白花蛇舌草总黄酮对人结直肠癌细胞增殖和凋亡的影响  

Effects of total flavonoids of Baihua Sheshecao on proliferation and apoptosis of human colorectal cancer cells

作　　者：周剑飞 陈思瑾 ZHOU Jianfei

机构地区：[1]广西玉林市第二人民医院,广西玉林537000

出　　处：《中医临床研究》2024年第19期49-54,共6页Clinical Journal Of Chinese Medicine

摘　　要：目的:探究白花蛇舌草总黄酮对人结直肠癌细胞增殖和凋亡的影响。方法:体外培养HCT116细胞,用不同浓度白花蛇舌草总黄酮进行处理,分组:0μg/mL组、1μg/mL组、3μg/mL组、6μg/mL组、12μg/mL组。采用细胞计数试剂盒法检测24 h和48 h各组HCT116细胞的增殖能力,并计算抑制率。用苏木精-伊红染色观察细胞形态,用流式细胞术检测HCT116细胞凋亡情况,用实时荧光定量逆转录聚合酶链反应(Real-Time Fluorescence PCR,qRT-PCR)检测B淋巴细胞瘤-2(B-cell Lymphoma-2,Bcl-2)、Bcl2关联X蛋白(Bcl2 Associated X,Bax)和半胱氨酸蛋白酶3(Caspase-3)的信使核糖核酸(Messenger RNA,mRNA)表达水平,用蛋白质印迹(Western blot)法检测Bcl-2、Bax、Caspase-3和裂解的Caspase-3的蛋白表达水平。结果:不同浓度的白花蛇舌草总黄酮能明显抑制癌细胞增殖,其浓度越高,抑制率越高(均P <0.05),且处理组48 h抑制率均高于24 h(均P <0.05)。随着浓度的增加,细胞核逐渐收缩,核碎片增加,同时凋亡小体产生。与对照组相比,白花蛇舌草总黄酮也显著促进HCT116细胞的凋亡率(P <0.05)。此外,与对照组相比,白花蛇舌草总黄酮显著下调了Bcl-2和Caspase-3的表达,同时上调Bax和裂解的Caspase-3的表达,并且Bcl-2/Bax值显著降低,而裂解的Caspase-3/Caspase-3值显著升高。结论:白花蛇舌草总黄酮可抑制结直肠癌细胞的体外增殖,并通过影响Bcl-2、Bax和Caspase-3的表达诱导细胞凋亡。Objective:To investigate the effects of total flavonoids of Oldenlandia diffusa on the proliferation and apoptosis of Ohuman colorectal cancer cells.Methods:HCT116 cells were cultured in vitro and treated with different concentrations of total flavonoids of Oldenlandia diffusa in groups of 0μg/mL,1μg/mL,3μg/mL,6μg/mL,and 12μg/mL.CCK-8 assay was used to detect the proliferation ability of HCT116 cells in each group at 24 h and 48 h,and the inhibition rate was calculated.H&E staining was used to observe the cell morphology.The apoptosis of HCT116 cells was detected by flow cytometry,the mRNA expression levels of Bcl-2,Bax and Caspase 3 were detected by qRT-PCR,and the protein expression levels of Bcl-2,Bax,Caspase-3 and Cleaved-Caspase-3 were detected by Western blot.Results:Different concentrations of total flavonoids of Oldenlandia diffusa could significantly inhibit the proliferation of cancer cells,and the higher the concentration,the higher the inhibition rate(all P<0.05),and the inhibition rate in the treated group was higher at 48 h than that at 24 h(all P<0.05).As the concentration increased,the nucleus gradually contracted and nuclear fragmentation increased,while apoptotic vesicles were produced.Total flavonoids of Oldenlandia diffusa also significantly promoted the apoptosis rate of HCT116 cells compared with the control group(P<0.05).In addition,compared with the control group,total flavonoids of Oldenlandia diffusa significantly down-regulated the expression of Bcl-2 and Caspase-3,while up-regulated the expression of Bax and Cleaved Caspase-3,and the Bcl-2/Bax value was significantly lower,while the Cleaved Caspase-3/Caspase-3 value was significantly higher.Conclusion:Total flavonoids of Oldenlandia diffusa inhibits the proliferation of colon cancer cells in vitro and induce apoptosis by affecting the expressions of Bcl-2,Bax and Caspase-3.

关 键 词：白花蛇舌草总黄酮 结直肠癌 增殖 凋亡 

分 类 号：R735.37[医药卫生—肿瘤]