LINC00319通过miR-185-3p/FOXK1轴对乳头状甲状腺癌细胞的作用  

Effect of LINC00319 on papillary thyroid carcinoma cells through the miR-185-3p/FOXK1 axis

作　　者：贺雅静[1] 王延 黄立娜 潘翔珍[1] 张玉文[1] HE Yajing;WANG Yan;HUANG Lina;PAN Xiangzhen;ZHANG Yuwen(Department of Pathology,Shangqiu First People’s Hospital,Shangqiu 476000,China)

机构地区：[1]商丘市第一人民医院病理科,河南商丘476000

出　　处：《青岛大学学报(医学版)》2025年第1期13-18,共6页Journal of Qingdao University(Medical Sciences)

基　　金：河南省医学科技攻关计划项目(LHGJ20-210985)。

摘　　要：目的探讨LINC00319对乳头状甲状腺癌细胞生物学行为的影响及其可能机制。方法应用反转录实时定量PCR(qRT-PCR)与Western blot方法,分别检测乳头状甲状腺癌组织和细胞LINC00319、miR-185-3p、叉头框转录因子1(FOXK1)的表达。将si-NC、si-LINC00319、miR-NC、miR-185-3p mimics、si-NC、si-FOXK1、pcDNA+si-LINC00319、pcDNA-FOXK1+si-LINC00319分别转染至甲状腺癌BCPAP细胞,通过双荧光素酶报告实验验证miR-185-3p、LINC00319及FOXK1的靶向关系;应用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)、平板克隆形成实验、流式细胞术和Transwell实验检测BCPAP细胞增殖、凋亡、迁移和侵袭;采用Western blot方法检测Cleaved-caspase-3、基质金属蛋白酶2(MMP2)、基质金属蛋白酶9(MMP9)蛋白表达。结果与癌旁组织和人正常甲状腺细胞比较,乳头状甲状腺癌组织及其细胞系中LINC00319、FOXK1表达升高(t=27.19~39.06,F=96.94~216.25,P<0.05),miR-185-3p表达降低(t=21.83,F=216.25,P<0.05)。LINC00319可靶向miR-185-3p,miR-185-3p可靶向FOXK1。与相应阴性对照相比,转染si-LINC00319、miR-185-3p mimics及si-FOXK1均可以抑制细胞活力、MMP2表达、MMP9表达、细胞克隆形成、迁移和侵袭细胞数(F=121.22~180.59,t=12.48~22.59,P<0.05),提高细胞凋亡率和Cleaved-caspase-3蛋白表达(F=586.51、196.33,t=16.43~21.61,P<0.05);共转染si-LINC00319与pcDNA-FOXK1可以恢复si-LINC00319对BCPAP细胞生物学行为的作用。结论LINC00319可通过靶向miR-185-3p/FOXK1轴促进乳头状甲状腺癌细胞的恶性行为。Objective To investigate the effect of LINC00319 on the biological behavior of papillary thyroid carcinoma cells and its possible mechanism.Methods The methods of qRT-PCR and Western blot were used to measure the expression of LINC00319,miR-185-3p,and FOXK1 in papillary thyroid carcinoma tissue and cells.Thyroid carcinoma BCPAP cells were transfected with si-NC,si-LINC00319,miR-NC,miR-185-3p mimics,si-NC,si-FOXK1,pcDNA+si-LINC00319,and pcDNA-FOXK1+si-LINC00319,respectively,and dual luciferase reporter assay was used to validate the targeting relationship between miR-185-3p,LINC00319,and FOXK1.MTT assay,plate colony formation assay,flow cytometry,and Transwell assay were used to measure the proliferation,apoptosis,migration,and invasion of BCPAP cells,and Western blot was used to measure the protein expression levels of Cleaved caspase-3,matrix metalloproteinase-2(MMP2),and matrix metalloproteinase-9(MMP9).Results Compared with adjacent tissue and normal human thyroid cells,papillary thyroid carcinoma tissue and cells showed significant increases in the expression of LINC00319 and FOXK1(t=27.19-39.06,F=96.94-216.25,P<0.05)and a significant reduction in the expression of miR-185-3p(t=21.83,F=216.25,P<0.05).LINC00319 could target miR-185-3p,and miR-185-3p could target FOXK1.Compared with the corresponding negative control group,transfection with si-LINC00319,miR-185-3p mimics,or si-FOXK1 reduced cell viability,the expression of MMP2 and MMP9,the number of clones formed,and the number of migration and invasion cells(F=121.22-180.59,t=12.48-22.59,P<0.05),while it increased cell apoptosis rate and the expression of Cleaved caspase-3(F=586.51,196.33;t=16.43-21.61;P<0.05).Co-transfection with si-LINC00319 and pcDNA-FOXK1 could restore the effect of si-LINC00319 on the biological behavior of BCPAP cells.Conclusion LINC00319 can promote the malignant beha-vior of papillary thyroid carcinoma cells by targeting the miR-185-3p/FOXK1 axis.

关 键 词：甲状腺癌 乳头状 LINC00319 细胞增殖 分子生物学 

分 类 号：R736.1[医药卫生—肿瘤]