本研究研究了一种新化合物的合成,PYR26抑制人肝癌HepG2细胞增殖的多靶点机制.PYR26显著抑制HepG2细胞的生长(p<0.0001),并且这种抑制具有浓度效应。PYR26处置后HepG2细胞的ROS释放没有显著变更。CDK4、c-Met和Bak基因在HepG2细胞中的表达受到显著抑制(p<0.05)。而caspase-3、Cytc等促凋亡因子的mRNA表达明显增加(p<0.01)。PI3K的表达,CDK4和pERK蛋白降低。caspase-3蛋白的表达水平升高。PI3K是一种胞内磷脂酰肌醇激酶。PI3K信号通路参与多种生长因子的信号转导,细胞因子和细胞外基质,在防止细胞凋亡中起重要作用,促进细胞存活和影响细胞葡萄糖代谢。CDK4是蛋白激酶复合物的催化亚基,对于细胞周期的G1期进展很重要。PERK是指磷酸化的活化ERK,激活后从细胞质转移到细胞核,然后参与各种生物反应,如细胞增殖和分化,细胞形态维持,细胞骨架构建,细胞凋亡和细胞癌变。与模型组和阳性对照组比较,低浓度PYR26组裸鼠的肿瘤体积,中浓度组和高浓度组较小,脏器体积小于模型组和阳性对照组。PYR26低浓度组、中浓度组、高浓度组的抑瘤率达到50.46%,80.66%和74.59%,分别。结果表明,PYR26通过下调c-Met,抑制HepG2细胞增殖,诱导HepG2细胞凋亡,CDK4和Bak,上调caspase-3和Cytc基因的mRNA表达,下调PI3K,pERK和CDK4蛋白和上调caspase-3的蛋白水平。在一定范围内,随着PYR26浓度的增加,肿瘤生长较慢,肿瘤体积较小。初步结果表明,PYR26对Hepa1-6荷瘤小鼠的肿瘤也有抑制作用。这些结果表明,PYR26对肝癌细胞的生长具有抑制作用,因此,它有潜力发展成为一种新的抗肝癌药物。
This study investigates the synthesis of a new compound, PYR26, and the multi-target mechanism of PYR26 inhibiting the proliferation of HepG2 human hepatocellular carcinoma cells. PYR26 significantly inhibits the growth of HepG2 cells (p < 0.0001) and this inhibition has a concentration effect. There was no significant change in ROS release from HepG2 cells after PYR26 treatment. The mRNA expressions of CDK4, c-Met and Bak genes in HepG2 cells were significantly inhibited (p < 0.05), while mRNA expression of pro-apoptotic factors such as caspase-3 and Cyt c was significantly increased (p < 0.01). The expression of PI3K, CDK4 and pERK proteins decreased. The expression level of caspase-3 protein was increased. PI3K is a kind of intracellular phosphatidylinositol kinase. PI3K signaling pathway is involved in signal transduction of a variety of growth factors, cytokines and extracellular matrix and plays an important role in preventing cell apoptosis, promoting cell survival and influencing cell glucose metabolism. CDK4 is a catalytic subunit of the protein kinase complex and is important for G1 phase progression of the cell cycle. PERK refers to phosphorylated activated ERK, which is translocated from cytoplasm to the nucleus after activation, and then participates in various biological reactions such as cell proliferation and differentiation, cell morphology maintenance, cytoskeleton construction, cell apoptosis and cell canceration. Compared with the model group and the positive control group, the tumor volume of the nude mice in the low-concentration PYR26 group, the medium-concentration group and the high-concentration group was smaller, and the organ volume was smaller than that in the model group and the positive control group. The tumor inhibition rates of low-concentration group PYR26, medium-concentration group and high-concentration group reached 50.46%, 80.66% and 74.59%, respectively. The results showed that PYR26 inhibited the proliferation of HepG2 cells and induced apoptosis of HepG2 cells by down-regulating c-Met, CDK4 and Bak, up-regulating the mRNA expression of caspase-3 and Cyt c genes, down-regulating PI3K, pERK and CDK4 proteins and up-regulating the protein level of caspase-3. In a certain range, with the increase in PYR26 concentration, the tumor growth was slower and the tumor volume was smaller. Preliminary results showed that PYR26 also had an inhibitory effect on the tumors of Hepa1-6 tumor-bearing mice. These results suggest that PYR26 has an inhibitory effect on the growth of liver cancer cells, therefore it has potential to be developed into a new anti-liver cancer drug.