黄芩素抑制胃癌HGC-27细胞上皮间充质转化进程研究

Baicalein inhibited the epithelial-to-mesenchymal transition process of gastric cancer HGC-27 cells

目的探讨黄芩素(baicalein,BAI)对胃癌HGC-27细胞增殖、周期和上皮间质转化(EMT)的影响及机制。方法将HGC-27细胞分成对照组和低、中、高剂量的黄芩素处理组,分别使用终浓度为0、5、15和25μmol·L^(-1)黄芩素处理48 h。噻唑蓝(MTT)法和平板克隆形成实验检测细胞增殖能力;划痕愈合实验和Transwell小室迁移实验检测细胞迁移能力;流式细胞术检测细胞周期;蛋白质印迹法检测周期、EMT相关蛋白的表达。结果对照组和低、中、高剂量实验组细胞抑制率分别为(0.97±9.62)%、(19.11±4.24)%、(30.44±8.50)%和(38.62±5.50)%;这4组48 h的横向迁移率分别为(81.44±0.31)%、(48.97±2.46)%、(21.45±2.47)%和(14.69±3.15)%;这4组纵向迁移率分别为(97.67±2.52)%、(84.33±4.04)%、(44.33±3.79)%和(13.33±3.06)%;这4组G0/G1期阻滞率分别为(23.84±2.95)%、(25.96±1.70)%、(29.11±3.32)%和(31.65±2.41)%;这4组上皮标志蛋白E-cadherin蛋白表达水平分别为0.97±0.14、0.17±0.22、1.36±0.32和1.99±0.23;Vimentin蛋白表达水平分别为1.00±0.23、0.82±0.13、0.50±0.18和0.25±0.12;这4组周期相关蛋白细胞周期蛋白依赖性激酶4(CDK4)蛋白表达水平分别为1.05±0.19、0.88±0.19、0.65±0.07和0.51±0.12;CDK6蛋白表达水平分别为0.90±0.18、0.63±0.18、0.57±0.07和0.37±0.03;CyclinB1蛋白相关表达水平分别为1.20±0.05、0.99±0.04、0.71±0.17和0.59±0.17;CyclinD1蛋白相关表达水平分别为1.47±0.24、1.31±0.36、0.82±0.37和0.37±0.07。低、中、高剂量实验组与对照组相比,差异均有统计意义(均P<0.05)。结论BAI可有效抑制胃癌HGC-27细胞的增殖和EMT进程,其机制与BAI调控周期及EMT相关蛋白表达水平相关。

Objective To investigate the effects of baicalein(BAI)on proliferation,cycle and epithelial-mesenchymal transformation(EMT)of gastric cancer HGC-27 cells and its mechanism.Methods HGC-27 cells were divided into control group,and experimental-L,-M,-H groups,treated with baicalein at final concentrations of 0,5,15 and 25μmol·L^(-1)for 48 h.The cell proliferation was detected by methy thiazolyl tetrazolium(MTT)assay and colony formation assay.The cel migration ability was detected by scratch healing assay and Transwel chamber migration assay.Cell cycle was detected by flow cytometry.The expressions of cycle and EMT related proteins were detected by Western blot.Results The cell inhibition rates were(0.97±9.62)%,(19.11±4.24)%,(30.44±8.50)%and(38.62±5.50)%in control group,experimental-L,experimental-M and experimental-H group,respectively.The transverse mobility at 48 h were(81.44±0.31)%,(48.97±2.46)%,(21.45±2.47)%and(14.69±3.15)%,respectively.The longitudinal mobility of the four groups were(97.67±2.52)%,(84.33±4.04)%,(44.33±3.79)%and(13.33±3.06)%,respectively.The G0/G1 block rates of the 4 groups were(23.84±2.95)%,(25.96±1.70)%,(29.11±3.32)%and(31.65±2.41)%,respectively.The expression levels of E-cadherin were 0.97±0.14,0.17±0.22,1.36±0.32 and 1.99±0.23,respectively.Vimentin protein expression level were 1.00±0.23,0.82±0.13,0.50±0.18 and 0.25±0.12,respectively.The expression levels of cyclin-dependent kinase 4(CDK4)were 1.05±0.19,0.88±0.19,0.65±0.07 and 0.51±0.12,respectively.CDK6 protein expression levels were 0.90±0.18,0.63±0.18,0.57±0.07 and 0.37±0.03,respectively.CyclinB1 protein related expression levels were 1.20±0.05,0.99±0.04,0.71±0.17 and 0.59±0.17,respectively.CyclinD1 protein related expression levels were 1.47±0.24,1.31±0.36,0.82±0.37 and0.37±0.07,respectively.Compared with the control group,there were statistically significant differences in experimental-L,-M,-H groups(all P<0.05).Conclusion BAI can effectively inhibit the proliferation and EMT process of gastric cancer HGC-27 cells,and the mechanism is related to the regulatory cycle of BAI and the expression level of EMT-related proteins.