Department of Traditional Chinese Medicine, JiNing No. 1 People's Hospital, JiNing, China.
Eur Rev Med Pharmacol Sci. 2019 Apr;23(7):2777-2785. doi: 10.26355/eurrev_201904_17551.
Hepatocellular carcinoma (HCC) is a malignant cancer with a high fatality rate, and the expression of microRNA-145 (miR-145) is significantly low in HCC tissue. Therefore, the effect of miR-145 on HCC was explored.
Primary hepatocellular carcinoma samples and corresponding normal samples, and HepG2 cells were analyzed using flow cytometry, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, Real-time quantitative reverse transcription-polymerase chain reaction, Western blotting, and dual-luciferase reporter assay.
miR-145 expression was significantly downregulated in HCC tissue and HepG2 cells as compared to normal liver tissue. After HepG2 cells were transfected with miR-145 mimics, miR-145 expression was recovered, accompanied by a significantly lower cell number, inhibition of the G1/S phase transition, and promotion of the apoptosis of HepG2 cells, as well as changes in levels of G1/S-specific cyclin-E1 (CCNE1) and activated caspase-3. Furthermore, the rho-associated protein kinase 1 (ROCK1) levels were opposite the levels of miR-145 expression in vivo and in vitro, and additional experiments with co-transfection of miR-145 mimics and pEGFP-N3-3'UTR provided the direct evidence that the ROCK1 gene is a target of miR-145. Moreover, a significant decrease or increase in the expression of ROCK1 was associated with nuclear factor-kB (NF-κB)(p65) activity, and lipopolysaccharide (LPS) significantly increased NF-κB(p65) activity, accompanied by recovery of the reduction in the number of HepG2 cells for miR-145 mimics. The NF-κB activity and cell number were significantly (p < 0.05, p < 0.01) increased in response to the overexpression of the ROCK1 gene in HepG2 cells.
We showed that miR-145 can target and downregulate ROCK1 expression, and it controls HCC by inhibiting the cell cycle and activating apoptosis via the ROCK1/NF-κB signaling pathway. Our findings will provide a new perspective for the therapy of HCC.
肝细胞癌（HCC）是一种死亡率很高的恶性肿瘤，miR-145 在 HCC 组织中的表达明显降低。因此，探讨了 miR-145 对 HCC 的影响。
分析原发性肝癌组织和相应的正常组织及 HepG2 细胞的流式细胞术、3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）检测、实时定量逆转录-聚合酶链反应、Western blot 及双荧光素酶报告基因检测。
miR-145 在 HCC 组织和 HepG2 细胞中的表达明显低于正常肝组织。转染 miR-145 模拟物后，HepG2 细胞中 miR-145 表达恢复，细胞数量明显减少，G1/S 期转换受到抑制，HepG2 细胞凋亡增加，G1/S 期特异性细胞周期蛋白 E1（CCNE1）和激活的半胱天冬酶-3 水平发生变化。此外，体内外 ROCK1 水平与 miR-145 表达水平相反，共转染 miR-145 模拟物和 pEGFP-N3-3'UTR 的额外实验提供了直接证据表明 ROCK1 基因是 miR-145 的靶基因。此外，ROCK1 表达的显著降低或增加与核因子-kB（NF-κB）（p65）活性有关，脂多糖（LPS）显著增加 NF-κB（p65）活性，同时恢复 miR-145 模拟物对 HepG2 细胞数量的减少。ROCK1 基因在 HepG2 细胞中的过表达显著（p < 0.05，p < 0.01）增加了 NF-κB 活性和细胞数量。
我们表明，miR-145 可以靶向并下调 ROCK1 表达，通过 ROCK1/NF-κB 信号通路抑制细胞周期并激活细胞凋亡来控制 HCC。我们的研究结果为 HCC 的治疗提供了新的视角。
