Hong Gyeong Eun, Lee Ho Jeong, Kim Jin A, Yumnam Silvia, Raha Suchismita, Venkatarame Gowda Saralamma Venu, Heo Jeong Doo, Lee Sang Joon, Kim Eun Hee, Won Chun Kil, Kim Gon Sup
Research Institute of Life Science and College of Veterinary Medicine, Gyeongsang National University, Gazwa, Jinju 660-701, Republic of Korea.
Department of Physical Therapy, International University of Korea, Jinju 660-759, Republic of Korea.
Int J Oncol. 2017 Feb;50(2):575-586. doi: 10.3892/ijo.2016.3816. Epub 2016 Dec 22.
Citrus platymamma Hort.et Tanaka is an indigenous fruit of Jeju island in Korea. In this study the bioactivity of C. platymamma flavonoids were evaluated on human hepatoma Hep3B cell lines. Eleven flavonoids were identified from the peels of C. platymamma Hort.et Tanaka through high-performance liquid chromatography-Tandem mass spectrometry and the anticancer effect of these C. platymamma flavonoids on human hepatoma Hep3B were studied. Chromatin condensation was observed in Hep3B cells treated with C. platymamma flavonoids. DNA fragmentation was confirmed through agarose gel electrophoresis and TUNEL assay. An increase in the total apoptotic cells and G2/M cell cycle arrest with decreased protein expression of CDC25C, CDK1, cyclin B1 and p21 were observed in Hep3B cells treated with flavonoids of C. platymamma. Further, protein expression of Bcl-XL, Bax, caspase-3 and -9 were also modulated by C. platymamma flavonoids treatment indicating that cell death is through intrinsic apoptotic pathway. Moreover, C. platymamma flavonoids also regulated the phosphorylation of MAPKs, PI3K, and Akt in Hep3B cells. Relevant to inhibiting metastasis, C. platymamma treatment reduced wound closure of Hep3B cells and the protein expression of matrix metalloproteinase-2 and -9 were reduced in C. platymamma treated cells. The results show that C. platymamma flavonoids induce cell cycle arrest and apoptosis following activation of MAPKs and suppression of PI3K/Akt pathway which eventually inhibits cell migration in Hep3B cells. The finding provides evidence on biochemical activities of C. platymamma Hort.et Tanaka, which would be an essential agent for hepatocellular carcinoma (HCC) treatment.
宽皮柑橘是韩国济州岛的本土水果。在本研究中，对宽皮柑橘类黄酮对人肝癌Hep3B细胞系的生物活性进行了评估。通过高效液相色谱-串联质谱法从宽皮柑橘的果皮中鉴定出11种类黄酮，并研究了这些宽皮柑橘类黄酮对人肝癌Hep3B的抗癌作用。在用宽皮柑橘类黄酮处理的Hep3B细胞中观察到染色质浓缩。通过琼脂糖凝胶电泳和TUNEL测定法确认了DNA片段化。在用宽皮柑橘类黄酮处理的Hep3B细胞中，观察到总凋亡细胞增加，G2/M细胞周期停滞，同时CDC25C、CDK1、细胞周期蛋白B1和p21的蛋白表达降低。此外，Bcl-XL、Bax、caspase-3和-9的蛋白表达也受到宽皮柑橘类黄酮处理的调节，表明细胞死亡是通过内源性凋亡途径发生的。此外，宽皮柑橘类黄酮还调节了Hep3B细胞中MAPKs、PI3K和Akt的磷酸化。与抑制转移相关，宽皮柑橘处理减少了Hep3B细胞的伤口闭合，并且在宽皮柑橘处理的细胞中基质金属蛋白酶-2和-9的蛋白表达降低。结果表明，宽皮柑橘类黄酮在激活MAPKs和抑制PI3K/Akt途径后诱导细胞周期停滞和凋亡，最终抑制Hep3B细胞的迁移。这一发现为宽皮柑橘的生化活性提供了证据，宽皮柑橘可能是治疗肝细胞癌（HCC）的重要药物。
