JZUS - Journal of Zhejiang University SCIENCE

Journal of Zhejiang University SCIENCE B 2021 Vol.22 No.2 P.112-122

http://doi.org/10.1631/jzus.B2000446

Hydroxychavicol, a polyphenol from Piper betle leaf extract, induces cell cycle arrest and apoptosis in TP53-resistant HT-29 colon cancer cells

Author(s): Aiysvariyah RAJEDADRAM, Kar Yong PIN, Sui Kiong LING, See Wan YAN, Mee Lee LOOI
Affiliation(s): School of Biosciences, Taylor's University, Lakeside Campus, 47500 Subang Jaya, Malaysia; more
Corresponding email(s): meelee.looi@um.edu.my
Key Words: Piper betle, Hydroxychavicol (HC), Cell cycle, Apoptosis, c-Jun N-terminal kinase (JNK), P38 mitogen-activated protein kinase (MAPK)

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Abstract: This study aims to elucidate the antiproliferative mechanism of hydroxychavicol (HC). Its effects on cell cycle, apoptosis, and the expression of c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MAPK) in HT-29 colon cancer cells were investigated. HC was isolated from Piper betle leaf (PBL) and verified by high-performance liquid chromatography (HPLC), nuclear magnetic resonance (NMR), and gas chromatography-mass spectrometry (GC-MS). The cytotoxic effects of the standard drug 5-fluorouracil (5-FU), PBL water extract, and HC on HT-29 cells were measured after 24, 48, and 72 h of treatment. cell cycle and apoptosis modulation by 5-FU and HC treatments were investigated up to 30 h. Changes in phosphorylated JNK (pJNK) and P38 (pP38) MAPK expression were observed up to 18 h. The half maximal inhibitory concentration (IC₅₀) values of HC (30 μg/mL) and PBL water extract (380 μg/mL) were achieved at 24 h, whereas the IC₅₀ of 5-FU (50 μmol/L) was obtained at 72 h. cell cycle arrest at the G₀/G₁ phase in HC-treated cells was observed from 12 h onwards. Higher apoptotic cell death in HC-treated cells compared to 5-FU-treated cells (P<0.05) was observed. High expression of pJNK and pP38 MAPK was observed at 12 h in HC-treated cells, but not in 5-FU-treated HT-29 cells (P<0.05). It is concluded that HC induces cell cycle arrest and apoptosis of HT-29 cells, with these actions possibly mediated by JNK and P38 MAPK.

从花椒叶中提取的羟基胡椒酚可以诱导耐TP53的HT-29结肠癌细胞的细胞周期阻滞和凋亡

目的：阐明羟基胡椒酚（HC）的抗恶性细胞增生的机制。并研究HC对HT-29结肠癌细胞的细胞周期、凋亡及c-Jun氨基末端激酶（JNK）和P38丝裂原活化蛋白激酶（MAPK）表达的影响。
方法：从蒌叶（PBL）中分离出HC，经高效液相色谱（HPLC）、核磁共振（NMR）和气相色谱-质谱（GC-MS）进行检测。在处理24、48和72 h后，检测标准药物5-氟尿嘧啶（5-FU）、PBL水提物和HC对HT-29细胞的细胞毒性作用。检测30 h内5-FU和HC处理对细胞周期和凋亡的调控作用。同时检测18 h内磷酸化JNK（pJNK）和磷酸化P38（pP38）MAPK的表达变化。
结果：HC（30μg/ mL）和PBL水提物（380μg/ mL）的半数最大抑制浓度（IC₅₀）值在24 h时达到，而5-FU（50μmol/ L）的IC₅₀值在72 h时达到。从12h开始HC处理的细胞停滞在细胞周期的G₀/G₁期。与5-FU处理的细胞相比，HC处理的细胞凋亡率更高（P<0.05）。在HC处理的细胞中，pJNK和pP38 MAPK在12 h时出现高表达，而在5-FU处理的HT-29细胞中则没有（P<0.05）。
结论：由此可见，HC可诱导HT-29细胞的细胞周期阻滞和凋亡，这些作用可能由JNK和P38 MAPK介导。

关键词：蒌叶；羟基胡椒酚（HC）；细胞周期;细胞凋亡；c-Jun氨基末端激酶（JNK）；P38丝裂原活化蛋白激酶（MAPK）

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从花椒叶中提取的羟基胡椒酚可以诱导耐TP53的HT-29结肠癌细胞的细胞周期阻滞和凋亡

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