Downregulation of MiR-93 Expression Reduces Cell Proliferation and Clonogenicity of HepG2 Cells
Downregulation of MiR-93 Expression Reduces Cell Proliferation and Clonogenicity of HepG2 Cells
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MiR-93 表达下调可减少 HepG2 细胞的细胞增殖和克隆形成
DOI：
10.5754/hge12458
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发表时间：
2012-11-01
影响因子：
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通讯作者：
Lin, Ju-Sheng
中科院分区：
文献类型：
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作者：
Xu, Dong;He, Xing-Xing;Lin, Ju-Sheng
背景/目的：在各种类型的癌症中观察到miR-93。本研究旨在探讨miR-93在肝癌发生中的作用。方法：采用RT-PCR方法检测miR-93在HepG 2细胞和原代人肝细胞（PHHC）中的表达。用miR-93抑制剂或阴性对照转染HepG 2细胞。通过使用CellTiter 96（R）Aqueous One Solution Cell Proliferation Assay试剂盒测定细胞增殖。通过细胞迁移实验、集落形成实验和非贴壁生长实验检测其体外迁移和克隆形成能力。流式细胞仪检测细胞凋亡和细胞周期。RT-PCR和Western blot检测转化生长因子β II型受体（TGF β R 2）和整合素β 8（ITGB 8）的mRNA和蛋白水平。结果如下：与PHHC相比，miR-93在HepG 2细胞中上调，并且miR-93的抑制显著抑制HepG 2细胞的增殖、迁移和集落形成。当miR-93被抑制时，TGFBR 2和ITGB 8的表达上调。结论：我们的研究结果揭示了miR-93在肝癌发生中的重要贡献，并表明TGFBR 2和ITGB 8在此过程中的失调作用。因此，使用miR-93的合成抑制剂可能被证明是一种有希望的肝癌治疗方法。
Background/Aims: MiR-93 was observed in various types of cancers. This study is to investigate a role of miR-93 in the carcinogenesis of HCC. Methodology: The expression of miR-93 in HepG2 cells and primary human hepatocytes (PHHC) was measured by RT-PCR. HepG2 cells were transfected with miR-93 inhibitor or negative control. The cell proliferation was determined by using the CellTiter 96 (R) Aqueous One Solution Cell Proliferation Assay kit. The migration and clonogenicity in vitro were measured by cell migration assay, colony formation analysis and anchorage-independent growth assay. The apoptosis and cell cycle were detected by flow cytometry analysis. The mRNA and protein levels of transforming growth factor-beta type II receptor (TGFBR2) and integrin beta8 (ITGB8) were evaluated by RT-PCR and western blot analysis. Results: MiR-93 was upregulated in HepG2 cells compared with PHHC and inhibition of miR-93 significantly suppressed HepG2 cell proliferation, migration and colony formation. The expressions of TGFBR2 and ITGB8 were upregulated when miR-93 was inhibited. Conclusions: Our results reveal an important contribution for miR-93 in hepatocarcinogenesis and suggest a role for TGFBR2 and ITGB8 dysregulation in this process. Thus, the use of synthetic inhibitor of miR-93 may prove to be a promising approach to liver cancer treatment.