Department of Hepatobiliary Surgery, Hanzhong Central Hospital, Hanzhong, Shaanxi 723000, P.R. China.
Mol Med Rep. 2017 Oct;16(4):4348-4354. doi: 10.3892/mmr.2017.7092. Epub 2017 Jul 27.
The abnormal expression of microRNAs (miRNAs) has been reported in hepatocellular carcinoma (HCC), however, the functional role of miR‑143 in HCC remains to be fully elucidated. The present study aimed to investigate the effects of the downregulation of miR‑143 on HCC cell proliferation and apoptosis, and elucidated the underlying mechanism. Hepg2 and Hep3B human hepatoma cell lines cells were transfected with miR‑143 inhibitor. Following transfection, the cell viability and apoptosis were respectively determined using a 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2‑H‑tetrazolium bromide assay and flow cytometry, and the mRNA and protein levels of protein kinase Cε (PKCε) were examined. The expression levels of PKCε were downregulated by short hairpin (sh)RNA, and the effects of the downregulation of miR‑143 on HCC cell proliferation were measured. The results showed that the miR‑143 inhibitor significantly promoted cell proliferation and suppressed apoptosis in the Hepg2 and Hep3B cells. The miR‑143 inhibitor significantly increased the protein levels of PKCε in the Hepg2 and Hep3B cells; however, no significant differences were found in the mRNA levels of PKCε. In addition, the downregulation of PKCε markedly decreased the cell viability of the Hepg2 and Hep3B cells, and co‑transfection with the miR‑143 inhibitor and PKCε shRNA significantly alleviated the miR‑143 inhibitor‑induced high cell proliferation. Taken together, these results suggested that miR‑143 acts as a tumor suppressor gene in HCC. The downregulation of mi‑143 promoted cell proliferation by regulating PKCε in the HCC cells.
miRNAs(miRs)的异常表达已在肝细胞癌(HCC)中报道,然而,miR-143 在 HCC 中的功能作用仍有待充分阐明。本研究旨在探讨下调 miR-143 对 HCC 细胞增殖和凋亡的影响,并阐明其潜在机制。将 miR-143 抑制剂转染 Hepg2 和 Hep3B 人肝癌细胞系。转染后,分别采用 3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四唑溴盐法和流式细胞术检测细胞活力和凋亡,检测蛋白激酶 Cε(PKCε)的 mRNA 和蛋白水平。通过短发夹(sh)RNA 下调 PKCε 的表达,并测定下调 miR-143 对 HCC 细胞增殖的影响。结果表明,miR-143 抑制剂显著促进 Hepg2 和 Hep3B 细胞的增殖并抑制凋亡。miR-143 抑制剂显著增加 Hepg2 和 Hep3B 细胞中 PKCε 的蛋白水平;然而,PKCε 的 mRNA 水平没有明显差异。此外,下调 PKCε 显著降低 Hepg2 和 Hep3B 细胞的活力,并且 miR-143 抑制剂和 PKCε shRNA 的共转染显著减轻了 miR-143 抑制剂诱导的高细胞增殖。总之,这些结果表明 miR-143 在 HCC 中作为肿瘤抑制基因发挥作用。miR-143 通过调节 HCC 细胞中的 PKCε 促进细胞增殖。