重楼皂苷调控PI3K/AKT/m TOR通路抑制皮肤基底细胞癌细胞迁移和侵袭

Dioscin inhibits the migration and invasion of cutaneous basal cell carcinoma cells by regulating the PI3K/AKT/m TOR target of rapamycin pathway

目的探讨重楼皂苷对皮肤基底细胞癌细胞迁移、侵袭的影响及其相关机制。方法取对数生长期皮肤基底细胞癌TE 354.T细胞,将其随机分为对照组、低剂量实验组、中剂量实验组、高剂量实验组。低、中、高剂量实验组分别以浓度为20,40,60μg·mL^(-1)的重楼皂苷处理,对照组使用等量二甲基亚砜处理,各组均干预24 h。用细胞计数试剂盒-8实验检测细胞增殖能力,用流式细胞术检测细胞凋亡能力,用划痕实验来检测细胞迁移能力,用Transwell法检测细胞的侵袭能力,用蛋白印迹法检测细胞中磷酸化磷脂酰肌醇3-激酶(p-PI3K)、磷酸化蛋白激酶B(p-Akt)和磷酸化哺乳动物雷帕霉素靶蛋白(p-mTOR)的表达。结果对照组、低、中、高剂量实验组细胞存活率分别为(100.00±0.00)%,(85.86±8.93)%,(67.24±9.21)%和(53.19±8.06)%;这4组的细胞凋亡率分别为(3.02±0.58)%,(8.15±1.23)%,(16.92±2.16)%和(33.70±2.25)%;这4组的迁移率分别为(79.52±5.91)%,(59.68±5.26)%,(32.09±4.12)%和(21.33±3.62)%;这4组的细胞侵袭数分别为94.52±9.23,72.41±6.01, 61.24±5.49和44.35±5.46;这4组的p-PI3K蛋白表达量分别为1.25±0.26,0.98±0.14,0.67±0.11和0.41±0.09。低、中、高剂量实验组与对照组比较,差异均有统计学意义(均P<0.05)。结论重楼皂苷可显著抑制皮肤基底细胞癌细胞迁移和侵袭,其作用机制可能与下调p-Akt、p-mTOR和p-PI3K蛋白表达有关。

Objective To investigate the effect of dioscin on the migration and invasion of cutaneous basal cancer cells and its mechanism.Methods TE 354.T cells of logarithmic growth skin basal cell carcinoma were randomly divided into control group,experimental-L group,experimental-M group and experimental-H group.The experimental-L group,experimental-M group and experimental-H group were treated with 20,40 and 60μg·m L^(-1)dioscin,respectively,and the control group was treated with 150μL dimethyl sulfoxide for24 h.Cell counting kit-8 assay was used to detect cell proliferation,flow cytometry was used to detect cell apoptosis,scratch assay was used to detect cell migration,Transwell assay was used to detect cell invasion.Western blotting was used to detect phospho-phosphoinositide-3 kinase(P-PI3K),phospho-protein kinase B (P-akt) and phosphomammalian target of rapamycin (P-MTOR).Results The cell survival rate of control group,experimental-L group,experimental-M group and experimental-H group were (100.00±0.00)%,(85.86±8.93)%,(67.24±9.21)%and (53.19±8.06)%,respectively;the apoptosis rate of these 4 groups were (3.02±0.58)%.(8.15±1.23)%,(16.92±2.16)%and (33.70±2.25)%;the mobility of the four groups were(79.52±5.91)%,(59.68±5.26)%,(32.09±4.12)%and (21.33±3.62)%,respectively;the number of cell invasion were 94.52±9.23,72.41±6.01,61.24±5.49 and 44.35±5.46,respectively;the P-pi3k protein expression were 1.25±0.26,0.98±0.14,0.67±0.11 and 0.41±0.09,respectively.The experimental-L,-M,-H group compared with the control group,the differences were statistically significant (all P<0.05).ConclusionDioscin can significantly inhibit the migration and invasion of cutaneous basal cancer cells,the mechanism of which may be related to the down-regulation of p-Akt,p-m TOR and p-PI3K protein expressions.