猴头菇多糖联合转化生长因子-β1对胃癌细胞增殖及微小RNA-302a的影响

Hericium erinaceus polysaccharide combined with different doses of TGF-β1 Effects of miR-302a on the prolifteration of gastric cancer ells

目的研究猴头菇多糖联合不同剂量转化生长因子-β1(TGF-β1)对胃癌细胞增殖及微小RNA-302a(miR-302a)的影响。方法培养胃腺癌细胞(低分化:BGC-823、SGC7901);设TGF-β1组、猴头菇多糖+TGF-β1组。并根据TGF-β1的剂量分为0 g/L对照组、1 g/L组、5 g/L组、10 g/L组。噻唑蓝(MTT)法检测两组不同剂量TGF-β1培养6 h、12 h、24 h、48 h和72 h肿瘤细胞的作用后细胞增殖情况;实时荧光定量RT-PCR检测细胞mi R-302a的表达;蛋白印迹法检测磷酸化AKT(p-AKT)、3-羧基磷脂酰肌醇激酶(p-PI3K)蛋白的表达;分别将TGF-β1转染到BGC-823、SGC7901细胞,记录TGF-β1和阴性对照(NC)组。结果两组BGC-823、SGC790细胞随着作用时间的延长和TGF-β1剂量的增大,增殖抑制率也逐渐增加,猴头菇多糖+TGF-β1组增殖抑制率均高于TGF-β1组(P<0.05)。且5、10 g/L TGF-β1组作用48~72 h后对胃癌SGC790细胞的抑制作用相当,较其他作用时间比较均具有统计学意义(P<0.05);与0 g/L比较,两组1、5、10 g/L剂量细胞克隆数量均逐渐降低,10 g/L剂量组细胞克隆数量降低最明显(P<0.05);两组对胃癌细胞BGC-823、SGC7901作用48 h后与0 g/L比较,1、5、10 g/L剂量TGF-β1对胃癌细胞BGC-823、SGC7901凋亡率逐渐增长(P<0.05);猴头菇多糖各剂量组mi R-302a的表达均高于TGF-β1组(P<0.05);各组对p-AKT、p-PI3K的表达随着TGF-β1计量的增加而降低,猴头菇多糖+TGF-β1组降低更显著(P<0.05);蛋白印迹法表明mi R-302a的表达可能受到TGF-β1通过p-AKT、p-PI3K的负调控。结论猴头菇多糖+TGF-β1可更有效地抑制胃腺癌细胞BGC-823、SGC7901的增殖,抑制细胞克隆数量。mi R-302a的表达可能受到TGF-β1通过p-AKT、p-PI3K的负调控。

Objective To study the efcts of hericium ericium polysaccharide combined with different doses of transforming growth factor-B1(TGF-B1)on the proliferation and micro RNA-302a(miR-302a)of gastrie cancer cells.Method Gastrie adenocareinoma cells poorly dferentiated BCC-823 and SGC7901 were culured.TGF-β1 group and hericium erinaceus polysaccharide+TGF-β1 group were set.According to the dose of TGF-β1,0 g/L control group,1 g/L group,5 g/L group and 10 g/L group were divided.MTT assay was used to detect the proliferation of cancer cells after 6 h,12 h,24 h,48 h and 72 h culure of TGF-β1.Real-time fluorescence quantaive RT-PCR was used to detect the expression of miR-302a.The expression of phosphorylated AKT(p-AKT)and 3-carboxphosphatidylinositol kinase(p-PI3K)were detected by Westem boting.TGF-β1 was transfected into BGC-823 and SCC7901 cells,respctively,and TGF-β1 and negative control(NC)groups were recorded.Result The proliferation inhibition rate of BGC--823 and SCC790 cells in two groups increased gradually with the extension of treatment time and the increase of TGF-B1 dose.The proliferation inhibition rate of hericium ericium polysaccharide+TGF-β1 group was higher than that of TGF-β1 group(P<0.05).The inhibitory ffeet of5 and 10 g/L TGF-β1 groups on SGC790 cells was similar afer 48~72 h,which had statistical signifcance compared with other groups(P<0.05).Compared with0 g/L,the number of cell clones in two groups at 1,5 and 10 g/L dose was gradually decreased,and the number of cell clones in 10 g/L dose group was most significantly decreased(P<0.05).Compared with 0 g/L,the apoptosis rate of gastrie cancer cells BGC-823 and SGC7901 was increased by 1,5,10 g/L doses of TGF-B1(P<0.05)after 48 h treatment with TCF-β1(P<0.05).The expression of miR--302a in each dose group was higher than that in TGF-β1 group(P<0.05).The expression of p-Akt and p-PI3K decresed with the increase of dose of TGF-β1 in all groups,and the decrease was more significant in hericium ericium polysaccharide+TGF-B1 group(P<0.05).Westem blotting indicated that the expression of miR-302a may be negaively regulated by TGF-β1 via p-AKT and p-PI3K.Conclusion The proliferation of gastric adenocarcinoma cells BGC-823 and SGC7901 and the number of cell clones were more effectively inhibited by the polyaccharide+TCF-B1.The expression of miR-302a may be negatively regulated by TGF-β1 via p-AKT and p-PI3K.