Medical College of Nanchang University, Nanchang 330006, China.
Department of Elderly Oncology, Jiangxi Provincial Tumor Hospital, Nanchang 330029, China.
J Nanosci Nanotechnol. 2019 Sep 1;19(9):5476-5482. doi: 10.1166/jnn.2019.16535.
MicroRNAs play an important role in cell proliferation and migration in hepatocellular carcinoma (HCC). In this study, the expression of in the liver cancer tissues of 116 patients, and in one normal (HL-7702) and five HCC cell lines (HepG2, SMMC-7721, Bel-7402, QGY-7701 and QGY-7703) was determined by qRT-PCR. Results indicated that was significantly repressed in HCC patients and in HCC cell lines, especially in the Bel-7402 cells when compared to normal liver tissue and the normal hepatocyte line, HL-7702. Therefore, artificial mimic computationally targeting E2F1 and anti-sense inhibitor were designed and used to transfect the Bel-7402 lines. Stable expression and significant inhibition of , with clear changes in cell morphology, proliferation and motility, were detected in Bel-7402 cells transfected with mimic or inhibitor, respectively. As E2F1 was computationally recognized as the target of , the expression of E2F1 in transfected Bel-7402 cells was analyzed by qRT-PCR and Western blotting. Results showed a strong inhibition of E2F1 by and further promotion of E2F1 by inhibitor in transfected Bel-7402 cells. Flow cytometry analysis showed that the cell ratio was restored to S to G0/1 cell number. Inhibition of E2F1 by mimic promoted cell proliferation and migration in Bel-7402. These results demonstrated that down-regulated E2F1 to promote cell proliferation and migration in HCC.
微小 RNA 在肝癌（HCC）的细胞增殖和迁移中发挥重要作用。在这项研究中，通过 qRT-PCR 检测了 116 例肝癌组织、1 个正常（HL-7702）和 5 个 HCC 细胞系（HepG2、SMMC-7721、Bel-7402、QGY-7701 和 QGY-7703）中 的表达。结果表明，与正常肝组织和正常肝细胞系 HL-7702 相比，HCC 患者和 HCC 细胞系中 显著下调，尤其是 Bel-7402 细胞。因此，设计并使用人工 miRNA 模拟物靶向 E2F1 和反义 miRNA 抑制剂转染 Bel-7402 细胞系。在转染 miRNA 模拟物或抑制剂的 Bel-7402 细胞中，检测到 的稳定表达和明显抑制，细胞形态、增殖和迁移均发生明显变化。由于 E2F1 被计算为 的靶标，通过 qRT-PCR 和 Western blot 分析转染的 Bel-7402 细胞中 E2F1 的表达。结果显示，miRNA 模拟物强烈抑制 E2F1，并进一步促进转染的 Bel-7402 细胞中 E2F1 抑制剂的表达。流式细胞术分析表明，细胞比例恢复到 S 期到 G0/1 期细胞数。miRNA 模拟物抑制 E2F1 促进 Bel-7402 细胞增殖和迁移。这些结果表明，下调 E2F1 促进 HCC 中的细胞增殖和迁移。
