miR-106a通过调控PI3K/PDK1/AKT蛋白通路调节胃癌细胞生物学行为的研究

miR-106a regulates the biological behavior of gastric cancer cells through the regulation of PI3K/PDK1/AKT pathway

目的探讨miR-106a对胃癌细胞生物学行为的影响及其作用机制。方法选取AGS人胃癌细胞系,经培养后分为27个样本。所有样本随机分为miR-106a inhibitor、miR-mimic、miR-NC共计3组,分别给予miR-106a抑制剂、miR-106a模拟物及安慰剂干预。观察各组细胞存活率、细胞周期、细胞侵袭、迁移及caspase活性、Bax、Bcl-2、Casepase-3蛋白相对表达量、p85β、p-PDK1、p-AKT蛋白相对表达量。结果与miR-NC组比较,miR-106a inhibitor组AGS细胞活性降低[(分别为15.01±0.97)、(69.82±2.31)%](P<0.01);miR-106a mimics组AGS细胞G0/G1期细胞比例降低(P<0.05)(分别为17.33±1.04、58.24±0.82),G2/M和S期细胞比例升高(分别为50.11±1.12、35.64±1.07和31.56±0.92、9.24±0.25);miR-106a inhibitor组AGS细胞G0/G1期细胞比例升高(分别为78.43±1.12、58.24±0.82)(P<0.05),G2/M和S期细胞比例降低(分别为33.65±0.99、35.64±1.07和19.78±0.84、9.24±0.25)(P<0.01)。与miR-NC相比,miR-106a mimics组AGS细胞的迁移、侵袭能力增强,miR-106a inhibitor组AGS细胞的迁移、侵袭能力减弱(P<0.01);与miR-NC相比,miR-106a mimics组AGS细胞caspase-3、caspase-8、caspase-9活性降低,miR-106a inhibitor组AGS细胞caspase-3、caspase-8、caspase-9活性升高(P<0.05);miR-NC相比,miR-106a mimics组AGS细胞Bax(分别为0.69±0.07、1.48±0.15)、Casepase-3蛋白(分别为0.37±0.04、0.91±0.09)相对表达量降低,Bcl-2蛋白相对表达量升高(分别为1.53±0.12、0.94±0.09),miR-106a inhibitor组AGS细胞Bax(分别为2.07±0.21、1.48±0.15)、Casepase-3蛋白(分别为1.23±0.12、0.91±0.09)相对表达量升高,Bcl-2蛋白相对表达量降低(P<0.05)(分别为0.65±0.07、0.94±0.09);与miR-NC相比,miR-106a mimics组AGS细胞p85β(分别为1.24±0.12、0.94±0.09)、p-PDK1(分别为2.13±0.23、1.01±0.10)、p-AKT蛋白(分别为1.14±0.11、0.72±0.06)相对表达量升高,miR-106a inhibitor组AGS细胞p85β(分别为0.69±0.07、0.94±0.09)、p-PDK1(分别为0.75±0.07、1.01±0.10)、p-AKT(分别为0.53±0.05、0.72±0.06)相对表达量降低(P<0.05)。结论miRNA-106a表达能通过调控宫颈癌细胞PI3K/PDK1/AKT蛋白通路调控其细胞生物学行为,包括降低癌细胞活力、迁移和侵袭,诱导癌细胞细胞周期停滞,抑制miRNA-106a表达可能是胃癌患者治疗的新靶点之一。

Objective To investigate the effects of miR-106a on the biological behavior of gastric cancer cells and its mechanism.Methods Human gastric cancer cell lines of AGS were selected and cultured into 27 sam-ples.All the samples were randomly divided into three groups:miR-106a inhibitor,miR-mimic and miR-NC,which were treated with miR-106a inhibitor,miR-106a mimic and placebo respectively.Cell survival rate,cell cycle,cell invasion,migration,caspase activity,relative expression levels of Bax,Bcl-2,Casepase-3 protein,and relative ex-pression levels of p85β,p-PDK1 and p-AKT protein were observed in each group.Results Compared with miR-NC,AGS cell activity of miR-106a inhibitor group was decreased(15.01±0.97 vs 69.82±2.31)(P<0.01).The pro-portion of G0/G1 phase cells in AGS cells of miR-106a mimics group decreased(P<0.05)(17.33±1.04 vs 58.24±0.82).The proportion of G2/M and S phase cells increased(50.11±1.12 vs 35.64±1.07)(31.56±0.92 vs 9.24±0.25).The proportion of AGS cells in G0/G1 phase was increased(78.43±1.12 vs 58.24±0.82)(P<0.05)after miR-106a in-hibitor treatment.The proportion of G2/M and S phase cells decreased(33.65±0.99 vs 35.64±1.07)(19.78±0.84 vs 9.24±0.25)(P<0.01);Compared with miR-NC,the migration and invasion ability of AGS cells in miR-106a mimics group was enhanced.The migration and invasion ability of AGS cells were decreased vs miR-106a inhibitor group(P<0.01).Compared with miR-NC,the activities of caspase-3,caspase-8 and caspase-9 of AGS cells in miR-106a mimics group were decreased.The activities of caspase-3,caspase-8 and caspase-9 of AGS cells in miR-106a inhibi-tor group were increased(P<0.05).Compared with miR-NC,the relative expression levels of Bax and Casepase-3 in AGS cells of miR-106a mimics group were decreased(0.69±0.07 vs 1.48±0.15)(0.37±0.04 vs 0.91±0.09).The rela-tive expression of Bcl-2 protein was increased(1.53±0.12 vs 0.94±0.09).The relative expression levels of Bax and Casepase-3 in AGS cells of miR-106a inhibitor group were increased(2.07±0.21 vs 1.48±0.15)(1.23±0.12 vs 0.91±0.09).The relative expression of Bcl-2 protein was decreased(P<0.05)(0.65±0.07 vs 0.94±0.09).Compared with miR-NC.The relative expression levels of p85β,p-PDK1 and p-AKT in AGS cells of miR-106a mimics group were increased(1.24±0.12 vs 0.94±0.09)(2.13±0.23 vs 1.01±0.10)(1.14±0.11 vs 0.72±0.06).The relative expression levels of p85β,p-PDK1 and P-Akt in AGS cells of miR-106a inhibitor group were decreased(0.69±0.07 vs 0.94±0.09)(0.75±0.07 vs 1.01±0.10)(0.53±0.05 vs 0.72±0.06)(P<0.05).Conclusion The expression of miRNA-106a can regulate the biological behavior of cervical cancer cells through the regulation of PI3K/PDK1/AKT path-way,including reducing the viability,migration and invasion of cancer cells,and inducing cell cycle arrest of can-cer cells.Inhibiting the expression of miRNA-106a may be a new therapeutic target for patients with gastric cancer.