Luo Yang-Xin, Peng Bao-Ying, Chen Zheng-Hao, Xiong Xi-Kun, Huang Jun-Ming, Chen Mei-Fen, Wang Feng-Yan, Li Xin, Wang Jian-Ning
Department of Oral and Maxillofacial Surgery, Hospital of Stomatology, Guanghua School of Stomatology, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Stomatology, 56, Ling Yuan Xi Road, Guangzhou, Guangdong Province, P.R. China, 510055.
Guangdong Provincial Center for Disease Control and Prevention, 160 Qunxian Road, Dashi, Panyu District, Guangzhou, Guangdong Province, P.R. China, 511430.
Anticancer Agents Med Chem. 2023;23(4):432-439. doi: 10.2174/1871520622666220615121525.
The study aims to investigate the combined effects of chrysin and cisplatin on hepatoma(HepG2) cell lines in vivo and in vitro.
Studies have suggested that chrysin can enhance the sensitivity of tumor cells to apoptosis. Drug resistance in tumor cells reduced the effectiveness of chemotherapy drugs such as cisplatin. We investigated whether the combination of chrysin and cisplatin can induce more apoptosis than chrysin alone and cisplatin alone.
HepG2 cells were pretreated with chrysin for 2 h, followed by the addition of cisplatin for another 24 h. The morphologic changes were observed under inverted microscope and the cell viability was measured using the MTT test. The protein and cleavage of caspase-3,8,9, PARP, and cFLIP were determined by Western blotting.
The cell viability of the HepG2 cell can be reduced by the combination of chrysin pretreatment for 2 h and cisplatin addition for 24 h; Caspase-3,8,9 and PARP were cleaved after 12 h treatment with chrysin and cisplatin; Pancaspase inhibitor, Z-VAD-fmk, could reverse the apoptosis induced by chrysin and cisplatin in HepG2 cells; cFLIP was down-regulated by the combination of chrysin and cisplatin, and could be reversed by Z-VAD-fmk; the xenografted HepG2 cells formed a tumor in one week; At the end of the experiment, there were significant differences in relative tumor volume (RTV) and relative tumor proliferation rate between the combined group and the control group, the chrysin group and the cisplatin group; Western blotting showed that the levels of PARP, cFLIP, and caspase-3 proteins in isolated tumor tissues also decreased under the combined action of chrysin and cisplatin.
The combination of chrysin and cisplatin induces apoptosis of hepatic tumor in vivo and in vitro. It downregulates cFLIP and then activates caspase-8, which triggers caspase-mediated apoptosis of HepG2 cell.
本研究旨在探讨白杨素与顺铂联合应用对肝癌（HepG2）细胞系的体内外联合作用。
研究表明，白杨素可增强肿瘤细胞对凋亡的敏感性。肿瘤细胞中的耐药性降低了顺铂等化疗药物的疗效。我们研究了白杨素与顺铂联合应用是否比单独使用白杨素和顺铂诱导更多的细胞凋亡。
用白杨素预处理HepG2细胞2小时，然后再加入顺铂处理24小时。在倒置显微镜下观察形态学变化，并用MTT试验测定细胞活力。通过蛋白质印迹法测定caspase-3、8、9、PARP和cFLIP的蛋白及裂解情况。
白杨素预处理2小时后再加入顺铂处理24小时可降低HepG2细胞的活力；用白杨素和顺铂处理12小时后，Caspase-3、8、9和PARP被裂解；泛半胱天冬酶抑制剂Z-VAD-fmk可逆转白杨素和顺铂诱导的HepG2细胞凋亡；白杨素与顺铂联合应用可下调cFLIP，且可被Z-VAD-fmk逆转；接种的HepG2细胞在一周内形成肿瘤；实验结束时，联合组与对照组、白杨素组和顺铂组之间的相对肿瘤体积（RTV）和相对肿瘤增殖率存在显著差异；蛋白质印迹法显示，在白杨素和顺铂的联合作用下，分离的肿瘤组织中PARP、cFLIP和caspase-3蛋白水平也降低。
白杨素与顺铂联合应用可在体内外诱导肝肿瘤细胞凋亡。它下调cFLIP，然后激活caspase-8，从而触发caspase介导的HepG2细胞凋亡。
