LINC01915对人结直肠癌细胞增殖、迁移、侵袭的作用及机制OA
Effects of LINC01915 on proliferation,migration,and invasion of human colorectal cancer cells and its mechanism
目的 观察LINC01915对人结直肠癌细胞增殖、迁移、侵袭的作用,并探讨其可能的调控机制.方法 取人结直肠癌细胞系SW620、SW480、LOVO、HCT116与正常人结直肠细胞系NCM460,RT-qPCR测定LINC01915表达.取LINC01915表达最高的HCT116细胞系,分为上调组、下调组、上调对照组、下调对照组、空白组.荧光显微镜检测各组细胞转染效率;RT-qPCR检测各组细胞LINC01915表达;二苯基四氮唑溴盐(MTT)法检测各组细胞增殖活性;划痕愈合实验和Transwell检测各组细胞迁移、侵袭活性;RT-qPCR检测细胞miR-92a-3p及大肿瘤抑制激酶2(LATS2)、转录因子21(TCF21)、Kruppel样因子4(KLF4)mRNA表达;Western blot检测细胞LATS2、TCF21、KLF4蛋白表达.双荧光素酶报告基因实验验证LINC01915与miR-92a-3p的靶向关系.结果 各种人结直肠癌细胞系LINC01915表达均低于NCM460细胞(P<0.05),且癌细胞系中HCT116的LINC01915表达最高(P<0.05);各转染组细胞转染效率均较高;与空白组、上调对照组比较,上调组细胞LINC01915表达及LATS2、TCF21、KLF4 mRNA与蛋白表达均升高(P<0.05),A值、划痕愈合率、侵袭细胞数、miR-92a-3p表达均下降(P<0.05);与空白组、下调对照组比较,下调组细胞中LINC01915表达及LATS2、TCF21、KLF4 mRNA与蛋白表达均下降(P<0.05),A值、划痕愈合率、侵袭细胞数、miR-92a-3p表达均升高(P<0.05).LINC01915与miR-92a-3p存在结合位点,且与miR-NC组相比,miR-92a-3p mimics组WT-LINC01915荧光素酶活性降低(P<0.05).结论 LINC01915在人结直肠癌细胞中低表达,上调LINC01915表达可抑制细胞的增殖、迁移和侵袭能力,这可能与抑制miR-92a-3p上调LATS2、TCF21、KLF4表达有关.
Objective To observe the effects of LINC01915 on the proliferation,migration and invasion of human colorectal cancer cells,and to explore its possible regulatory mechanism.Methods The colorectal cancer cell lines of SW620,SW480,LOVO,HCT116 and the normal colorectal cell line of NCM460 were selected,and the expression of LINC01915 was detected by RT-qPCR.HCT116 cell line with the highest expression of LINC01915 was taken and divided into the upregulation group,the downregulation group,the upregulation control group,the downregulation control group and the blank group.The transfection efficiency of each group was detected by fluorescence microscope;RT-qPCR was used to detect the expression of LINC01915 in each group;methyl thiazolyl tetrazolium(MTT)assay was used to detect the proliferative activity in each group;scratch wound healing assay and Transwell assay were used to detect the migration and invasion activities in each group;RT-qPCR was used to detect the expression of miR-92a-3p and mRNA expression of large tumor suppressor homolog 2(LATS2),transcription factor 21(TCF21)and Kruppel like factor 4(KLF4);and Western blot was used to detect the expression of LATS2,TCF21,and KLF4 proteins.Dual-fluorescein reporter assay was used to verify the targeting relationship between LINC01915 and miR-92a-3p.Results The expression of LINC01915 in various human colorectal cancer cell lines were lower than that in the NCM460 cell(P<0.05),and the highest LINC01915 expression in human colorectal cancer cell lines was observed in HCT116(P<0.05).The transfection efficiency of cells in each transfection group was high.Compared with the blank group and the upregulation control group,the expression of LINC01915,and mRNA and protein expression of LATS2,TCF21,and KLF4 in the upregulation group increased(P<0.05),and the A value,scratch healing rate,number of invasive cells and miR-92a-3p expression decreased(P<0.05).Compared with the blank group and the downregulation control group,the expression of LINC01915,and mRNA and protein expression of LATS2,TCF21,and KLF4 in the downregulation group decreased(P<0.05),and the A value,scratch healing rate,number of invasive cells and miR-92a-3p expression increased(P<0.05).LINC01915 had binding sites with miR-92a-3p,and compared with the miR-NC group,the miR-92a-3p mimics group showed a decrease in the luciferase activity of WT-LINC01915(P<0.05).Conclusion The expression of LINC01915 in the human colorectal cancer cell lines decreases,and upregulation of LINC01915 expression can inhibit cell proliferation,migration,and invasion,which may be related to the up-regulation of the expression of LATS2,TCF21 and KLF4 by inhibiting miR-92a-3p.
韩炜;李程;李文翰;霍斌亮;师文
陕西省人民医院肿瘤外科,陕西 西安,710068陕西省人民医院肿瘤外科,陕西 西安,710068陕西省人民医院肿瘤外科,陕西 西安,710068陕西省人民医院肿瘤外科,陕西 西安,710068西安交通大学第一附属医院消化内科,陕西 西安 710061
临床医学
LINC01915结直肠癌增殖迁移侵袭
LINC01915colorectal cancerproliferationmigrationinvasion
《局解手术学杂志》 2025 (4)
295-300,6
陕西省自然科学基础研究计划(2021JQ-917)陕西省人民医院科技人才支持计划(2021JY-46)
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