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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 observe the effect of Shenqi Yiliu Fang (Yiliufang) serum on MKN-45 proliferation and invasion and metastasis, and to explore its mechanism. Methods: MTT assay was used to detect cell viability. Trans-well and scratch assays were performed to evaluate cell invasion and migration. Real-time PCR was used to detect COX- 2), and human phosphatase (PTEN) mRNA on human chromosome 10. The expressions of COX-2 and PTEN protein were detected by immunohistochemistry. Results: The inhibitory rates of Shenqi Yiliu Decoction-containing serum (2.8 g · m L-1) were 31.25%, 33.09% and 34.50% after 24, 48 and 72 h intervention respectively. The invasion, migration and migration of cells decreased by 36.18%, 49.46% and 36.46%, respectively. The expression of COX-2 mRNA decreased by 90% and PTEN increased by 89.72% Compared with the blank serum group, the expression of COX-2 protein in Shenqi Yiliu prescription group was weakened; the expression of PTEN protein was enhanced (P <0.05). Conclusion: Shenqi Yiliufang serum can inhibit the proliferation of MKN-45 cells by decreasing the expression of COX-2, PTEN mRNA and protein, and weakening the invasion and metastasis of MKN-45 cells.