半枝莲总黄酮调控MIIP对人胃腺癌细胞AGS增殖、凋亡和放疗敏感性的作用及机制研究  

Effects of total flavonoids of Scutellaria barbata on proliferation, apoptosis and radiotherapy sensitivity of gastric cancer cell AGS by regulating MIIP

作　　者：张锋[1] 吴震[1] 徐宁 杜红蕾 郭海彦 ZHANG Feng;WU Zhen;XU Ning;DU Honglei;GUO Haiyan(The second Central Hospital of Baoding,Baoding 071000,China)

机构地区：[1]河北省保定市第二中心医院,河北保定071000

出　　处：《药物评价研究》2022年第4期686-692,共7页Drug Evaluation Research

摘　　要：目的研究半枝莲总黄酮(TF-SB)对人胃腺癌细胞AGS增殖、凋亡和放疗敏感性的影响和分子机制.方法采用细胞计数试剂盒(CCK-8)法检测TF-SB(0、25、50、100、200μg·mL^(-1))作用24 h对AGS细胞存活率的影响.将AGS细胞分为对照组、溶剂对照(含相同浓度的甲醇)组、TF-SB(100μg·mL^(-1))组、质粒空载体(pcDNA,200nmol·L^(-1))组、迁移侵袭抑制蛋白(MIIP)过表达质粒(pcDNA-MIIP,200nmol·L^(-1))组、TF-SB(100μg·mL^(-1))+小干扰RNA对照(si-con,200nmol·L^(-1))组、TF-SB(100μg·mL^(-1))+MIIP小干扰RNA(si-MIIP,200nmol·L^(-1))组,转染质粒后TF-SB处理24h,CCK-8法检测细胞存活率;流式细胞术检测细胞凋亡;Western blotting法检测B细胞淋巴瘤-x1(Bcl-x1)、裂解的半胱氨酸天冬氨酸蛋白酶(cleavedCaspase-3)、MIIP表达;不同照射量X射线(0、2、4、6、8 Gy)照射后,克隆形成实验检测细胞存活率,并绘制单击多靶模型拟合曲线,Western blotting法检测γ-H2AX蛋白表达.结果与TF-SB 0μg·mL^(-1)组比较,TF-SB25、50、100、200μg·mL^(-1)组AGS细胞存活率显著降低(P<0.05);与对照组及溶剂对照组比较,TF-SB 100μg·mL^(-1)组AGS细胞凋亡率显著升高(P<0.05),Bcl-x1蛋白表达显著降低(P<0.05),cleaved Caspase-3和MIIP蛋白表达显著升高(P<0.05);放疗后,TF-SB组细胞存活率显著降低(P<0.05),γ-H2AX蛋白表达显著升高(P<0.05),放疗敏感性增加.与pcDNA组比较,pcDNA-MIIP组AGS细胞凋亡率显著增加,细胞存活率显著降低,Bcl-x1蛋白表达显著降低,cleaved Caspase-3蛋白表达增加(P<0.05);放疗后,pcDNA-MIIP组细胞存活率显著降低(P<0.05),γ-H2AX蛋白表达显著升高(P<0.05),敏感性增加.与TF-SB+si-con组比较,TF-SB+si-MIIP组AGS细胞凋亡率显著降低,细胞存活率显著增加,Bcl-x1蛋白表达显著增加,cleaved Caspase-3蛋白表达显著降低(P<0.05);放疗后,TF-SB+si-MIIP组细胞存活率显著升高(P<0.05),γ-H2AX蛋白表达显著降低(P<0.05),敏感性降低.结论TF-SB通过上调MObjective To investigate the effect and molecular mechanism of total flavonoids of Scutellaria barbata(TF-SB) on the proliferation, apoptosis and radiosensitivity of gastric cancer cell AGS. Methods The cell counting kit(CCK-8) method was used to detect the effects of different concentrations(0, 25, 50, 100, 200 μg·mL^(-1)) of TF-SB on the survival of gastric cancer cells to determine the experimental concentration. AGS was divided into control group, solvent control group(methanol), TF-SB(100 μg·mL^(-1)),pcDNA(200 nmol·L^(-1)), pcDNA-MIIP(200 nmol·L^(-1)), TF-SB(100 μg·mL^(-1)) + si-con(200 nmol·L^(-1)), TF-SB(100 μg·mL^(-1)) + siMIIP(200 nmol·L^(-1)) group. Cell viability was detected by CCK-8 method. Flow cytometry was used to detect apoptosis. Clone formation test was used to detect the cell survival fraction after irradiation with different rays, and a one-click multi-target model was fitted to fit the curve. Western blotting was used to detect B cell lymphoma-xl(Bcl-xl), cleaved Caspase-3, migration invasion inhibitor protein(MIIP), γ-H2AX protein expression. Results Compared with TF-SB 0 μg·mL^(-1)group, the survival rate of AGS cells in TF-SB 25, 50, 100 and 200 μg·mL^(-1)groups was significantly decreased(P < 0.05). Compared with control group and solvent control group, the apoptosis rate of AGS cells in TF-SB 100 μg·mL^(-1)group was significantly increased(P < 0.05), the expression of Bcl-XL protein was significantly decreased(P < 0.05), and the expression of Cleaved Caspase-3 and MIIP protein was significantly increased(P < 0.05). After radiotherapy, cell survival rate of TF-SB group was significantly decreased(P < 0.05), γ-H2AX protein expression was significantly increased(P < 0.05), and radiotherapy sensitivity was increased. Compared with the pcDNA group, the apoptosis rate of AGS cells in the pcDNA-MIIP group was significantly increased, the cell survival rate was significantly decreased,the expression of Bcl-XL protein was significantly decreased, and the expression of Cleaved

关 键 词：半枝莲总黄酮 人胃腺癌细胞AGS 迁移侵袭抑制蛋白(MIIP) 细胞增殖 凋亡 放疗敏感性 

分 类 号：R285.5[医药卫生—中药学]