Pu Yanchun, Jin Ping, Liu Lianghong, Pu Qinlin, Wu Fangping
School of Pharmaceutical Sciences, Hunan University of Medicine, No. 492, Jinxi South Road, Huaihua, Hunan Province 418099, China.
Evid Based Complement Alternat Med. 2021 Oct 20;2021:1221899. doi: 10.1155/2021/1221899. eCollection 2021.
In this study, we aim to investigate the effect of extract on biological behavior of esophageal cancer cells and its underlying mechanisms.
A total of 30 BALB/C nude mice (class SPF) were equally and randomly divided into the control group, model group, and group. CP-C cell of esophageal cancer was subcutaneously injected into the model group as well as the group, and the same amount of normal saline into the control group, in order to compare the tumorigenesis of nude mice of three groups. Wnt, -catenin, and p-GSK3/GSK3 expression in tumor tissues was detected using Western blot. CP-C cells in logarithmic growth were selected and divided into 4 groups, including the control group, podophyllotoxin group, Wnt activator group, and combined group (mixture of podophyllotoxin and Wnt activator). The cell viability, apoptosis, and invasion ability, Wnt, -catenin, and p-GSK3/GSK3 expression level of CP-C cells in each group were detected via MTT assay, flow cytometry, transwell, and Western blot, respectively.
The tumorigenesis rates of the control group, model group, and group were 0%, 90% (1 tumor-free mouse), and 80% (2 tumor-free mice), respectively. The tumor mass in the group was significant less than that in the model group. Based on the results of Western blot, Wnt, -catenin, and p-GSK3/GSK3 expression of the group was lower than that of the control group. The in vitro viability test indicated that there was a significant difference in cell viability exhibited among four groups. Cell viability level in the 3 groups, including the combined group, blank group, and Wnt activator group, was higher than the podophyllotoxin group at each time point. In vitro apoptosis assay revealed that significant differences in cell apoptosis exhibited among four groups. Cell apoptosis rate was higher in the podophyllotoxin group compared to the remaining three groups. The Wnt activator group showed the lowest cell apoptosis rate. The in vitro invasion assay demonstrated that numbers of transmembrane cell in the 3 groups, involving the combined group, blank group, and Wnt activator group, showed a higher level than the podophyllotoxin group. The results of Western blot manifested that the podophyllotoxin group showed lower level of Wnt, -catenin, and p-GSK3/GSK3 expression compared to the other 3 groups.
Podophyllotoxin in has an excellent antiesophageal cancer effect and is able to inhibit cell viability as well as invasion ability and promote apoptosis of esophageal cancer cells by inhibiting the Wnt signaling pathway, which could be potentially used in future clinical treatment of esophageal cancer.
在本研究中，我们旨在探讨提取物对食管癌细胞生物学行为的影响及其潜在机制。
将30只BALB/C裸鼠（SPF级）平均随机分为对照组、模型组和[提取物组（此处原文缺失提取物组的具体名称）]组。将食管癌CP-C细胞皮下注射到模型组以及[提取物组（此处原文缺失提取物组的具体名称）]组，对照组注射等量生理盐水，以比较三组裸鼠的肿瘤发生情况。采用蛋白质免疫印迹法检测肿瘤组织中Wnt、β-连环蛋白和p-GSK3/GSK3的表达。选取对数生长期的CP-C细胞，分为4组，包括对照组、鬼臼毒素组、Wnt激活剂组和联合组（鬼臼毒素与Wnt激活剂混合物）。分别通过MTT法、流式细胞术、Transwell实验和蛋白质免疫印迹法检测每组CP-C细胞的活力、凋亡、侵袭能力以及Wnt、β-连环蛋白和p-GSK3/GSK3的表达水平。
对照组、模型组和[提取物组（此处原文缺失提取物组的具体名称）]组的肿瘤发生率分别为0%、90%（1只无瘤小鼠）和80%（2只无瘤小鼠）。[提取物组（此处原文缺失提取物组的具体名称）]组的肿瘤块明显小于模型组。基于蛋白质免疫印迹法结果，[提取物组（此处原文缺失提取物组的具体名称）]组的Wnt、β-连环蛋白和p-GSK3/GSK3表达低于对照组。体外活力测试表明，四组细胞活力存在显著差异。联合组、空白组和Wnt激活剂组在各时间点的细胞活力水平均高于鬼臼毒素组。体外凋亡实验显示，四组细胞凋亡存在显著差异。鬼臼毒素组的细胞凋亡率高于其余三组。Wnt激活剂组的细胞凋亡率最低。体外侵袭实验表明，联合组、空白组和Wnt激活剂组的穿膜细胞数高于鬼臼毒素组。蛋白质免疫印迹法结果显示，鬼臼毒素组的Wnt、β-连环蛋白和p-GSK3/GSK3表达水平低于其他三组。
[提取物（此处原文缺失提取物的具体名称）]中的鬼臼毒素具有良好的抗食管癌作用，能够通过抑制Wnt信号通路抑制食管癌细胞的活力和侵袭能力，促进其凋亡，有望用于未来食管癌的临床治疗。
