Effect of Xuanfu Daizhe decoction on stemness of esophageal cancer cells

doi:10.12047/j.cjap.6228.2022.012

Abstract

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

CLC Number:

R2-031

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.

References

[1] Smyth EC, Lagergren J, Fitzgerald RC, et al. Oesophageal cancer[J]. Nat Rev Dis Primers, 2017, 3(1): 17048.
[2] Clarke MF, Dick JE, Dirks PB, et al. Cancer stem cells--perspectives on current status and future directions: AACR Workshop on cancer stem cells[J]. Cancer Res, 2006, 66(19): 9339-9344.
[3] 蔡昂, 向国辉, 杨全伟, 等. 张德忠运用旋覆代赭汤加减治疗上消化道肿瘤经验[J]. 中医肿瘤学杂志, 2020, 2(5): 63-66.
[4] 刘亚婷, 刘菊, 苗嘉萌, 等. 旋覆代赭汤对反流性食管炎大鼠模型TLR4/NF-κB的影响[J]. 中国中西医结合杂志, 2020, 40(1): 80-84.
[5] 黄棪, 徐明英, 夏雪皎, 等. 旋覆代赭汤对胆碱能抗炎通路相关RE大鼠炎症因子的影响[J]. 天津中医药大学学报, 2018, 37(5): 386-389.
[6] 唐丽明, 宋宁, 张桂贤, 等. 加味旋覆代赭汤对非酸反流所致慢性食管病变大鼠肠化及CDX2、DLK1表达的影响[J]. 中国中西医结合外科杂志, 2020, 26(1): 2-7.
[7] 陈丽娟, 王向群, 余杨桦, 等. 旋覆代赭汤治疗功能性消化不良及对胃肠激素影响的临床观察[J]. 中华中医药学刊, 2019, 37(2): 417-420.
[8] 杨梦婷, 高乐, 王相, 等. 经典名方旋覆代赭汤的研究进展[J]. 中药药理与临床, 2021, 37(3): 214-221.
[9] Pophali P, Halland M. Barrett's oesophagus: diagnosis and management [J]. BMJ, 2016, 353: i2373.
[10] Vaira D, Gatta L, Ricci C, et al. Gastroesophageal reflux disease and Barrett's esophagus[J]. Intern Emerg Med, 2011, 6(4): 299-306.
[11] 任吉莲, 崔灵芝, 郝小夏, 等. 厚朴酚联合吉非替尼对A549非小细胞肺癌细胞的干预作用[J]. 中国应用生理学杂志, 2021, 37(5): 506-510.
[12] 曹启迪, 逯艳艳, 马颖才. 食管癌的早诊早治 [J]. 临床消化病杂志, 2021, 33(2): 141-143.
[13] Short M, Burgers K, Fry V. Esophageal cancer [J]. Am Fam Physician, 2017, 95(1): 22-28.
[14] 文珍, 张彦秋, 吴蓉, 等. 首发为食管鳞癌的多原发癌患者临床特征及生存分析 [J]. 中国应用生理学杂志, 2021, 37(4): 407-414.
[15] 马长德, 张超云. 旋覆代赭汤加味治疗中晚期食管癌19例[J]. 中医临床研究, 2011, 3(19): 53-54.
[16] 黄兆迁. 旋覆代赭汤加减治疗食管癌38例的体会 [J]. 医学信息(中旬刊), 2011, 24(1): 334.
[17] Lu P, Liang QD, Li R, et al. Effect of traditional Chinese medicine on survival and quality of life in patients with esophageal carcinoma after esophagectomy[J]. Chin J Integr Med, 2006, 03): 175-179.
[18] 刘浩江. 旋覆代赭汤治疗食管癌手术后並发症46例[J]. 河南中医, 1987, 7(1): 22.
[19] 崔亚, 姜礼双, 卜平. 旋覆代赭汤实验研究及临床应用进展 [J]. 江苏中医药, 2018, 50(1): 82-85.
[20] 袁红霞, 田晶晶, 闫早兴. 中医药治疗反流性食管炎研究近况 [J]. 世界华人消化杂志, 2016, 24(32): 4323-4329.
[21] Wang K, Zhang L, He ZH, et al. A population-based survey of gastroesophageal reflux disease in a region with high prevalence of esophageal cancer in China[J]. Chin Med J (Engl), 2019, 132(13): 1516-1523.
[22] Uko N, Güner O, Matesic D, et al. Akt pathway inhibitors[J]. Curr Top Med Chem, 2020, 20(10): 883-900.
[23] 阿友, 孟祥宁, 刘改云, 等. EGF通过激活FAK-P13K/AKT途径促进肺癌细胞增殖[J]. 国际遗传学杂志, 2007(4): 241-243.
[24] Gao H, Wang H, Peng J. Hispidulin induces apoptosis through mitochondrial dysfunction and inhibition of P13k/Akt signalling pathway in HepG2 cancer cells[J]. Cell Biochem Biophys, 2014, 69(1): 27-34.
[25] Ma X, Bai Y. IGF-1 activates the P13K/AKT signaling pathway via upregulation of secretory clusterin[J]. Mol Med Rep, 2012, 6(6): 1433-1437.
[26] Wang Z, Kang L, Zhang H, et al. AKT drives SOX2 overexpression and cancer cell stemness in esophageal cancer by protecting SOX2 from UBR5-mediated degradation[J]. Oncogene, 2019, 38(26): 5250-5264.