蜂王浆蛋白体外消化产物对胃癌细胞SGC-7901增殖和凋亡的影响及其可能机制

Effects of royal jelly protein in vitro digestion products on proliferation and apoptosis of gastric cancer cell SGC-7901 and its possible mechanism

通过研究蜂王浆蛋白体外消化产物对胃癌细胞SGC-7901增殖、凋亡的影响,并探讨其可能的作用机制,为蜂王浆蛋白辅助治疗胃癌奠定实验基础。采用碱提酸沉法提取蜂王浆蛋白,并通过体外模拟消化实验得到蜂王浆蛋白体外消化产物,然后用不同质量浓度的蜂王浆蛋白体外消化物诱导体外培养的胃癌细胞SGC-7901,通过倒置显微镜观察其对胃癌细胞形态的影响,并通过集落形成实验检测诱导后胃癌细胞的集落形成能力;采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)法检测其对胃癌细胞增殖的抑制作用,通过流式细胞术检测细胞周期及凋亡,并通过蛋白质印迹法检测其诱导后胃癌细胞中p53和PARP-1蛋白的表达情况。结果表明:蜂王浆蛋白体外消化产物使胃癌细胞形态发生皱缩,细胞密度降低,集落形成能力降低(P<0.05),对胃癌细胞SGC-7901的增殖具有抑制作用。0.2 mg/mL体外消化产物经24 h和48 h培养后对胃癌细胞抑制率分别达到(57.58±3.48)%和(62.84±1.98)%(P<0.05)。与空白对照组相比,蜂王浆蛋白体外消化产物对SGC-7901细胞增殖的抑制作用呈剂量依赖性,S期细胞减少,从而引起细胞凋亡;且经蜂王浆蛋白体外消化产物诱导后,胃癌细胞中p53蛋白表达量增加(P<0.05),PARP-1蛋白表达量减少(P<0.05)。说明蜂王浆蛋白体外消化产物能抑制胃癌细胞SGC-7901的增殖并诱导其发生凋亡,其机制可能与p53表达上调和PARP-1表达下调有关。

To study the effects of in vitro digestion products of royal jelly protein on the proliferation and apoptosis of gastric cancer cell SGC-7901 and to explore its possible mechanism so as to provide an experimental basis for the royal jelly protein in the treatment of gastric cancer,the royal jelly protein was extracted by alkali extraction and acid precipitation.The in vitro digestion products of royal jelly protein were obtained by in vitro simulated digestion experiments.The gastric cancer cells SGC-7901 were cultured in vitro with different concentrations of royal jelly protein in vitro digestion products,and the influence on gastric cancer cells were observed by an inverted microscope.The colony formation ability of gastric cancer cells after induction was tested.The 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide(MTT)assay was used to detect the inhibition against proliferation of the gastric cancer cells;the cell cycle and apoptosis were detected by flow cytometry;and the expression levels of p53 and PARP-1 proteins in the gastric cancer cells were detected by Western blotting.The results showed that the in vitro digestion products of royal jelly protein leaded to the shrinkage of gastric cancer cells,decreased cell density and colony formation ability(P<0.05),and inhibited the proliferation of gastric cancer cell SGC-7901.When the in vitro digestion product concentration was 0.2 mg/mL,the inhibition rates at 24 h and 48 h were(57.58±3.48)%and(62.84±1.98)%,respectively(P<0.05).Compared with the blank control group,the in vitro digestion products of royal jelly protein inhibited the proliferation of SGC-7901 cells in a dose-dependent manner,and the S phase cells decreased,which leading to the apoptosis.After induction by in vitro digestion products of royal jelly protein,the expression of p53 increased in the gastric cancer cells(P<0.05),and the expression level of PARP-1 protein decreased(P<0.05).The above results show that the in vitro digestion products of royal jelly protein can inhibit the proliferation of gastric cancer cell SGC-7901 and induce its apoptosis.The mechanism may be related to the up-regulation of p53 expression and the down-regulation of PARP-1 expression.