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目的:探讨5-氟尿嘧啶(5-Fu)温热化疗对人低分化胃癌细胞(MKN45细胞)抑制的分子作用机制。方法:以MTT法测定MKN45细胞48h的IC50作为实验工作浓度,实验分对照组(C)、热疗组(H)、化疗组(D)、热化疗组(HD),检测细胞抑制率;应用逆转录-聚合酶链反应(RTPCR)观察表皮生长因子受体EGFR与原癌基因RAS mRNA的表达变化。结果:热、化疗均可抑制MKN45细胞系(P<0.05),热化疗抑制作用更明显(P<0.05);RT-PCR分析显示,热化疗组EGFRmRNA与RASmRNA表达下调最明显(P<0.05)。EGFR mRNA与RAS mRNA的表达呈明显的正相关性(r=0.870,P<0.05),细胞抑制率与EGFRmRNA表达水平间存在负相关关系(r=-0.956,P<0.05),细胞抑制率与RASmRNA表达水平间存在负相关关系(r=-0.923,P<0.05)。结论:化疗联合热疗能显著增加对MKN45细胞的体外抑制作用,其机制可能与下调EGFR与RAS的表达及二者相互作用有关。
Objective: To investigate the molecular mechanism of 5-fluorouracil (5-Fu) chemotherapy in inhibiting human poorly differentiated gastric cancer cells (MKN45). Methods: The IC50 of MKN45 cells for 48h was determined by MTT assay. The experimental groups were divided into experimental group (C), hyperthermia group (H), chemotherapy group (D) and thermochemotherapy group (HD) Expression of Epidermal Growth Factor Receptor EGFR and Proto - oncogene RAS mRNA in RTPCR Transcription - Transcription - Polymerase Chain Reaction. Results: Both heat and chemotherapy could inhibit the MKN45 cell line (P <0.05), and the inhibition of hyperthermia was more obvious (P <0.05). The RT-PCR analysis showed that the expression of EGFRmRNA and RASmRNA were significantly down-regulated . There was a significant positive correlation between the expression of EGFR mRNA and RAS mRNA (r = 0.870, P <0.05). There was a negative correlation between the expression of EGFR mRNA and EGFR mRNA (r = -0.956, P <0.05) There was a negative correlation between RASmRNA expression levels (r = -0.923, P <0.05). Conclusion: Chemotherapy combined with hyperthermia can significantly increase the inhibitory effect of MKN45 cells in vitro. The mechanism may be related to the down-regulation of EGFR and RAS expression and their interaction.