参芪抑瘤方抑制胃癌细胞MKN-45增殖和侵袭转移作用机制

Inhibitory Effect of Shenqi Yiliu Decoction on Proliferation and Metastasis of Gastric Cancer Cell Lines MKN-45

目的:观察参芪抑瘤方(抑瘤方)血清抑制MKN-45增殖和侵袭转移作用,并探讨其作用机制。方法:采用噻唑蓝(MTT)比色法检测细胞活力;Trans-well和划痕实验观察细胞侵袭与迁移能力;实时荧光定量PCR(Real-time PCR)技术检测环氧合酶-2(COX-2)和人第10号染色体缺失的磷酸酶(PTEN)mRNA表达;免疫组化检测细胞COX-2,PTEN蛋白表达。结果:参芪抑瘤方含药血清(2.8 g·m L-1)干预24,48,72 h后,抑制率为31.25%,33.09%,34.50%;参芪抑瘤方含药血清(2.8 g·m L-1)干预48 h后,细胞侵袭能力、转移能力和迁移能力分别下降了36.18%,49.46%,36.46%;COX-2 mRNA表达量下降90%,PTEN升高89.72%;与空白血清组比较,参芪抑瘤方组COX-2蛋白表达减弱;PTEN蛋白表达增强(P<0.05)。结论:参芪抑瘤方药物血清通过影响COX-2,PTEN mRNA和蛋白表达,抑制MKN-45细胞的增殖,减弱其侵袭转移能力。

Objective： To investigate the inhibitory effect and mechanisms of Shenqi Yiliu decoction（Yi Liu decoction） medicated serum on proliferation and metastasis of gastric cancer cell line MKN-45. Method：Methyl thiazolyl tetrazolium（MTT） was employed to test the viability of gastric cancer cell lines MKN-45. Transwell test and scratch test were used to detect the invasion and metastasis abilities. Real-time PCR technique was used to detect the expression of cyclooxygenase-2（COX-2） and phosphatase and tensin homolog deleted on chromosome ten（PTEN） mRNA. Immunocytochemical technique was used to test the protein expressions of COX-2and PTEN. Result： After MKN-45 cell were treated by high-dose Shenqi Yiliu decoction medicated serum（2. 8g·m L- 1） for 24 h,48 h and 72 h,the proliferation inhibition rates were 31. 25%,33. 09% and 34. 50%,respectively. After MKN-45 cell were treated by high-dose Shenqi Yiliu decoction medicated serum for 48 h,the abilities of invasion metastasis and migration decreased by 36. 18%,49. 46% and 36. 46%,respectively,the expressions of COX-2 mRNA decreased by 90%,and the expression of PTEN increased by 89. 72%. Compared with blank group,COX-2 protein expression was weakened in Shenqi Yiliu decoction medicated serum. PTEN protein expression was increased（P〈 0. 05）. Conclusion： Shenqi Yiliu decoction medicated serum inhibits the proliferation of MKN-45 cells and weakens the ability of invasion and metastasis by affecting the mRNA and protein expressions of COX-2 and PTEN.