黄芩素对人胃癌SGC7901细胞迁移及侵袭的影响及其作用机制
Effects of baicalein on the migration and invasion of human gastric carcinoma SGC7901 cells and its mechanism

摘要：目的 观察黄芩素对人胃癌SGC7901细胞迁移及侵袭的影响及其作用机制。方法 以人胃癌SGC7901细胞为研究对象，分别加入20mL的培养液(对照组)或20mL(50、100、150mol/L)黄芩素溶液处理细胞48h，细胞划痕试验检测不同剂量黄芩素对人胃癌SGC7901细胞迁移的影响，Transwell侵袭实验检测黄芩素对细胞侵袭能力的改变；比色法检测细胞胰蛋白酶活性，用Western blot方法检测活化的蛋白酶激活受体-2(protease-activated receptor-2, PAR-2)蛋白表达水平，RT-PCR法检测基质金属蛋白酶-2(matrix metalloproteinases-2, MMP-2)mRNA及基质金属蛋白酶-9(matrix metalloproteinases-9, MMP-9)mRNA水平。结果 与对照组比较，各治疗组人胃癌SGC7901细胞迁移及侵袭的能力、胰蛋白酶活性、活化的PAR-2、MMP-2 mRNA及MMP-9 mRNA表达水平均显著下降(P<0.05)，呈剂量依赖性。活化的PAR-2与MMP-2 mRNA及MMP-9 mRNA表达水平呈正相关关系(r=0.564，P<0.05；r=0.581，P<0.05)。结论 PAR-2活性、MMP-2及MMP-9 mRNA表达与胃癌的发生、发展密切相关，活化的PAR-2可通过上调MMP-2及MMP-9 mRNA的表达而参与癌细胞迁移及侵袭，黄芩素能显著抑制人胃癌SGC7901细胞的PAR-2活性和MMP-2及MMP-9 mRNA表达，从而降低癌细胞迁移及侵袭能力，这与其抑制胰蛋白酶活性有密切关系。
ABSTRACT: Objective To observe the effects of baicalein on the migration and invasion of human gastric carcinoma SGC7901 cells and its mechanism. Methods Human gastric carcinoma SGC7901 cells cultured in vitro were respectively treated with 20mL culture medium (control group), 50mol/L baicalein, 100mol/L baicalein, and 150mol/L baicalein for 48 hours. Wound healing and Transwell assay were used to determine the inhibitory effects of baicalein on cell migration and invasion in human gastric carcinoma SGC7901 cells respectively. The typsin activity of cells was detected by photocolorimetric method, the protein expression of active protease-activated receptor-2 (PAR-2) was detemined by Western blotting; the matrix metalloproteinases-2 (MMP-2) mRNA, matrix metalloproteinases-9 (MMP-9) mRNA expressions were measured with RT-PCR. Results Compared with those in control group, the abilities of cells migration and invasion and the typsin activity, active PAR-2, MMP-2 mRNA and MMP-9 mRNA expressions were dose-dependently decreased in three baicalein groups (P<0.01). There were obvious positive correlations among active PAR-2, MMP-2 mRNA and MMP-9 mRNA expressions (r=0.564, P<0.05; r=0.681, P<0.05). Conclusion PAR-2 activity, MMP-2 mRNA and MMP-9 mRNA expressions are closely correlated with the pathogenesis of gastric carcinoma. Active PAR-2 may play an important role in carcinoma cells migration and invasion via upregulating MMP-2 mRNA and MMP-9 mRNA expressions. Baicalein can inhibit PAR-2 activity, MMP-2 mRNA and MMP-9 mRNA expressions obviously in human gastric carcinoma SGC7901 cells, thus decreasing the carcinoma cells migration and invasion, which is closely correlated to its inhibiting effects on typsin activity