首页|长链非编码RNA CASC15对肝细胞癌增殖、侵袭和迁移的影响及作用机制

长链非编码RNA CASC15对肝细胞癌增殖、侵袭和迁移的影响及作用机制

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目的 探究长链非编码RNA(lncRNA)癌症易感性基因 15(Cancer susceptibility candidate 15,CASC15)在肝细胞癌(Hepatocellular carcinoma,HCC)中的分子调控机制.方法 通过生物信息学方法预测目的基因表达,并分析目的基因表达与患者生存时间的关系;收集临床HCC患者肝癌组织和癌旁组织;CCK-8、Transwell和流式细胞术实验检测HCC细胞SMMC7721 和Huh-7 的增殖、侵袭、迁移以及凋亡;双荧光素酶实验检测miR-144-3p与CASC15 和富含亮氨酸的重复序列蛋白 1(Leucine rich repeat containing protein 1,LRRC1)的靶向关系;qRT-PCR和Western blot检测mRNA和蛋白表达情况;免疫荧光实验用于蛋白定位研究;回复实验验证CASC15/miR-144-3p/LRRC1 对HCC进展的影响.体内实验验证CASC15 对HCC进展的影响.结果 TCGA数据库与qRT-PCR检测显示HCC组织和细胞中CASC15 高表达、miR-144-3p低表达、LR-RC1 高表达(P<0.05).增殖、侵袭、迁移的细胞功能实验结果表明在肿瘤发展中CASC15 和LRRC1 起到促进作用,miR-144-3p则是抑制作用,与凋亡实验结果一致(P<0.05).细胞功能实验表明CASC15 抑制miR-144-3p功能,miR-144-3p抑制LRRC1,CASC15 与miR-144-3p结合,并导致LRRC1 上调.回复实验结果表明CASC15 通过抑制miR-144-3p促进LRRC1的表达从而促进HCC细胞增殖、侵袭和迁移并抑制细胞凋亡.结论 CASC15 可能通过调节miR-144-3p/LRRC1 轴,从而促进HCC进展.
The effects and mechanism of long non-coding RNA CASC15 on proliferation,invasion and migration of hepatocellular car-cinoma
Objective The aim of this study was to investigate the molecular regulatory mechanism of cancer susceptibility candidate 15(CASC15),a long-stranded non-coding RNA(lncRNA),in hepatocellular carcinoma(HCC).Methods Bioinformat-ics methods were used to predict the expression of target genes and analyze the relationship between the expression of target genes and the survival time of patients;Hepatocellular carcinoma tissues and adjacent tissues from patients with HCC were collected;CCK-8,Tr-answell,and flow cytometry experiments were used to detect proliferation,invasion,migration and apoptosis of SMMC7721 cells and Huh-7 cells;The dual-luciferase assay was used to detect the targeting relationship between miR-144-3p and CASC15,as well as leucine rich repeat containing protein 1(LRRC1);RT-qPCR and Western blot were used to detect mRNA and protein expression of target genes;Immunofluorescence was used for protein localization of target genes;Replicate experiment was performed to verify the effect of CASC15/miR-144-3p/LRRC1 on the progression of HCC.In vivo experiment was performed to verify the effect of CASC15 on HCC progression.Results TCGA database and RT-qPCR assay showed high expression of CASC15,low expression of miR-144-3p,and high expression of LRRC1 in HCC tissues and cells(P<0.05).The results of cell function experiments on proliferation,inva-sion and migration showed that CASC15 and LRRC1 played a promoting role in tumor development,while miR-144-3p had an inhibi-tory effect,consistent with the results of apoptosis experiments(P<0.05).Cell function experiments showed that CASC15 inhibited miR-144-3p function,miR-144-3p inhibited LRRC1,and CASC15 bound to miR-144-3p,leading to the upregulation of LRRC1.The replicate experimental results indicated that CASC15 promoted LRRC1 expression through inhibiting miR-144-3p,thereby pro-moting HCC cell proliferation,invasion and migration,and inhibiting apoptosis.Conclusion CASC15 may promote HCC progression by regulating the miR-144-3p/LRRC1 axis.

Cancer susceptibility gene 15miR-144-3pLeucine-rich repeative sequence protein 1Hepatocellular carcino-ma cellsCell biological function

吴东洋、蔡青山、刘东、左刚刚、李树栋、刘立友、郑建兴

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唐山中心医院肝胆外科(唐山 063000)

癌症易感性基因15 miR-144-3p 富含亮氨酸的重复序列蛋白1 肝癌细胞 细胞生物学功能

河北省医学科学研究项目

20221840

2024

实用肿瘤学杂志
黑龙江省,辽宁省,吉林省肿瘤防治办公室

实用肿瘤学杂志

CSTPCD
影响因子:0.528
ISSN:1002-3070
年,卷(期):2024.38(1)
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