摘要
本研究为探究大黄酸对甲状腺乳头状癌细胞TPC-1增殖、侵袭及迁移的影响及其与Wnt/β-catenin信号通路的关系.将TPC-1细胞分为对照组、大黄酸低浓度组、大黄酸中浓度组、大黄酸高浓度组,另将TPC-1细胞分为对照组、大黄酸+SKL2001组(SKL2001为Wnt/β-catenin信号通路激活剂)、大黄酸+XAV939组(XAV939为Wnt/β-catenin信号通路抑制剂)、大黄酸组.采用CCK8法检测细胞增殖情况,采用Transwell试验检测细胞侵袭情况,采用划痕实验检测细胞迁移情况,采用Western blot法及RT-PCR法检测β-catenin和c-Myc蛋白及mRNA的表达情况.结果表明,大黄酸低浓度组、大黄酸中浓度组、大黄酸高浓度组均能有效抑制TPC-1细胞增殖,且大黄酸浓度越高、作用时间越长,TPC-1细胞抑制率越高(P＜0.05);大黄酸高浓度组细胞迁移数量、划痕愈合率低于大黄酸中浓度组、大黄酸低浓度组及对照组,差异显著(P＜0.05);大黄酸高浓度组β-catenin和c-Myc蛋白及mRNA表达量均低于对照组、大黄酸低浓度组及大黄酸中浓度组,差异显著(P＜0.05);大黄酸+SKL2001组TPC-1细胞抑制率低于大黄酸组,大黄酸+XAV939组TPC-1细胞抑制率高于大黄酸组,差异显著(P＜0.05);大黄酸+SKL2001组细胞迁移数量、划痕愈合率高于大黄酸组,大黄酸+XAV939组细胞迁移数量、划痕愈合率低于大黄酸组,差异显著(P＜0.05).本研究表明大黄酸可有效抑制甲状腺乳头状癌细胞增殖、侵袭及迁移,作用机制可能与其抑制Wnt/β-catenin信号通路有关.
Abstract
To investigate the effects of rhein on the proliferation,invasion and migration of thyroid papillary carcinoma cell line TPC-1 and its relationship with Wnt/β-catenin signaling pathway.TPC-1 cells were divided into control group,low-concentration rhein group,medium-concentration rhein group,and high-concentration rhein group.TPC-1 cells were also divided into control group,rhein+SKL2001 group(SKL2001 is an activator of Wnt/β-catenin signaling pathway),rhein+XAV939 group(XAV939 is an inhibitor of Wnt/β-catenin signaling pathway),and rhein group.Cell proliferation was detected by CCK8 method,cell invasion was determined by using the transwell assay,cell migration was detected by scratch test,and the expression of β-catenin and c-Myc protein and mRNA was detected by western blot and the RT-PCR method.The results showed that,the proliferation of TPC-1 cells in the low,medium,and high-concentration rhein groups was ef-fectively inhibited,and the higher the concentration and the longer the time,the higher the inhibitory rate of TPC-1 cells(P＜0.05).The number of cell migrations and the healing rate of scratches in the high-concentration rhein group was lower than that in the medium-concen-tration rhein group,low-concentration rhein group,and control group,with significant differences(P＜0.05).The expression levels of β-catenin and c-Myc protein and mRNA in the high-concentration rhein group were lower than those in the control group,low-concentration rhein group,and medium-concentration rhein group,with significant differences(P＜0.05).The inhibitory rate of TPC-1 cells in the rhein+SKL2001 group was lower than that in the rhein group,and the healing rate of scratches in the rhein+SKL2001 group was higher than that in the rhein group,with significant differences(P＜0.05).The inhibitory rate of TPC-1 cells in the rhein+XAV939 group was higher than that in the rhein group,and the number of cell migrations,the healing rate of scratches in the rhein+XAV939 group was lower than that in the rhein group,with significant differences(P＜0.05).This study showed that rhubarbic acid could effectively inhibit the proliferation,in-vasion and migration of papillary thyroid cancer cells,and the mechanism of action might be related to its inhibition of the Wnt/β-catenin signalling pathway.
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