Objective: To study the inhibitory effect of 10-gingerol on the proliferation of hepatocellular carcinoma HepG2 cells and the role of Src/STAT3 signaling pathway in mediating the effect.
Methods: SYBYL-X2.1 software was used to simulate the interaction between 10-gingerol and Src. HepG2 cells treated with 10-gingerol at 1, 3, 10 or μol/L for 24 h were assessed for cell viability using MTT assay, and EdU staining was used to detect the cell proliferation and calculate the number of positive cells. The expressions of p-Src and p-STAT3 were detected using Western blotting, and the mRNA expressions of the target genes of STAT3 (cyclin D1 and CMCC) were detected using qPCR.
Results: 10-gingerol was capable of forming hydrogen bond with such Src residues as TRY-340, MET-341, MET-314, ASP-404, and ILE-336. MTT assay showed that 10-gingerol at 3 and 10 μmol/L significantly lowered the viability of HepG2 cells (P < 0.001). Treatment with 1, 3, and 10 μmol/L 10-gingerol significantly reduces the number of EdU-positive HepG 2 cells (P < 0.001). Western blotting showed that 10-gingerol at 3 and 10 μmol/L significantly decreased the phosphorylation levels of Src and STAT3 in HepG2 cells (P < 0.01). 10-gingerol at 1, 3, and 10 μmol/L significantly decreased the mRNA expressions of cyclin D1 and CMCC as shown by qPCR (P < 0.01).
Conclusions: 10-gingerol can dose-dependently inhibit the proliferation of HepG2 cells and suppress the activation of Src and STAT3.
目的: 研究10-姜酚通过Src/STAT3信号通路抑制肝癌HepG2细胞增殖。
方法: 使用SYBYL-X2.1软件对10-姜酚与Src之间的相互结合进行模拟。选择10-姜酚低、中、高浓度(1、3、10 μmol/L)处理肝癌细胞HepG2 24 h，同时设立正常对照组。MTT检测各组细胞活力；EdU染色检测各组细胞增殖并计算阳性细胞数；Western blot检测Src(p-Src)及STAT3(p-STAT3)的表达水平；qPCR技术检测STAT3靶基因CyclinD1和cmyc的表达。
结果: 10-姜酚可以与Src的氨基酸残基TRY-340、MET-341、MET-314、ASP404、ILE-336形成氢键连接。与对照组相比，MTT显示，10-姜酚(3、10 μmol/L)显著降低HepG2的细胞活性(P < 0.001)；EdU染色显示，10-姜酚(1、3、10 μmol/L)均能显著降低HepG2细胞EdU染色阳性细胞数(P < 0.001)；Western blot显示，10-姜酚(3、10 μmol/L)显著降低HepG2的细胞Src及STAT3的磷酸化表达水平(P < 0.01)；qPCR显示，10-姜酚(1、3、10 μmol/L)显著降低STAT3靶基因CyclinD1和cmyc的基因表达水平(P < 0.01)。
结论: 10-姜酚能剂量依赖性的抑制肝癌HepG2细胞的增殖，抑制Src及其下游的STAT3的激活。
Keywords: 10-gingerol; Src/STAT3; hepatocellular carcinoma.