二苯乙烯苷对口腔鳞癌细胞SCC-9增殖、迁移及凋亡的作用机制研究

Effect and mechanism of 2,3,5,4’-tetrahydroxystilbene-2-O-beta-D-glucoside on the proliferation, migration and apoptosis of oral squamous cell carcinoma SCC-9 cells

目的探讨中药单体二苯乙烯苷(TSG)对口腔鳞癌细胞SCC-9增殖、迁移及凋亡的影响及其分子机制。方法将SCC-9细胞分为3组:空白对照组(0μmol·L^(-1)TSG)和低、高2个浓度实验组(14,28μmol·L^(-1)TSG)。用划痕实验和Transwell迁移实验检测细胞迁移水平,用流式细胞仪测定凋亡率,用实时荧光定量-PCR法检测抗凋亡基因B淋巴细胞瘤-2(bcl-2)基因表达水平,用蛋白印迹法检测磷脂酰肌醇3激酶(PI3K)/蛋白激酶B (Akt)/雷帕霉素靶蛋白(mTOR)信号通路相关蛋白表达水平。结果空白对照组和低、高2个浓度实验组于24 h的平均水平迁移率分别为(48.00±5.87)%,(19.30±1.34)%和(17.16±1.50)%;这3组的垂直迁移细胞数分别为73.00±9.17,32.67±3.21和24.33±4.04;这3组的细胞凋亡率分别为(11.10±0.70)%,(23.17±1.03)%和(36.10±1.32)%;这3组的bcl-2基因的相对表达水平分别为1.03±0.02,0.39±0.09和0.15±0.04;这3组的PI3K蛋白表达水平分别为0.18±0.02,0.14±0.01和0.15±0.01;这3组的Akt蛋白表达水平分别为0.26±0.03,0.13±0.01和0.19±0.01;这3组的mTOR蛋白表达水平分别为0.47±0.05,0.38±0.01和0.34±0.02。上述指标:低、高2个浓度实验组与空白对照组相比,差异均有统计学差异(均P<0.05)。结论 TSG具有抑制口腔癌细胞增殖、迁移和促进细胞凋亡的抗癌特性,并可抑制PI3K/Akt/mTOR信号通路相关蛋白表达。

Objective To investigate the effect and mechanism of 2,3,5,4’-tetrahydroxystilbene-2-O-beta-D-glucoside (TSG) on the proliferation,migration and apoptosis of oral squamous carcinoma SCC-9 cells.Methods SCC-9 cells were divided into three groups:Blank control (0μmol·L^(-1)TSG) group,experimental-L,experimental-H(14,28μmol·L^(-1)TSG) groups.The cell migration was detected by scratch assays and Transwell migration assays.The cell apoptosis was determined by flow cytometry.The mRNA expression level of B-cell lymphoma-2 (bcl-2) was detected by quantitative real-time polymerase chain reaction.The protein expression levels of the phosphatidylinositol 3 kinase (PI3 K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway were detected by Western blot.Results The horizontal migration rates at 24 h in blank control group,experimental-L and experimental-H groups were (48.00±5.87)%,(19.30±1.34)%and (17.16±1.50)%,respectively.The number of vertical migration cells in the three groups were 73.00±9.17,32.67±3.21 and 24.33±4.04,respectively.The apoptosis rates in the three groups were(11.10±0.70)%,(23.17±1.03)%and (36.10±1.32)%,respectively.The mRNA expression levels of bcl-2in the three groups were 1.03±0.02,0.39±0.09 and 0.15±0.04,respectively.The protein expression levels of PI3K in the three groups were 0.18±0.02,0.14±0.01 and 0.15±0.01,respectively.The protein expression levels of Akt in the three groups were 0.26±0.03,0.13±0.01 and 0.19±0.01,respectively.The protein expression levels of mTOR in the three groups were 0.47±0.05,0.38±0.01 and 0.34±0.02,respectively.Compared between two experimental groups and blank control group,the difference of the factors were significant (all P<0.05).ConclusionTSG may inhibit the proliferation and migration of SCC-9 cells and promote apoptosis by suppressing the PI3K/Akt/mTOR signaling pathway.