Abstract：
BACKGROUND: Porcupine quills, a by-product of porcupine pork, are rich in keratin, which is an excellent source of bioactive peptides. The objective of this study was to investigate the underlying mechanism of anti-proliferation effect of porcupine quills keratin peptides (PQKPs) on MCF-7 cells.
RESULTS: Results showed that PQKPs induced MCF-7 cells apoptosis by significantly decreasing the secretion level of anti-apoptosis protein Bcl-2 and increasing the secretion levels of pro-apoptosis proteins Bax, cytochrome c, caspase 9, caspase 3 and PARP. PQKPs also arrested the cell cycle at G0/G1 phase via remarkably reducing the protein levels of CDK4 and enhancing the protein levels of p53 and p21. High-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) analysis identified nine peptides with molecular weights less than 1000 Da in PQKPs. Molecular docking results showed that TPGPPT and KGPAC identified from PQKPs could bind with p53 mutant and Bcl-2 protein by conventional hydrogen bonds, carbon hydrogen bonds and van der Waals force. Furthermore, the anti-proliferation impact of synthesized peptides (TPGPPT and KGPAC) was shown in MCF-7 cells.
CONCLUSIONS: These findings indicated that PQKPs suppressed the proliferation of MCF-7 breast cancer cells by triggering apoptosis and G0/G1 cell cycle arrest. Moreover, the outcome of this study will bring fresh insights into the production and application of animal byproducts. © 2023 Society of Chemical Industry.
RESULTS: Results showed that PQKPs induced MCF-7 cells apoptosis by significantly decreasing the secretion level of anti-apoptosis protein Bcl-2 and increasing the secretion levels of pro-apoptosis proteins Bax, cytochrome c, caspase 9, caspase 3 and PARP. PQKPs also arrested the cell cycle at G0/G1 phase via remarkably reducing the protein levels of CDK4 and enhancing the protein levels of p53 and p21. High-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) analysis identified nine peptides with molecular weights less than 1000 Da in PQKPs. Molecular docking results showed that TPGPPT and KGPAC identified from PQKPs could bind with p53 mutant and Bcl-2 protein by conventional hydrogen bonds, carbon hydrogen bonds and van der Waals force. Furthermore, the anti-proliferation impact of synthesized peptides (TPGPPT and KGPAC) was shown in MCF-7 cells.
CONCLUSIONS: These findings indicated that PQKPs suppressed the proliferation of MCF-7 breast cancer cells by triggering apoptosis and G0/G1 cell cycle arrest. Moreover, the outcome of this study will bring fresh insights into the production and application of animal byproducts. © 2023 Society of Chemical Industry.
摘要：
背景：豪猪羽毛，豪猪猪肉的副产品，富含角蛋白，这是生物活性肽的极好来源。本研究的目的是探讨豪猪毛笔角蛋白肽（PQKPs）对MCF-7细胞的抗增殖作用的潜在机制。
结果：结果表明，PQKPs通过显著降低抗凋亡蛋白Bcl-2的分泌水平和增加促凋亡蛋白Bax的分泌水平，诱导MCF-7细胞凋亡，细胞色素c,caspase9、caspase3和PARP。PQKP还通过显着降低CDK4的蛋白质水平并增强p53和p21的蛋白质水平将细胞周期阻滞在G0/G1期。高效液相色谱-串联质谱（HPLC-MS/MS）分析鉴定了PQKP中分子量小于1000Da的9种肽。分子对接结果表明，从PQKPs中鉴定出的TPGPPT和KGPAC可以通过常规氢键与p53突变体和Bcl-2蛋白结合，碳氢键和范德华力。此外，在MCF-7细胞中显示合成肽(TPGPPT和KGPAC)的抗增殖作用。
结论：这些发现表明，PQKPs通过触发凋亡和G0/G1细胞周期阻滞来抑制MCF-7乳腺癌细胞的增殖。此外，这项研究的结果将为动物副产品的生产和应用带来新的见解。©2023化学工业学会。
结果：结果表明，PQKPs通过显著降低抗凋亡蛋白Bcl-2的分泌水平和增加促凋亡蛋白Bax的分泌水平，诱导MCF-7细胞凋亡，细胞色素c,caspase9、caspase3和PARP。PQKP还通过显着降低CDK4的蛋白质水平并增强p53和p21的蛋白质水平将细胞周期阻滞在G0/G1期。高效液相色谱-串联质谱（HPLC-MS/MS）分析鉴定了PQKP中分子量小于1000Da的9种肽。分子对接结果表明，从PQKPs中鉴定出的TPGPPT和KGPAC可以通过常规氢键与p53突变体和Bcl-2蛋白结合，碳氢键和范德华力。此外，在MCF-7细胞中显示合成肽(TPGPPT和KGPAC)的抗增殖作用。
结论：这些发现表明，PQKPs通过触发凋亡和G0/G1细胞周期阻滞来抑制MCF-7乳腺癌细胞的增殖。此外，这项研究的结果将为动物副产品的生产和应用带来新的见解。©2023化学工业学会。
