Department of Gastroenterology, The First Peoples' Hospital of Tianmen City, Tianmen, Hubei, China (mainland).
Med Sci Monit. 2019 Aug 13;25:6023-6033. doi: 10.12659/MSM.914768.
BACKGROUND The PCK1 gene encodes phosphoenolpyruvate carboxykinase (PEPCK), which has been shown have a role in metabolic events in hepatocellular carcinoma (HCC). This study aimed to investigate the role of the SHH gene and its encoded protein, sonic hedgehog (SHH), in two human hepatocellular carcinoma (HCC) cell lines. MATERIAL AND METHODS The human HCC cell lines Hep3B and SMMC-7721 were cultured. Cells were transfected with plasmids carrying specific SHH gene short-hairpin RNA (shRNA) and negative control (NC) shRNA. The effects of knockdown of expression levels of the SHH gene were studied on cell survival, cell apoptosis, the cell cycle, gluconeogenesis, and the expression of PCK1. Anchorage-independent growth, a characteristic of transformed cells, was detected by the colony formation assay. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western blot were performed 24 h after transfection. RESULTS Knockdown of expression levels of the SHH gene reduced cell proliferation and growth of HCC cells and induced cell apoptosis and G1 cell cycle arrest in both HCC cell lines. Knockdown of the SHH gene decreased the levels of glycolysis products and increased the production of glucose and reduced the phosphorylation of PI3K and Akt but induced the expression of PCK1. CONCLUSIONS Knockdown of the SHH gene reduced cell survival of HCC cells by increasing apoptosis, reducing cell proliferation, inducing G1 cell cycle arrest, and restoring gluconeogenesis, and was associated with the inhibition of the PI3K/Akt axis and induced the expression of PCK1.
PCK1 基因编码磷酸烯醇丙酮酸羧激酶（PEPCK），它在肝细胞癌（HCC）的代谢事件中发挥作用。本研究旨在研究 SHH 基因及其编码蛋白 sonic hedgehog（SHH）在两种人肝癌（HCC）细胞系中的作用。
培养人 HCC 细胞系 Hep3B 和 SMMC-7721。用携带特定 SHH 基因短发夹 RNA（shRNA）和阴性对照（NC）shRNA 的质粒转染细胞。研究了 SHH 基因表达水平降低对细胞存活、细胞凋亡、细胞周期、糖异生和 PCK1 表达的影响。通过集落形成试验检测转化细胞的特征性无锚定生长。转染 24 h 后进行逆转录定量聚合酶链反应（RT-qPCR）和 Western blot。
SHH 基因表达水平的降低降低了两种 HCC 细胞系中 HCC 细胞的增殖和生长，并诱导细胞凋亡和 G1 细胞周期阻滞。SHH 基因的敲低降低了糖酵解产物的水平，增加了葡萄糖的产生，降低了 PI3K 和 Akt 的磷酸化，但诱导了 PCK1 的表达。
通过增加细胞凋亡、降低细胞增殖、诱导 G1 细胞周期阻滞和恢复糖异生，降低 SHH 基因的表达降低了 HCC 细胞的存活率，与抑制 PI3K/Akt 轴和诱导 PCK1 表达有关。
