Quercetin上调Nm23-H1表达抑制肝细胞癌的侵袭转移

Quercetin suppresses the invasion and metastasis of hepatocellular carcinoma by upregulating Nm23-H1 expression

【摘要】目的研究quercetin和精胺调节Nm23-H1基因表达对肝癌侵袭转移能力的影响。方法RT—PCR和Western blot检测不同浓度和时间quereetin和精胺作用后SF7721细胞Nm23-H1mRNA和蛋白表达，Transwell和细胞黏附实验检测侵袭力和黏附力．并检测骨架蛋白α-tublin以及Factin的含量以及分布情况：裸鼠移植人肝癌模型腹腔注射相应药物，检测肝癌生长转移及Nm23-H1蛋白表达。结果Quercetin浓度及时间梯度处理细胞后Nm23-HImRNA（分别F=169．2，215．6，均P〈0．001）、蛋白表达（分别F=3530，2928，均P〈0．001，）以及黏附力（分别F=75．31，88．06，均P〈0．001）均高于对照组；侵袭力低于对照组（分别F=24．76，58．89，均P〈0．001）；骨架蛋白α-tublin及F—actin荧光强度均低于对照组，分布情况无明显变化；精胺浓度及时间梯度处理后各组细胞Nm23-HImRNA（分别F=283．1，623．5，均P〈0．001）和蛋白表达（分别F=4083，171，均P〈0．001）均低于对照组；侵袭力高于对照组（分别F=125．8，48．18，均P〈0．001）。Quercetin组、精胺组、对照组肝癌组织体积重量比为：2．07±0．90比6．84±2．15比7．02±2．30，三组肺转移率对比为0／8比4／8比5／8．quereetin组及精胺组Nm23-H1蛋白表达水平较对照组分别为上调及下调（P〈0．05）。结论Quereetin可上调肝癌细胞Nm23-H1基因表达，抑制肝癌侵袭转移能力。

Objective To investigate the effect of Nm23-H1 on the invasiveness and metastasis of hepatocellular carcinoma. Methods The expression of Nm23-H1 mRNA and protein in SF7721 cells were measured by RT-PCR or Western blot, respectively, cell invasiveness and cell adhesion were measured by transwell and cell adhesion assay, distribution and abundance of α-tublin and F-actin in cells were measured. Nude mouse model of human hepatoeellular carcinoma was intraperitoneally injected with specific drug, the growth, metastasis and expression of Nm23-H1 protein were measured. Results After the treatment by quercetin in concentration and time gradients, Nm23-H1 mRNA （respectively F = 169. 2, 215.6, all P 〈 0. 001 ） and protein expression level （ respectively F = 3 530, 2 928, all P 〈 0. 001 ）, adhesion ability （ respectively F = 75. 31, 88. 06,all P 〈0. 001 ） of cells were higher than those in control, the invasiveness （respectively F =24. 76, 58. 89, all P 〈 0. 001 ） of cells were weaker than those in control, the fluorescence intensity of F-actin and α-tublin were weaker than control. After the treatment by spennin, Nm23-H1 mRNA （respectively F =283. 1,623. 5,all P 〈 0. 001 ） and protein expression （respectively F = 4 083, 171, all P 〈 0. 001 ） were lower than those in control, the invasiveness （ respectively F = 125. 8, 48. 18, all P 〈 0. 001 ） was stronger than that in control. Tumor volume and weight in quereetin group, spermin group and control group were 2. 07 ±0. 90 vs. 6. 84 ±2. 15 vs. 7. 02 ＋2. 30. Pulmonary metastasis rate in three groups were 0/8 vs. 4/8 vs. 5/8, Nm23-H1 protein expression of quercetin and spermin group compared with control was upregulated or downregulated, respectively （P 〈 0.05 ）. Conclusion Quereetin can upregulate Nm23-H1 expression in hepatocellular carcinoma, and suppress the invasiveness and metastasis of hepatocellular carcinoma.