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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 detect the effect of yak hepatic protein BGP on the proliferation and apoptosis of HepG2 human hepatoma cells and to investigate its antitumor activity in vitro. Methods HepG2 cells were treated with different concentrations of BGP. The cells were treated with BGP (25, 50 and 100 mg / L) for 12, 24, 36 and 48 h on the proliferation of cells, and observed under inverted microscope cell morphology; using flow cytometry BGP (25,50,75,100 mg / L) HepG2 cells for 24 h, the cells Cycle and apoptosis. Results The proliferation of HepG2 cells was inhibited by different concentrations of BGP. Compared with untreated cells, the difference was statistically significant (P <0.05). Among them, BGP treatment with 100 mg / L of BGP for 48 h resulted in the highest inhibitory rate of HepG2 cells 94.99%). HepG2 cells in each concentration of BGP group changed significantly, the interstitial space increased, the intercellular connection was not firm, intracellular particles increased, showing the growth retardation state, even cells shedding, the number of cells decreased, of which 100 mg / L BGP effect 48 h, apoptotic cells increased significantly. After treated with different concentrations of BGP for 24 h, HepG2 cells could inhibit HepG2 cell growth and induce apoptosis in different degrees. Compared with untreated cells, the proportion of S phase cells was significantly increased (except for 25 mg / L BGP group) , The proportion of cells in G0 / G1 phase and G2 / M phase decreased significantly, the differences were statistically significant (P <0.05). Conclusion Yak hepatic protein BGP can inhibit HepG2 cell proliferation, promote apoptosis, and have obvious anti-hepatoma activity in vitro.