MiR-375 Impairs the Invasive Capabilities of Hepatoma Cells by Targeting HIF1α Under Hypoxia
MiR-375 Impairs the Invasive Capabilities of Hepatoma Cells by Targeting HIF1α Under Hypoxia
复制标题
MiR-375 通过在缺氧下靶向 HIF1α 损害肝癌细胞的侵袭能力
DOI：
10.1007/s10620-020-06202-9
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发表时间：
2020-03-25
影响因子：
3.1
通讯作者：
Chang, Ying
中科院分区：
文献类型：
--
作者：
Wang, Chun;Luo, Jie;Chang, Ying
背景和目的由于肝癌的过度生长和血供不足，缺氧是肝癌最普遍的微环境应激之一。HIF 1 α作为一种重要的转录因子参与了肝癌细胞在缺氧条件下多种生物学行为的调节。我们前期的研究表明miR-375是一种缺氧相关的miRNA。然而，miR-375和HIF 1 α之间的相互作用仍不清楚。方法采用生物信息学方法筛选miRNA。采用qRT-PCR、Western blotting和免疫组化染色检测相关分子的表达。生物信息学分析和双荧光素酶测定用于预测和进一步确认靶缔合。分别采用Transwell小室法和流式细胞术检测肝癌细胞的迁移、侵袭和凋亡。结果MiR-375在人HCC与背景肝（BL）、HCC与正常肝组织（NLT）中的表达存在明显差异。在大鼠模型中，miR-375在肝癌发生过程中逐渐下降。在人HCC与BL、HCC与NLT、BL与NLT以及大鼠纤维化肝脏与NLT中，HIF 1 α在蛋白水平而非mRNA水平显著上调。HIF 1 α被确定为miR-375的靶标。MiR-375抑制剂可诱导肝癌细胞在缺氧条件下的迁移和侵袭能力，并抑制其凋亡。通过siRNA阻断HIF 1 α可部分逆转miR-375抑制剂在缺氧条件下的功能。结论MiR-375可通过靶向HIF 1 α抑制肝癌细胞在缺氧条件下的侵袭能力。
Background and Aims Hypoxia represents one of the most pervasive microenvironmental stresses in HCC due to the overwhelming growth and inadequate blood supply. HIF1 alpha as an important transcription factor participates in the regulation of various biological behaviors of HCC cells under hypoxia. Our previous study indicated that miR-375 is a hypoxia-associated miRNA. However, the interaction between miR-375 and HIF1 alpha remains unclear. Methods Bioinformatic analysis was performed for miRNA screening. qRT-PCR, western blotting, and immunohistochemical staining were used to detect the expression of related molecules. Bioinformatic analysis and dual luciferase assay were used to predict and further confirm the target association. Transwell chamber assay and flow cytometry were, respectively, used to detect migration, invasion and apoptosis of hepatoma cells. Results MiR-375 presented an obviously differential expression in human HCCs versus background livers (BLs) and HCCs versus normal liver tissues (NLTs). In rat models, miR-375 was gradually declined during hepatocarcinogenesis. HIF1 alpha was remarkably upregulated at protein level rather than at mRNA level in human HCCs versus BLs, HCCs versus NLTs, BLs versus NLTs, and in rat fibrotic livers versus NLTs. HIF1 alpha was determined to be a target of miR-375. MiR-375 inhibitor induced the migration and invasive capabilities and attenuated apoptosis of hepatoma cells under hypoxia. Depriving HIF1 alpha by siRNA could partially reverse the function of miR-375 inhibitor under hypoxia. Conclusions MiR-375 impairs the invasive capabilities of HCC cells by targeting HIF1 alpha under hypoxia.