牦牛肝脏蛋白对人肝癌HepG2细胞增殖及凋亡的影响

Effect of yak liver protein on proliferation and apoptosis of human hepatocarcinoma HepG2 cells

目的检测牦牛肝蛋白BGP对人肝癌HepG2细胞增殖及凋亡的影响,并探讨其体外抗癌活性。方法用不同浓度的BGP分别作用于人肝癌HepG2细胞,作为实验组,同时设未处理细胞对照组和空白对照组,采用MTT法检测BGP(25、50和100 mg/L)作用HepG2细胞12、24、36和48 h对细胞增殖活性的影响,并于倒置显微镜下观察细胞形态变化;采用流式细胞仪检测BGP(25、50、75、100 mg/L)作用HepG2细胞24 h,对细胞周期及凋亡的影响。结果不同浓度的BGP均可抑制HepG2细胞增殖,与未处理细胞对照组相比,差异均有统计学意义(P<0.05),其中100 mg/L BGP作用48 h,对HepG2细胞抑制率最高(94.99%)。各浓度BGP实验组HepG2细胞形态发生明显改变,细胞间隙加大,细胞间连接不牢固,胞内颗粒增多,呈现生长受阻状态,甚至有细胞脱落,细胞数目减少,其中100 mg/L BGP作用48 h,凋亡细胞明显增多。不同浓度BGP作用HepG2细胞24 h后,均可不同程度抑制HepG2细胞生长并诱导细胞凋亡;与未处理细胞对照组相比,S期细胞比例明显升高(除25 mg/L BGP实验组),G0/G1期和G2/M期细胞比例明显下降,差异均有统计学意义(P<0.05)。结论牦牛肝蛋白BGP可抑制HepG2细胞增殖,并促进凋亡,具有明显的体外抗肝癌活性。

Objective To investigate the in vitro antitumor activity of yak （Bos grunnisens ） liver protein BGP as well as its effect on proliferation and apoptosis of human hepatocarcinoma HepG2 cells. Methods HepG2 cells were treated with BGP at various concentrations, using those untreated as negative control. Blank control was set up. The proliferative activities of HepG2 cells 12, 24, 36 and 48 h after treatment with 25, 50 and 100 mg/L BGP were determined by MTT assay, while the morphologies were observed under inverted microscope. The cell cycle and apoptosis of HepG2 ceils 24 h after treatment with 25, 50, 75 and 100 mg/L BGP were determined by flow cytometry. Results The proliferations of HepG2 cells treated with BGP at various concentrations were inhibited significantly as compared with those untreated（P 〈 0. 05）, of which the inhibition rate reached the maximum（94. 99%） 48 h after treatment with 100 mg/L BGP. The morphologies of HepG2 cells in various test groups changed significantly, expressed as increased space and loose junction between cells, increased intracellular particles, arrested growth, even death, and decreased count of cells. The count of apoptotic cells increased significantly 48 h after treatment with 100 mg/L BGP. The treatment with BGP at various con- centrations for 24 h inhibited the growth and induced the apoptosis of HepG2 cells at different degrees. The percentage of cells after treatment with BGP（except that at a concentration of 25 mg/L） at S phase increased significantly, while that at GO/G1 and G2/M phases decreased significantly （each P 〈 0. 05）. Conclusion BGP inhibited the proliferation and promoted the apoptosis of HepG2 cells, which showed significantly antitumor activity in vitro.