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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
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| 篇名： | 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 |
| TITLE： | 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 |
| 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、1.5625、3.125、6.25、12.5、25、50、100、200μg/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Westernblotting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、76.38μg/mL，瓜蒂醇提物作用于TE-1、EC-1细胞IC50分别为9.08、14.53μg/mL；瓜蒂水提物和醇提物加药后30h内对细胞有增殖抑制作用；瓜蒂水提物和醇提物加药后60h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。 |
| 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 IC 50 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） ＊博士研究生。研究方向：肿瘤中医方证。电话：0371-65676778。 The inhibition effect of water extract and ethanol extract of 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 2020Vol. 31 No. 3 中国药房 2020年第31卷第3期 structure of cell were sloop ，the cell structure was loose ，and most of the cell contour disappeared and became round. The formation rate of cell clone was decreased significantly （P＜0.01）. The percentage of G 2 phase cells increased significantly （P＜ 0.01），while that of G 1 and S phase cells decreased significantly （P＜0.05）. The apoptotic rate of cells increased significantly in early and late stage （P＜0.05）. Relative protein expression of EGFR and PKC- α were decreased significantly （P＜0.01）. Compared with TE- 1/EC-1 Muskmelon Pedicel water extract group ，formation rate of cell clone was decreased significantly in TE- 1/EC-1 Muskmelon Pedicel ethanol extract group （P＜0.05）；cell was increased significantly at G 2 phase（P＜0.05）；relative protein expression of EGFR and PKC- α were decreased significantly （P＜0.01）. The apoptotic rate of cells in early and late stage in TE- 1 Muskmelon Pedicel ethanol extract group was decreased significantly （P＜0.05），the apoptotic rate of cells in early and late stage in EC- 1 Muskmelon Pedicel ethanol extract group was increased significantly （P＜0.05）. CONCLUSIONS ：Water extract and ethanol extract of Muskmelon Pedicel could influence the proliferation ，migration and clone formation ability of TE- 1 and EC- 1 cell，promote cell apoptosis ，the mechanism of which may be associated with the down-regulation of EGFR and PKC-α protein. |
| 期刊： | 2020年第31卷第03期 |
| 作者： | 赵雯宇，司富春 |
| AUTHORS： | ZHAO Wenyu ，SI Fuchun |
| 关键字： | 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 |
| KEYWORDS： | Muskmelon Pedicel ；Esophageal carcinoma ；Water extract ；Ethanol extract ；Cell proliferation ；Cell migration ； |
| 阅读数： | 284 次 |
| 本月下载数： | 9 次 |
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