤5 cm(n=30).qRT-PCR was used to determine the expression of miR-150-5p in the samples,and luciferase activity was performed to determine whether TP53 was the target gene of miR-150-5p.Using human colorectal cancer cell line SW480,Transwell chamber was used to detect cell invasion ability,CCK-8 was used to detect cell value-added ability,and bioinformatics analysis was performed on miR-150-5p.Results:TP53 was the target gene of miR-150-5p.The expression of miR-150-5p in cancer tissues and plasma was lower than that in adjacent tissues,the expression of miR-150-5p in tumors with a diameter of>5 cm was significantly lower than that of tumors with a diameter of≤5 cm,miR-150-5p expression in stage I-II colorectal cancer tissues was significantly higher than that in stage II-IV tissue(P<0.05).Afer up-regulating miR-150-5p,TP53 expression in the cells decreased,and after down-regulating miR-150-5p,TP53 expression increased;CCK-8 proliferation test showed that miR-150-5p over-expression group inhibited cell proliferation(P<0.05).Conclusion:MiR-150-5p is significantly under-expressed in colorectal cancer tissues and cells.MiR-150-5p inhibits the invasion and proliferation of human CRC cells by targeting TP53,and is expected to become a new target for colorectal cancer treatment.