In vitro Study on the Periplaneta americana Polypepticde PAE2 Reversing Multi-Drug Resistance of BEL-7402/5-FU Based on AMPK-Regulated Autophagy

Abstract

Abstract:

〔Abstract〕 Objective： To investigate the effects of the Periplaneta americana polypeptide PAE2 on the multi-drug resistance of BEL-7402/5-FU cells based on adenosine monophosphate-activited protein kinase（AMPK） regulation of autophagy. Methods： Human hepatocellular carcinoma sensitive cell line BEL-7402 and human hepatocellular carcinoma drug-resistant cell line BEL-7402/5-FU were used as the research objects. The action time and concentration of AICAR were determined by methyl thiazolyl tetrazolium（MTT） colorimetry assay. AMPK content in cells was determined by enzyme linked immunosorbent assay （ELISA）. Real-time polymerase chain reaction （real-time PCR） was used to detect AMPK mRNA expression level in cells. Real-time PCR was used to detect the mRNA expression levels of autophagy related protein 5（ATG5）， autophagy related protein 7（ATG7）， phosphoinositide-3-kinase class 3（PIK3C3）, Beclin 1，microtubuler-associated protein 1 light chain 3-Ⅱ（LC3-Ⅱ）， P62 and enhancer of Zeste homolog 2（EZH2）. The expression of LC3-Ⅱ was observed by laser scanning confocal microscope. The expression of P62 protein was detected by immunocytochemistry（ICC）. Results： Periplaneta americana polypeptide PAE2 can reduce the expression of AMPK and its mRNA， greatly reduce the expression of related autophagy proteins ATG5 mRNA， ATG7 mRNA， PIK3C3 mRNA， Beclin 1 mRNA， and LC3-Ⅱ mRNA（P<0.05）， and significantly increase the expression of P62 mRNA和EZH2 mRNA（P<0.05）， increase the expression of P62 protein（P<0.05）. Conclusion： Periplaneta americana polypeptide PAE2 can reverse the multi-drug resistance of BEL-7402/5-FU cells by inhibiting AMPK so as to inhibit autophagy.

Key words:

"> ">〔Key words">〕 Periplaneta americana polypeptide">； adenosine monophosphate-activited protein kinase">；autophagy">；multi-drug resistance

CLC Number:

Ou Ling, Li Cailin, Li Yinrui, Zhou Jie, Shan Wanxin, Wu Dingyu, Peng Fang. In vitro Study on the Periplaneta americana Polypepticde PAE2 Reversing Multi-Drug Resistance of BEL-7402/5-FU Based on AMPK-Regulated Autophagy[J]. Journal of Dali University, 2022, 7(4): 1-7.

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