美洲大蠊多肽PAE_(2)对人肝癌BEL-7402和BEL-7402/5-FU裸鼠皮下移植瘤侵袭转移的影响

Effect of Periplaneta americana polypeptide PAE_(2) on invasion and metastasis of human liver cancer BEL-7402 and BEL-7402/5-FU subcutaneously transplanted tumor in nude mice

目的 基于腺苷酸活化蛋白激酶靶点在动物水平上探讨美洲大蠊多肽PAE;逆转肝癌多药耐药细胞BEL-7402/5-FU的机制研究。方法 以Balb/c-nude裸鼠为实验对象,建立裸鼠皮下人肝癌BEL-7402和BEL-7402/5-FU细胞瘤模型(敏感肝癌细胞模型和耐药肝癌细胞模型)。将裸鼠分为正常组、敏感组(敏感肝癌细胞模型)、耐药组(耐药肝癌细胞模型),索拉菲尼组(耐药肝癌细胞模型,给予30 mg·kg^(-1)阳性药索拉菲尼)和高、中、低剂量实验组(耐药肝癌细胞模型,分别给予200,100,50 mg·kg^(-1)多肽PAE_(2))。计算裸鼠抑瘤率及脏器指数;以生化分析仪检测裸鼠生化指标;以苏木精-伊红(HE)染色法观察肿瘤组织及肝组织的病理学变化;以实时定量反转录聚合酶链反应(qRT-PCR)法检测肿瘤组织中侵袭转移相关因子mRNA表达水平。结果 敏感组、耐药组、索拉菲尼组和高、中、低3个剂量实验组MMP2表达水平分别为0.01±0.00,1.00±0.00,0.02±0.01,0.01±0.01,0.07±0.02和0.00±0.00;这6组的MMP9表达水平分别为0.01±0.02,1.00±0.00,0.18±0.08,0.01±0.00,0.04±0.01和0.00±0.00;这6组的4EBP1表达水平分别为0.23±0.08,1.00±0.00,1.80±0.30,0.20±0.06,0.66±0.16和0.12±0.06。PAE;组与耐药组相比,各项mRNA表达水平均明显下降,差异均有统计学意义(均P<0.01)。结论 美洲大蠊多肽PAE_(2)可通过腺苷酸活化蛋白激酶靶点影响裸鼠肿瘤细胞的侵袭转移因子,降低肿瘤组织中MMP2等mRNA的侵袭,从而抑制裸鼠肿瘤的生长。

Objective To investigate the mechanism of Periplaneta americana polypeptide PAE_(2)in reversing the multidrug resistance cell BEL-7402/5-FU of liver cancer based on the Adenosine 5′-monophosphate(AMP)-activated protein kinase target at the animal level. Methods Balb/c-nude nude mice were used as experimental subjects to establish subcutaneous human hepatoma BEL-7402 and BEL-7402/5-FU cell tumor models in nude mice(sensitive liver cancer cell model and drug-resistant liver cancer cell model). Nude mice were divided into normal group, sensitive group(sensitive liver cancer cell model), drug-resistant group(resistant liver cancer cell model), and sorafenib group (resistant liver cancer cell model,given the positive drug sorafenib 30 mg·kg^(-1)) and experimental-H,-M,-L groups (drug-resistant hepatocellular carcinoma cell model,given polypeptide PAE2200,100,50mg·kg^(-1)). The tumor inhibition rate and organ index of nude mice were calculated. The biochemical indexes of nude mice were detected by biochemical analyzer. The pathological changes of the tumor tissue and liver tissue were observed by hematoxylin-eosin (HE) staining. The mRNA expression levels of invasion and metastasis-related factors in tumor tissues were detected by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR).Results Periplaneta americana polypeptide PAE;on the subcutaneous xenograft tumor invasion and metastasis-related factors of nude mice in the sensitive group,drug-resistant group,sorafenib group and experimental-H,-M,-L groups,the MMP2 expression levels were 0. 01 ± 0. 01,1. 00 ± 0. 00,0. 02 ± 0. 01,0. 01 ± 0. 01,0. 07 ± 0. 02,0. 00 ± 0. 00;the MMP9 expression levels were 0. 01 ± 0. 02,1. 00 ± 0. 00,0. 18 ± 0. 08,0. 01 ± 0. 00,0. 04 ± 0. 01,0. 00 ± 0. 00;the 4EBP1 expression levels were 0. 23 ± 0. 08,1. 00 ± 0. 00,1. 80 ± 0. 30,0. 20 ± 0. 06,0. 66 ± 0. 16,0. 12 ± 0. 06. Compared between the drug-resistant group and experimental groups,the experimental group had a significant decrease in various mRNA expression factors, and the differences were statistically significant( all P < 0. 01). Conclusion Periplaneta americana polypeptide PAE;can affect the invasion and transfer factors of tumor cells in nude mice through Adenosine 5’-monophosphate( AMP)-activated protein kinase targets,reduce the invasion of MMP2 and other mRNA in tumor tissues,thereby inhibiting tumor growth in nude mice.