孙波,周雷,周东亚,王小龙,徐超,吕莹,刘犇
SUN Bo,ZHOU Lei,ZHOU Dongya,WANG Xiaolong,XU Chao,LV Ying,LIU Ben

• 1:南京中医药大学沭阳附属医院肿瘤科

摘要(Abstract)：

目的 探讨LncRNA NNT-AS1通过miR-582-5p/MCL1轴对肝癌的侵袭和转移的影响。方法 培养正常肝L-02细胞和肝癌细胞MHCC97H、LM3、SMCC7721、Huh7、Hep3B,选取肝癌Huh7细胞系，将细胞进行分组与转染；采用qRT-PCR检测各组细胞中NNT-AS1、miR-582-5p表达，WB检测各组细胞MCL1蛋白表达，RIP检测LncRNA NNT-AS1和miR-582-5p的结合情况，双荧光素酶报告基因实验验证miR-582-5p和MCL1靶向关系，Transwell实验检测细胞侵袭和转移能力。结果 与正常肝细胞L-02比较，NNT-AS1在肝癌细胞系中高表达，miR-582-5p在肝癌细胞系中低表达；NNT-AS1和miR-582-5p沉默后，细胞中NNT-AS1和miR-582-5p表达显著降低，细胞侵袭和转移能力也明显减弱；LncRNA NNT-AS1可结合吸附miR-582-5p, MCL1是miR-582-5p的下游靶基因、且MCL1在肝癌细胞系中高表达；干扰LncRNA NNT-AS1或过表达miR-582-5p表达可抑制肝癌细胞侵袭和迁移；同时沉默NNT-AS1和过表达miR-582-5p,可显著降低细胞侵袭和迁移能力；干扰MCL1也可抑制肝癌细胞侵袭和迁移能力增强的现象。结论 沉默LncRNA NNT-AS1可通过miR-582-5p作用，抑制MCL1表达从而抑制肝癌细胞侵袭和转移。
Objective To investigate the effect of LncRNA NT-AS1 on invasion and metastasis of liver cancer through miR-582-5p/MCL1 axis. Methods Normal liver cells L-02 cells and liver cancer cells MHCC97H, LM3, SMCC7721, Huh7, and Hep3B were cultured. The Huh7 cell line of liver cancer was selected, and such cells were grouped and transfected. The expressions of NNT-AS1 and miR-582-5p in each group were detected by qRT-PCR. The expression of MCL1 protein in each group was detected by WB. The binding of LncRNA NNT-AS1 and miR-582-5p was detected by RIP. The targeting relationship between miR-582-5p and MCL1 was verified by dual luciferase reporter gene assay. Transwell assay was employed to detect cell invasion and metastasis. Results Compared with normal liver cell L-02, NNT-AS1 was highly expressed in liver cancer cell lines, and miR-582-5p was lowly expressed in liver cancer cell lines. After the silence of NNT-AS1 and miR-582-5p, the expression of NNT-AS1 and miR-582-5p were significantly decreased, and the cell invasion and metastasis ability was also significantly weakened. LncRNA NNT-AS1 could bind and adsorb miR-582-5p. MCL1 was a downstream target gene of miR-582-5p, and MCL1 was highly expressed in liver cancer cell lines. Interfering with LncRNA NNT-AS1 or overexpressing of miR-582-5p could inhibit the invasion and migration of liver cancer cells. Simultaneous silencing of NNT-AS1 and overexpressing of miR-582-5p could significantly reduce cell invasion and migration. Interfering with MCL1 could also inhibit the invasion and migration of liver cancer cells. Conclusion Silencing LncRNA NNT-AS1 can inhibit the expression of MCL1 through miR-582-5p, thereby inhibiting the invasion and metastasis of liver cancer cells.

关键词(KeyWords)： NNT反义核糖核酸1;微小核糖核酸582-5p;髓细胞白血病1;肝肿瘤;侵袭;转移
NNT-antisense RNA1;microRNA-582-5p;myeloid cell leukemia 1;liver cancer;invasion;metastasis

Abstract:

Keywords:

基金项目(Foundation): 国家中医药管理局重点研究室开放课题(202135);; 宿迁市科技计划项目(Z2018020)

作者(Author): 孙波,周雷,周东亚,王小龙,徐超,吕莹,刘犇
SUN Bo,ZHOU Lei,ZHOU Dongya,WANG Xiaolong,XU Chao,LV Ying,LIU Ben

DOI: 10.19367/j.cnki.2096-8388.2023.07.007

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