[1]金 秋,刘永芳,李志国,等.莱菔素调控ANO1/LXR通路降低食管癌细胞胆固醇代谢并抑制侵袭迁移的实验研究[J].现代检验医学杂志,2026,41(01):11-14+57.[doi:10.3969/j.issn.1671-7414.2026.01.003]
　JIN Qiu,LIU Yongfang,LI Zhiguo,et al.Experimental Study of Sulforaphene Regulation of the ANO1 / LXR Pathway to Reduce Cholesterol Metabolism and Inhibit Invasion and Migration in Esophageal Cancer Cells[J].Journal of Modern Laboratory Medicine,2026,41(01):11-14+57.[doi:10.3969/j.issn.1671-7414.2026.01.003]
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莱菔素调控ANO1/LXR通路降低食管癌细胞胆固醇代谢并抑制侵袭迁移的实验研究()

《现代检验医学杂志》[ISSN:/CN:]

卷:
第41卷
期数:
2026年01期
页码:
11-14+57
栏目:
论著
出版日期:
2026-01-15

文章信息/Info

Title:
Experimental Study of Sulforaphene Regulation of the ANO1 / LXR Pathway to Reduce Cholesterol Metabolism and Inhibit Invasion and Migration in Esophageal Cancer Cells
文章编号:
1671-7414（2026）01-011-05
作者:
金 秋1a，刘永芳1a，李志国2，范花平2，孙鹏博2，程丽敏1b
1. 邯郸市中心医院 a.内科；b.消化内科，河北邯郸 056001；2. 武安市人民医院胸外科，河北武安 056300
Author(s):
JIN Qiu1a，LIU Yongfang1a，LI Zhiguo2，FAN Huaping2，SUN Pengbo2，CHENG Limin1b
1a. Department of Internal Medicine; 1b. Department of Gastroenterology, Handan Central Hospital, Hebei Handan 056001, China；2. Department of Thoracic Surgery, Wu’an People’s Hospital, Hebei Wu’an 056300, China
关键词:
食管癌; 莱菔素; 钙激活氯通道蛋白1（ANO1）/肝X受体（LXR）通路; 胆固醇代谢物; 侵袭; 迁移
分类号:
R735.1；R730.4
DOI:
10.3969/j.issn.1671-7414.2026.01.003
文献标志码:
A
摘要:
目的?探究莱菔素调控钙激活氯通道蛋白1（ANO1）/肝X受体（LXR）通路对食管癌细胞侵袭迁移及胆固醇代谢物水平的影响及可能机制。方法培养人正常食管细胞（HEEC）、人食管癌细胞EC109、KYSE510、KYSE150、KYSE30及TE-11，实时荧光定量PCR（RT-qPCR）检测细胞中ANO1与LXR的mRNA水平，筛选出EC109细胞为后期研究对象。使用不同浓度莱菔素干预EC109细胞，CCK-8试剂盒检测EC109细胞活性，筛选出20μmol/L莱菔素为后续研究浓度。继续培养EC109细胞，分组为食管癌组、莱菔素组及过表达ANO1组。Transwell实验、划痕实验检测EC109细胞侵袭、迁移能力；蛋白免疫印迹（Westernblotting）检测EC109细胞中ANO1与LXR的蛋白水平；酶联免疫吸附试验（ELISA）检测EC109细胞中胆固醇含量；免疫荧光染色检测EC109细胞中代谢相关蛋白ATP结合盒转运体-1（ABCA1）、ATP枸橼酸裂解酶（ACLY）、肽酰脯氨酰异构酶B（PPIB）蛋白水平。结果与HEEC细胞相比，食管癌细胞EC109、KYSE510、KYSE150、KYSE30及TE-11中ANO1mRNA表达增加，LXRmRNA表达减少，差异具有统计学意义（F=90.77、59.76，均P<0.05），其中EC109细胞的ANO1与LXR表达最为显著，作为后期研究对象。使用不同浓度莱菔素（0、5、10、20、40、80μmol/L）处理食管癌细胞EC10948h，随着莱菔素浓度的增加，食管癌细胞活性逐渐降低，差异具有统计学意义（F=454.80，P<0.05），其中20μmol/L约为半数细胞活性抑制浓度，因此选该浓度继续后续实验。与食管癌组相比，莱菔素组食管癌细胞的侵袭及迁移能力降低，ANO1蛋白表达减少，LXR蛋白表达增加，胆固醇含量及代谢相关蛋白ABCA1、ACLY、PPIB蛋白表达减少，差异具有统计学意义（t=7.22～17.70，均P<0.05）；与莱菔素组相比，过表达ANO1组食管癌细胞ANO1蛋白表达增加，LXR蛋白表达减少，胆固醇含量及代谢相关蛋白ABCA1、ACLY、PPIB蛋白表达增加，差异具有统计学意义（t=6.11～10.87，均P<0.05）。结论莱菔素可调控ANO1/LXR通路抑制食管癌细胞侵袭迁移能力，这可能与该通路调控胆固醇代谢物水平相关。
Abstract:
Objective To investigate the effects of sulforaphene on invasion, metastasis and cholesterol metabolite levels of esopha-geal cancer cells, and its potential mechanisms through regulating the calcium-activated chloride channel protein 1 (ANO1)/liver X receptor (LXR) pathway. Methods Human esophageal epithelial cells (HEEC) and esophageal cancer cell lines EC109, KYSE510, KYSE150, KYSE30 and TE-11 were cultured. Real-time quantitative PCR (RT-qPCR) was used to detect mRNA levels of ANO1 and LXR in the cells. EC109 cells were selected as the objects for subsequent studies. EC109 cells were treated with different con-centrations of sulforaphene, and cell viability was assessed using CCK-8 kit, and 20 μmol/L sulforaphene was selected as the con-centration for subsequent studies. EC109 cells were further cultured and divided into esophageal cancer control group, sulforaphene group and ANO1 overexpression group. The invasive and migration ability of EC109 cells was detected by Transwell assays and scratch assays. Protein levels of ANO1 and LXR in EC109 cells were detected by Western blotting. Cholesterol content in EC109 cells was detected by ELISA. The levels of metabolism-related proteins ATP-binding cassette transporter-1 (ABCA1), ATP citrate lyase (ACLY) and peptidylprolyl isomerase B (PPIB) in EC109 cells were detected by immunofluorescence staining. Results Compared with HEEC cells, esophageal cancer cells EC109, KYSE510, KYSE150, KYSE30 and TE-11 cells exhibited increased ANO1 mRNA level and decreased LXR mRNA level, with statistically significant differences (F=90.77, 59.76, all P<0.05). Among these cell lines, EC109 showed the most pronounced ANO1 and LXR expressions, making it the subject of subsequent studies. Treatment of EC109 cells with different concentrations of sulforaphene (0, 5, 10, 20, 40, 80 μmol/L) for 48h rendered the progres-sive decrease of cell viability with increasing sulforaphene concentration (F=454.80, P<0.05). 20 μmol/L of sulforaphene inhibited approximately half of the cell viability, thus this concentration was selected for subsequent experiments. Compared with esophageal cancer control group, sulforaphene group exhibited reduced invasion and migration capacity, decreased ANO1 protein expression, increased LXR protein expression , and reduced cholesterol content along with decreased expression of cholesterol metabolism-re-lated proteins ABCA1, ACLY and PPIB (t=7.22 ~ 17.70, all P<0.05). Compared with sulforaphene group, the ANO1-overexpressing group exhibited increased ANO1 protein expression, decreased LXR protein expression (t=10.61, 8.48, P<0.001), increased choles-terol content, and elevated expression of cholesterol metabolism-related proteins ABCA1, ACLY, PPIB (t=6.11 ~ 10.87, all P<0.05). Conclusions Sulforaphene can regulate the ANO1/LXR pathway to inhibit the invasion and metastasis of esophageal cancer cells, which may be related to its regulation of cholesterol metabolite levels.

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备注/Memo

备注/Memo:
基金项目：河北省卫健委2023年度医学科学研究课题（20231946）。
作者简介：金秋（1987-），女，本科，主治医师，研究方向：食管癌，E-mail：jvsnoo86@163.com。
通讯作者：程丽敏（1985-），女，硕士，副主任医师，研究方向：食管癌。
更新日期/Last Update: 2026-01-15