Effect Of Puerarin on The Growth of Nasopharyngeal Carcinoma Cells and Its Impact on Angiogenesis

Ye Li, Jingjing Zhao, Bo Yang
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Abstract

Objective Puerarin is a form of isoflavones obtained from Pueraria lobata. It stimulates hepatic metabolic function and lowers serum ALT, AST, and total-bilirubin level. The purpose of this study was to examine the effect of puerarin on nasopharyngeal carcinoma (NPC) CNE1 cells and preliminarily explore its possible mechanism. Materials and Methods CCK8 method was used to detect the proliferation activity of puerarin on NPC CNE1 cells and IC50 was calculated. CNE1 cells were treated with 0 μmol/L puerarin (containing equal volume of DMSO solution) as control group and 1000 μmol/L puerarin (IC50 concentration) as experimental group. Colony formation assay, Scratch-wound test and Transwell invasion assay were used to detect the clone formation ability, migration and invasion ability of puerarin on CNE1 cells. Then, RNA Sequencing was used to detect the changes of differentially expressed genes (DEGs) and signaling pathways after puerarin was applied to CNE1 cells. Results The inhibitory effect of puerarin on the proliferation activity of CNE1 cells was enhanced with the increase of concentration, and IC50 was calculated as 1000 μmol/L. Compared with the control group, the treatment of CNE1 cells with 1000 μmol/L puerarin could inhibit the clone formation, migration and invasion of CNE1 cells (P<0.05). A total of 379 DEGs were found by RNA sequencing, including 295 down-regulated genes and 84 up-regulated genes (padj<0.05). The significant differences in biological functions of differentially expressed genes were mainly distributed in “negative regulation of growth”, “angiogenesis”, “regulation of peptidase activity”, “positive regulation of vasculature development”, “digestion”, “positive regulation of angiogenesis”, “negative regulation of peptidase activity”, “extracellular matrix” and “Golgi lumen” (padj<0.05). Conclusion Puerarin could inhibit the proliferation, migration and invasion of NPC CNE1 cells, and its mechanism might be related to the inhibition of angiogenesis and cell growth.
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葛根素对鼻咽癌细胞生长及血管生成的影响
目的葛根素是从葛根中提取的一种异黄酮。它刺激肝脏代谢功能,降低血清ALT、AST和总胆红素水平。本研究旨在观察葛根素对鼻咽癌CNE1细胞的影响,并初步探讨其可能的作用机制。材料与方法采用CCK8法检测葛根素对鼻咽癌CNE1细胞的增殖活性,计算IC50。以0μmol/L葛根素(含等体积二甲基亚砜溶液)作为对照组,以1000μmol/L葛根素(IC50浓度)作为实验组。采用集落形成试验、划痕试验和Transwell侵袭试验检测葛根素对CNE1细胞的克隆形成能力、迁移和侵袭能力。然后,利用RNA测序技术检测葛根素应用于CNE1细胞后差异表达基因(DEGs)和信号通路的变化。结果葛根素对CNE1细胞增殖的抑制作用随浓度的增加而增强,IC50为1000μmol/L。与对照组相比,1000μmol/L葛根素对CNE1细胞的克隆形成、迁移和侵袭均有抑制作用(P<0.05),其中下调基因295个,上调基因84个(padj<0.05)。差异表达基因生物学功能的显著差异主要分布在“生长负调控”、“血管生成”、“肽酶活性调控”、,结论葛根素能抑制鼻咽癌CNE1细胞的增殖、迁移和侵袭,其作用机制可能与抑制血管生成和细胞生长有关。
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