牦牛CCL14蛋白对HepG2细胞活性和凋亡的影响

Effects of Yak CCL14 Protein on Activity and Apoptosis of HepG2 Cells

旨在研究牦牛CCL14蛋白(Bos grunniens C-C motif chemokine 14 protein,BgCCL14)对HepG2细胞的影响和作用机制。将原核表达的BgCCL14蛋白与HepG2细胞共培养,利用CCK-8试剂盒检测细胞活性,平板克隆检测细胞克隆形成能力,细胞划痕检测细胞迁移以及荧光定量PCR检测相关基因的表达量。结果表明,1μg/mL、10μg/mL和20μg/mL的BgCCL14蛋白均能显著降低HepG2细胞活性;20μg/mL的BgCCL14蛋白对细胞增殖和迁移有极显著抑制作用;20μg/mL的BgCCL14蛋白处理36h极显著上调凋亡基因BAK和BAX的mRNA水平,极显著降低HIF1A、PI3K和CDK1基因的mRNA水平,显著降低mTOR基因的mRNA水平。这表明BgCCL14蛋白可能通过促进细胞凋亡抑制肝癌细胞活性。

The aim of this study is to study the effect of Bos grunnies C-C motif chemokine 14 protein(BgCCL14)on HepG2 cells and its mechanism.HepG2 cells were co-cultured with prokaryotic ex­pressed BgCCL14 protein.The cell viability was detected by CCK-8 kit,the ability of cell clone forma­tion was detected by plate clone,the cell migration was detected by scratch test,and the expression of related genes was detected by qPCR.The results showed that BgCCL14 protein of 1 μg/mL,10 μg/mL and 20 μg/mL significantly decreased the viability of HepG2 cells.And BgCCL14 protein of 20 μg/mL significantly inhibited cell proliferation and migration.In addition,BgCCL14 protein of 20 μg/mL extremely significant up-regulate the mRNA levels of apoptotic genes BAK and BAX in 36 h,and decreased mRNA levels of HIF1A,P13K,and CDK1 genes.And the mRNA levels of mTOR al­so significantly decreased.So BgCCL14 may affect viability of HepG2 cells and promotes its apoptosis.This study will benefit for further study of the function of CCL14.