Page 35 - 《中国药房》2021年13期

阿魏酸对人肝癌HepG2细胞增殖、侵袭和凋亡的影响 Δ

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音金萍 ，卓少元（广西中医药大学基础医学院，南宁 530200）
中图分类号 R965 文献标志码 A 文章编号 1001-0408（2021）13-1565-07
DOI 10.6039/j.issn.1001-0408.2021.13.05

摘 要 目的：研究阿魏酸对人肝癌 HepG2 细胞增殖、侵袭和凋亡的影响。方法：采用 CCK-8 法筛选阿魏酸的给药浓度；采用
Western blot法筛选白细胞介素6（IL-6）诱导磷酸化信号转导蛋白及转录激活子3（p-STAT3）蛋白高表达的HepG2细胞模型的最佳
造模浓度。将HepG2细胞分为空白对照组、模型组、阿魏酸组（0.5 mmol/L）和阳性对照组（p-STAT3抑制剂C188-9，10 μmol/L），除
空白对照组外，模型组加入IL-6、各给药组加入IL-6和相应药物分别进行培养。采用CCK-8法、Transwell法和Annexin V-FITC/PI
双染色法分别检测各组细胞存活率、侵袭数和早期、晚期凋亡率；采用Western blot法检测各组细胞中p-STAT3、胱天蛋白酶3（cas-
pase-3）、锌指蛋白（ZBP-89）和波形蛋白（vimentin）的表达水平。基于PDB蛋白数据库，以STAT3高度相似晶体结构1BG1为对接
模板，以Tyr705周围的区域为假定的结合口袋，进行阿魏酸与STAT3蛋白的分子对接分析。结果：选择以0.5 mmol/L 阿魏酸干预
HepG2细胞48 h作为后续实验条件，以50 ng/mL IL-6为造模条件。与空白对照组比较，模型组细胞侵袭数和p-STAT3/STAT3比
值、vimentin 蛋白表达水平均显著增加或升高（P＜0.05 或 P＜0.01），细胞晚期凋亡率和 caspase-3 蛋白表达水平均显著降低（P＜
0.05或P＜0.01）。与模型组比较，阿魏酸组和阳性对照组细胞存活率、侵袭数、p-STAT3/STAT3比值和vimentin蛋白表达水平均显
著减少或降低（P＜0.05或P＜0.01），细胞早期凋亡率（阿魏酸组除外）、晚期凋亡率和caspase-3、ZBP-89（阳性对照组除外）蛋白表
达水平均显著升高（P＜0.05或P＜0.01）。分子对接结果表明，阿魏酸的羧酸基团与Asn581、Lys591分别存在1.9Å、2.0Å的氢键作
用，结合能为－4.4 kcal/mol。结论：阿魏酸可能通过与STAT3磷酸化位点直接结合来抑制p-STAT3的活性；并可通过STAT3依赖
性途径上调 caspase-3 蛋白表达，或可通过 STAT3 非依赖性途径上调 ZBP-89 蛋白表达，进而下调 vimentin 蛋白表达，从而抑制
HepG2细胞的增殖、侵袭与凋亡。
关键词 阿魏酸；人肝癌HepG2细胞；信号转导蛋白及转录激活子3；胱天蛋白酶3；波形蛋白

Effects of Ferulic Acid on the Proliferation，Invasion and Apoptosis of HepG2 Hepatocellular Carcinoma
Cells
YIN Jinping，ZHUO Shaoyuan（School of Basic Medical Science，Guangxi University of TCM，Nanning
530200，China）

ABSTRACT OBJECTIVE： To study the effects of ferulic acid on the proliferation， invasion and apoptosis of HepG2
hepatocelluar carcinoma cells. METHODS：CCK-8 assay was used to screen the concentration of ferulic acid. Western blot assay
was adopted to screen the optimal concentration of interleukin 6 （IL-6） to induce HepG2 cell model with high expression of
phosphorylated signal transduction protein and activator 3（p-STAT3）protein. HepG2 cell were divided into blank control group，
model group，ferulic acid group（0.5 mmol/L）and positive control group（p-STAT3 inhibitor C188-9，10 μmol/L）. Except for
blank control group，model group treated with IL-6，while administration groups were treated with IL-6 and relevant drugs. Cell
survival rate，invasion and apoptosis rate in early and late stage were detected by CCK-8 assay，Transwell assay and Annexin
V-FITC/PI double staining，respectively. Western blot assay was used to detect the expression of p-STAT3，caspase-3，ZBP-89 and
vimentin proteins in each group. On the basis of the PDB protein database，using 1BG1，a highly similar crystal structure of
STAT3，as docking template，using the region around Tyr705 as the putative binding pocket，the docking analysis of ferulic acid
with STAT3 protein was carried out. RESULTS：It is selected to use 0.5 mmol/L ferulic acid intervention for 48 h as the follow-up
experimental condition；50 ng/mL IL-6 was selected as the modeling condition. Compared with blank control group，the number of
cell invasion，p-STAT3/STAT3 ratio and protein expression of
Δ 基金项目：国家自然科学基金资助项目（No.81860824，No. vimentin were increased significantly in model group（P＜0.05
81660775）；2017 年广西中药药效研究重点实验室课题（No.17-259-
or P＜0.01），while late apoptosis rate and protein expression
20）；广西中医药大学研究生教育创新计划项目（No.YCSZ2020024）
of caspase-3 were decreased significantly （P＜0.05 or P＜
＊硕士研究生。研究方向：肝癌的中医药防治。E-mail：yinjin-
0.01）. Compared with model group，cell survival rate，the
ping2018@163.com
# 通信作者：教授，硕士生导师，博士。研究方向：肝癌的中医药 number of cell invasion，p-STAT3/STAT3 ratio and protein
防治。E-mail：sherwin_zhuo@126.com expression of vimentin were decreased significantly in ferulic

中国药房 2021年第32卷第13期 China Pharmacy 2021 Vol. 32 No. 13 ·1565 ·