旋覆代赭汤对食管癌细胞干性的影响*

doi:10.12047/j.cjap.6228.2022.012

中国应用生理学杂志 ›› 2022, Vol. 38 ›› Issue (1): 62-67.doi: 10.12047/j.cjap.6228.2022.012

旋覆代赭汤对食管癌细胞干性的影响^*

马媛¹, 荀敬², 王波涛¹, 李棣华², 张琦², 王震宇³, 吴瑜^4Δ

1.天津医科大学研究生院, 天津 300070;
2.天津医科大学南开临床学院, 天津市急腹症器官损伤与中西医修复重点实验室, 天津 300100;
3.天津医科大学南开临床学院, 肝胆胰外科三, 天津 300100;
4.天津医科大学南开临床学院, 胃肠外科三, 天津 300100

收稿日期:2021-11-28 修回日期:2022-01-26 出版日期:2022-01-28 发布日期:2022-05-30
通讯作者: ^△Tel: 15522320960; E-mail: wyctj3250@163.com
基金资助:
* 天津市中医药重点领域项目(2020009)

Effect of Xuanfu Daizhe decoction on stemness of esophageal cancer cells

MA Yuan¹, XUN Jing², WANG Bo-tao¹, LI Di-hua², ZHANG Qi², WANG Zhen-yu³, WU Yu^4Δ

1. Graduate School, Tianjin Medical University, Tianjin 300070;
2. Tianjin Key Laboratory of Acute Abdomen Disease Associated Organ Injury and ITCWM Repair, Nankai Clinical College, Tianjin Medical University, Tianjin 300100;
3. Department of Hepatobiliary Surgery, Nankai Clinical College, Tianjin Medical University, Tianjin 300100;
4. Department of Gastrointestinal Surgery, Nankai Clinical College, Tianjin Medical University, Tianjin 300100, China

Received:2021-11-28 Revised:2022-01-26 Online:2022-01-28 Published:2022-05-30

摘要/Abstract

摘要： 目的: 探究旋覆代赭汤对食管癌细胞干性的影响。方法: 将BALB/c裸鼠随机分为对照组与实验组,每组5只,分别连续给予生理盐水和旋覆代赭汤(9.89 g/kg)灌胃处理,在给予灌胃第8日时皮下接种5×10⁶细胞数量的人源食管癌ECA-109细胞,每周测量肿瘤大小,4周后处死小鼠。收取肿瘤组织和小鼠血清,采用RT-qPCR、Western blot和免疫组化方法检测肿瘤细胞干性相关转录因子NANOG、OCT4、SOX2的表达。分别用含10%胎牛血清和上述两组小鼠的血清处理48 h食管癌ECA-109细胞,每组设置三个复孔,用RT-qPCR和Western blot方法检测NANOG、OCT4、SOX2的mRNA和蛋白表达水平,以及AKT及其磷酸化(p-AKT)的蛋白表达水平;用流式细胞术检测肿瘤细胞中ALDH酶活性;用成球实验检测肿瘤细胞的成球数量。结果: 与对照组相比,旋覆代赭汤显著抑制食管癌小鼠肿瘤的生长和大小(P＜0.01或P＜0.05);旋覆代赭汤药物血清显著降低食管癌细胞的中NANOG、OCT4和SOX2 mRNA和蛋白的表达水平、ALDH酶活性和成球数量,以及AKT和AKT的磷酸化(p-AKT)水平(P＜0.01或P＜0.05)。结论: 旋覆代赭汤具有抑制食管癌细胞干性的作用,其可能是治疗食管癌的潜在有效药物,为食管癌治疗探寻新的有效药物提供理论依据。

关键词: 旋覆代赭汤, 食管癌, 肿瘤干细胞, 干性, 小鼠, 细胞培养

Abstract: Objective: To investigate the effect of Xuanfu Daizhe decoction on the stemness of esophageal cancer cells. Methods: The BALB/c nude mice were randomly divided into the control group and experimental group, 5 mice in each group, which were continuously administered with normal saline and Xuanfu Daizhe decoction (9.89 g/kg) by gastrogavage, respectively. Human esophageal carcinoma cells ECA-109 (5×10⁶) were subcutaneously injected into the mice on the 8th day. Tumor volume was measured twice a week. The mice were sacrificed 4 weeks after injection, and the tumor tissue and mouse serum were collected. The expressions of the major stemness-regulating transcription factors, i.e., NANOG, OCT4 and SOX2, were detected by RT-qPCR, Western Blot and immunohistochemistry. ECA-109 cells were treated with 10% fetal bovine serum and serum from the above two groups of mice for 48 hours respectively, and three replicate wells were set in each group, and the expressions of NANOG, OCT4, SOX2 and the levels of AKT and p-AKT were detected by RT-qPCR and Western Blot, respectively. ALDH activity in tumor cells was detected by flow cytometry; the number of spheroids of tumor cells was detected by the spheroidization experiment. Results: Compared with the control group, the growth and size of esophageal cancer tumors were significantly inhibited by Xuanfu Daizhe Decoction; the expressions of NANOG, OCT4, SOX2, the ALDH activity, the number of spheroids, and the levels of AKT and phosphorylated AKT (p-AKT) in esophageal cancer cells were significantly reduced by Xuanfu Daizhe Decoction both in vivo and in vitro. Conclusion: Xuanfu Daizhe Decoction inhibits the stemness of esophageal cancer cells, it may be a potentially effective drug for the treatment of esophageal cancer and provides a theoretical basis for the exploration of new effective drugs for the treatment of esophageal cancer.

Key words: Xuanfu Daizhe decoction, esophageal cancer, cancer stem cells, stemness of cancer cells, mice, cell culture

中图分类号:

R2-031

马媛, 荀敬, 王波涛, 李棣华, 张琦, 王震宇, 吴瑜. 旋覆代赭汤对食管癌细胞干性的影响^*[J]. 中国应用生理学杂志, 2022, 38(1): 62-67.

MA Yuan, XUN Jing, WANG Bo-tao, LI Di-hua, ZHANG Qi, WANG Zhen-yu, WU Yu. Effect of Xuanfu Daizhe decoction on stemness of esophageal cancer cells[J]. CJAP, 2022, 38(1): 62-67.

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旋覆代赭汤对食管癌细胞干性的影响^*

Effect of Xuanfu Daizhe decoction on stemness of esophageal cancer cells

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