Page 64 - 2020年2月第31卷第3期

瓜蒂水提物和醇提物对食管癌 TE-1、EC-1 细胞的增殖、迁移、克

隆形成的影响及机制研究 Δ

＊
赵雯宇 ，司富春（河南中医药大学河南省中医方证信号传导重点实验室/河南省中医方证信号传导国际联合实
#
验室，郑州 450046）

中图分类号 R735.1；R965 文献标志码 A 文章编号 1001-0408（2020）03-0314-07
DOI 10.6039/j.issn.1001-0408.2020.03.12

摘 要 目的：研究瓜蒂水提物和醇提物对食管癌 TE-1、EC-1 细胞增殖、迁移、克隆形成的影响及作用机制。方法：体外培养
TE-1、EC-1细胞，分别加入质量浓度均为0、1.562 5、3.125、6.25、12.5、25、50、100、200 μg/mL的瓜蒂水提物或醇提物（以提取物粉
末计）进行培养，采用 MTT 法检测细胞的生长抑制率并计算半数抑制浓度（IC50 ）。将 TE-1、EC-1 细胞分为 TE-1/EC-1 空白组、
TE-1/EC-1瓜蒂水提物组（药液浓度均为IC50 ）、TE-1/EC-1瓜蒂醇提物组（药液浓度均为IC50 ），采用实时无标记细胞分析（RTCA）法
检测各组细胞的增殖与迁移，绘制细胞增殖、迁移曲线；采用显微镜观察各组细胞形态学的变化；采用软琼脂克隆形成试验分析各
组细胞克隆形成能力的变化，并计算细胞克隆形成率；采用流式细胞仪检测各组细胞的细胞周期、凋亡率；采用Western blotting检
测各组细胞表皮生长因子受体（EGFR）、蛋白激酶C-α（PKC-α）的相对表达量。结果：瓜蒂水提物作用于TE-1、EC-1细胞的IC50分
别为49.24、76.38 μg/mL，瓜蒂醇提物作用于TE-1、EC-1细胞IC50分别为9.08、14.53 μg/mL；瓜蒂水提物和醇提物加药后30 h内对
细胞有增殖抑制作用；瓜蒂水提物和醇提物加药后60 h内对细胞有迁移抑制作用。与TE-1/EC-1空白组比较，各加药组细胞数量
明显减少，细胞结构松散，多数细胞轮廓消失、变圆等；细胞克隆形成率显著下降（P＜0.01）；G2期细胞百分率显著升高（P＜0.01），
G1期、S 期细胞百分率显著降低（P＜0.05）；早、晚期凋亡率均显著增加（P＜0.05）；EGFR、PKC-α蛋白相对表达量显著降低（P＜
0.01）。与TE-1/EC-1瓜蒂水提物组比较，TE-1/EC-1瓜蒂醇提物组细胞克隆形成率显著下降（P＜0.05）；G2期细胞率显著升高（P＜
0.05）；EGFR、PKC-α蛋白相对表达量显著降低（P＜0.01）；TE-1瓜蒂醇提物组早、晚期细胞凋亡率显著降低（P＜0.05）；EC-1瓜蒂
醇提物组早、晚期细胞凋亡率显著升高（P＜0.05）。结论：瓜蒂水提物和醇提物可影响TE-1、EC-1细胞增殖、迁移、克隆形成的能
力，促进细胞凋亡，其作用机制可能与下调EGFR、PKC-α蛋白有关。
关键词 瓜蒂；食管癌；水提物；醇提物；细胞增殖；细胞迁移；克隆形成；作用机制

Effects and Mechanism of Water Extract and Ethanol Extract of Muskmelon Pedicel on the Proliferation，
Migration and Cloning Formation of Esophageal Carcinoma TE-1 and EC-1 Cells
ZHAO Wenyu，SI Fuchun（Provincial Key Laboratory of Prescription-Syndrome Signal Transduction of Chinese
Medicine & International Joint Laboratory of Prescription-Syndrome Signal Transduction of Chinese Medicine in
Henan Province，Henan University of Chinese Medicine，Zhengzhou 450046，China）

ABSTRACT OBJECTIVE：To study the effects and mechanism of water extract and ethanol extract of Muskmelon Pedicel on the
proliferation，migration and cloning formation of esophageal carcinoma TE-1 and EC-1 cells. METHODS：TE-1 and EC-1 cells
were cultured in vitro，and were treated with 0，1.562 5，3.125，6.25，12.5，25，50，100，200 μg/mL of water extract and ethanol
extract of Muskmelon Pedicel（calulated by extract powder）. MTT assay was used to detect the growth inhibitory rate of TE-1 and
EC-1 cells，and calculate IC50 of them. TE-1 and EC-1 cells were divided into TE-1/EC-1 blank group，TE-1/EC-1 Muskmelon
Pedicel water extract group（IC50 as drug concentration），and TE-1/EC-1 Muskmelon Pedicel ethanol extract group（IC50 as drug
concentration）. The proliferation and migration of cells in each group were detected by real-time unlabeled cell analysis（RTCA），
and cell proliferation and migration curves were drawn. The morphological changes of cells were observed under microscope；soft
agarose colony forming test was used to analyze the change of colony forming ability of cells in each group，and the colony
forming rate was calculated；cell cycle and apoptosis rate of cells in each group were detected by flow cytometry；Western blotting
assay was used to detect the relative expression of EGFR and PKC-α in cells in each group. RESULTS：IC50 of water extract of
Muskmelon Pedicel were 49.24，76.38 μg/mL respectively for TE-1 and EC-1 cells. Those of ethanol extract of Muskmelon Pedicel
were 9.08，14.53 μ g/mL respectively for TE-1 and EC-1 cells. The inhibition effect of water extract and ethanol extract of
Muskmelon Pedicel on the cell proliferation were within 30 h.
Δ 基金项目：河南省自然科学基金资助项目（No.162300410185）
The inhibition effect of water extract and ethanol extract of
＊博士研究生。研究方向：肿瘤中医方证。电话：0371-65676778。
E-mail：zixiangning88@126.com Muskmelon Pedicel on the cell migration were within 60 h.
# 通信作者：教授，博士生导师，博士。研究方向：肿瘤中医方 Compared with TE-1/EC-1 blank group，the number of cells
证。电话：0371-65676778。E-mail：sifc2000@hotmail.com was decreased significantly in administration groups， the

·314 · China Pharmacy 2020 Vol. 31 No. 3 中国药房 2020年第31卷第3期