DMDD对射频消融不完全肝癌细胞的抑制作用研究

唐立博; 范氏泰和; 何勇飞; 梁天仪; 莫书天; 韩箫; 农莹丹; 杨子叶; 卢春苗; 韩创业; 罗小玲

doi:10.16190/j.cnki.45-1211/r.2023.05.003

DMDD对射频消融不完全肝癌细胞的抑制作用研究

Inhibitory effect of DMDD on hepatocellular carcinoma cells with insufficient radiofrequency ablation

摘要

摘要: 目的：研究杨桃根提取物2-十二烷基-6-甲氧基-2, 5-二烯-1, 4-环己二酮（DMDD）对射频消融不完全人肝癌细胞HuH-7、Hep3B 模型的抑制作用。方法：经47 ℃水浴处理人肝癌细胞HuH-7、Hep3B 构建射频消融不完全细胞模型，采用细胞增殖实验（CCK-8）检测野生型HuH-7，Hep3B 细胞（对照组）和射频消融不完全模型细胞（模型组）的增殖能力，划痕实验检测细胞迁移能力，细胞侵袭实验（Transwell）检测细胞侵袭能力。CCK-8实验检测DMDD 对射频消融不完全肝癌细胞HuH-7，Hep3B的半数抑制浓度。选取3 种DMDD 浓度梯度，分别为DMDD 高浓度组（10 μmol/L，DMDD H 组）、DMDD 中浓度组（5 μmol/L，DMDD M 组）和DMDD 低浓度组（2.5 μmol/L，DMDD L 组），检测DMDD 干预后射频消融不完全肝癌细胞模型的增殖、迁移、侵袭能力；蛋白质印迹法（western blotting）检测DMDD 干预前后射频消融不完全肝癌细胞模型的METTL3 蛋白表达水平。结果：与对照组野生型HuH-7，Hep3B 细胞比较，射频消融不完全肝癌细胞HuH-7，Hep3B 的增殖、迁移、侵袭能力明显增强（P< 0.05），且METTL3蛋白表达水平上调（P< 0.05）；DMDD 干预后，DMDD H 组与未加药处理的对照组比较，在增殖、迁移、侵袭水平受到显著抑制，且METTL3 蛋白表达水平下调（P< 0.05）。结论：成功建立了射频消融不完全HuH-7，Hep3B 肝癌细胞模型，其具有较强的增殖、迁移、侵袭能力；DMDD 能够抑制射频消融不完全肝癌细胞的增殖、迁移、侵袭等恶性生物学特性，具有抗肿瘤作用。

Abstract: Objective:To study the inhibitory effect of 2-dodecyl-6-methoxy-2, 5-diene-1, 4-cyclohexanedione(DMDD), isolated from the root of Averrhoa Carambola L., on the HuH-7 and Hep3B model of human hepatocel-lular carcinoma cells with insufficient radiofrequency ablation.Methods:An insufficient radiofrequency ab-lation cell model was constructed by treating human hepatocellular carcinoma cells HuH-7 and Hep3B with a 47℃water bath.Cell proliferation assay (CCK-8) was performed to detect the proliferation ability of wild-type HuH-7, Hep3B cells (control group) and insufficient ra-diofrequency ablation model cells(model group), the migration ability of cells was detected by scratch assay, and the invasion ability of cells was detected by cell invasion assay (Transwell).CCK-8 assay was used to detect the half-inhibitory concentration of DMDD on hepatocellular carcinoma cells HuH-7 and Hep3B with insufficient ra-diofrequency ablation.Three DMDD concentration gradients were selected: high-concentration DMDD group(10 μmol/L, DMDD H group), medium-concentration DMDD group (5 μmol/L, DMDD M group) and low-con-centration DMDD group (2.5 μmol/L DMDD L group).The proliferation, migration and invasion abilities of in-sufficient radiofrequency ablation cell model were detected after DMDD intervention.Western blotting assay was used to measure METTL3 protein expression levels in the insufficient radiofrequency ablation cell model before and after DMDD intervention.Results:Compared with the wild-type HuH-7 and Hep3B cells in the control group, the proliferation, migration and invasion abilities of hepatocellular carcinoma cells HuH-7, Hep3B with in-sufficient radiofrequency ablation were significantly enhanced (P< 0.05), and the expression level of METTL3 protein was up-regulated (P< 0.05).After DMDD intervention, the proliferation, migration, and invasion levels were significantly inhibited and METTL3 protein expression level was down-regulated in the DMDD H group compared with the control group (P< 0.05).Conclusion:A HuH-7 and Hep3B hepatocellular carcinoma cell model with insufficient radiofrequency ablation is successfully established, with strong proliferation, migration and invasion abilities.DMDD can inhibit the proliferation, migration, invasion and other malignant biological properties of hepatocellular carcinoma cells with insufficient radiofrequency ablation, and has anti-tumor effect.

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