美洲大蠊多肽PAE2逆转BEL-7402/5-FU细胞多药耐药性的作用及机制研究

Effect and Mechanism of the Periplaneta American Polypeptide PAE2 on Reversing Multi-Drug Resistance of BEL-7402/5-FU Cells

本研究探讨了美洲大蠊多肽PAE2逆转人肝细胞肿瘤耐药细胞株BEL-7402/5-FU的多药耐药性作用及机制。以人肝细胞肿瘤敏感细胞株BEL-7402、人肝细胞肿瘤耐5-氟尿嘧啶(5-FU)细胞株BEL-7402/5-FU和Balb/c-nude小鼠为研究对象,采用噻唑蓝比色分析法检测BEL-7402/5-FU细胞的多药耐药性和交叉耐药性,通过激光共聚焦显微镜观察阿霉素在BEL-7402/5-FU细胞中的累积。使用实时荧光定量聚合酶链式反应和免疫细胞化学染色法检测BEL-7402/5-FU细胞中P-糖蛋白1(P-glycoprotein 1,P-gp1)、多药耐药相关蛋白1(multi-drug resistance protein 1,MRP1)、肺耐药相关蛋白1(lung resistance protein 1,LRP1)以及乳腺癌耐药蛋白1(breast cancer resistance protein 1,BCRP1)的mRNA和蛋白的相对表达量,确定美洲大蠊多肽PAE2逆转多药耐药性的机制。此外,在Balb/c-nude小鼠中,建立BEL-7402/5-FU细胞的腋下移植瘤模型,观察美洲大蠊多肽PAE2对腋下移植瘤小鼠的免疫器官、肝肾功能、肿瘤抑制与诱导肿瘤细胞调亡、调节肿瘤组织多药耐药相关蛋白等环节的影响。结果显示,美洲大蠊多肽PAE2能有效逆转BEL-7402/5-FU细胞的多药耐药性和交叉耐药性,作用机制可能与下调多药耐药相关蛋白的表达水平有关,降低了多药耐药细胞对抗肿瘤药物的外排能力、影响抗肿瘤药物分子代谢,从而达到逆转人肝细胞癌多药耐药的作用。

The effect and mechanism of periplaneta American polypeptide PAE2 in reversing the multi-drug resistance of hepatocellualar carcinoma resistant BEL-7402/5-FU cells were studied in this paper.Hepatocellualar carcinoma sensitive cells of BEL-7402,hepatocellualar carcinoma resistant 5-fluorouracil(5-FU)cells of BEL-7402/5-FU and Balb/c-nude mice were used as the research objects.MTT method was used to detect the multi-drug and crossdrug resistance of BEL-7402/5-FU cells,and the accumulation of doxorubicin in BEL-7402/5-FU cells was observed by laser confocal microscope.In order to determine the mechanism of the periplaneta American polypeptide PAE2 in reversing multi-drug resistance,real-time fluorescent quantitative polymerase chain reaction and immunocytochemical staining method were used to detect the mRNA and protein relative expression of P-glycoprotein 1(P-gp1),multi-drug resistance protein 1(MRP1),lung resistance protein 1(LRP1)and breast cancer resistance protein 1(BCRP1)in BEL-7402/5-FU cells.In addition,an axillary tumor model of BEL-7402/5-FU cells was established in Balb/c-nude mice to observe the effects of periplaneta American polypeptide PAE2 on immune organs,liver and kidney function,tumor inhibition and induction of tumor cell apoptosis,as well as regulation of multi-drug resistance related proteins in tumor tissues.The results showed that the periplaneta American polypeptide PAE2 could effectively reverse the multi-drug and cross-drug resistance of BEL-7402/5-FU cells.The mechanism might be related to the down-regulation of the expression of multi-drug resistance related proteins,which reduced the efflux ability of multi-drug resistant cells on anti-tumor drugs and intervene molecular metabolism of anti-tumor drugs,reversing the multi-drug resistance of hepatocellualar carcinoma finally.