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 miR-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 miR-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 miR-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。 miR-302a 的表达可能受到 TGF-β1通过 p-AKT、p-PI3K 的负调控。

Objective To study the effects of hericium ericium polysaccharide combined with different doses of transforming growth factor-β1 TGF-β1 on the proliferation and micro RNA-302a miR-302a of gastric cancer cells. Method Gastric adenocarcinoma cells poorly differentiated BGC- 823 and SGC7901 were cultured. 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 culture of TGF - β1. Real-time fluorescence quantitative RT-PCR was used to detect the expression of miR-302a. The expression of phosphorylated AKT p-AKT and 3-carboxyphosphatidylinositol kinase p-PI3K were detected by Western blotting. TGF-β1 was transfected into BGC-823 and SGC7901 cells respectively and TGF-β1 and negative control NC groups were recorded. Result The proliferation inhibition rate of BGC-823 and SGC790 cells in two groups increased gradually with the extension of treatment time and the increase of TGF-β1 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 effect of 5 and 10 g / L TGF-β1 groups on SGC790 cells was similar after 48 ~ 72 h which had statistical significance compared with other groups P < 0. 05 . Compared with 0 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 gastric cancer cells BGC-823 and SGC7901 was increased by 1 5 10 g / L doses of TGF-β1 P< 0. 05 after 48 h treatment with TGF-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 decreased with the increase of dose of TGF-β1 in all groups and the decrease was more significant in hericium ericium polysaccharide +TGF-β1 group P < 0. 05 . Western blotting indicated that the expression of miR- 302a may be negatively 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 polysaccharide +TGF-β1. The expression of miR-302a may be negatively regulated by TGF-β1 via p-AKT and p-PI3K.