Effect and mechanism of Xuanfu Daizhe Decoction on proliferation, migration and invasion activities of esophageal cancer cells
-
摘要:目的
探讨旋覆代赭汤对食管癌细胞增殖、迁移和侵袭活性的影响及机制。
方法使用不同质量浓度的旋覆代赭汤处理人食管癌细胞系EC109, 并依据旋覆代赭汤的质量浓度将细胞分为高浓度组(200 μg/mL)、中浓度组(100 μg/mL)、低浓度组(50 μg/mL)和空白组(0 μg/mL)。分别采用CCK-8实验、划痕实验和Transwell侵袭实验检测各组细胞的增殖、迁移和侵袭活性。采用蛋白质印迹法(Western blot)和细胞免疫荧光染色法检测各组细胞中糖酵解相关酶[己糖激酶2(HK2)、乳酸脱氢酶A(LDHA)、磷酸果糖激酶1(PFK1)]的蛋白表达水平, 并采用实时荧光定量聚合酶链式反应(qRT-PCR)检测上述酶编码基因mRNA相对表达量。构建裸鼠负瘤模型，分为对照组和旋覆代赭汤饲喂组(20 mg/kg), 观察旋覆代赭汤对食管癌体内生长的影响。
结果CCK-8实验、划痕实验和Transwell侵袭实验结果显示，旋覆代赭汤可浓度依赖性抑制EC109细胞的增殖、迁移和侵袭活性。Western blot和细胞免疫荧光分析结果显示，与空白组相比，低浓度组、中浓度组、高浓度组细胞的HK2、LDHA、PFK1蛋白表达水平降低，差异有统计学意义(P < 0.05); qRT-PCR检测结果显示，与空白组相比，低浓度组、中浓度组、高浓度组细胞的HK2 mRNA、LDHA mRNA、PFK1 mRNA相对表达量均降低，差异有统计学意义(P < 0.01或P < 0.000 1)。覆代赭汤饲喂组裸鼠背部皮下的肿瘤体积小于对照组，肿瘤组织中LDHA蛋白表达水平低于对照组，促凋亡蛋白Bax表达水平高于对照组，差异有统计学意义(P < 0.05)。
结论旋覆代赭汤可浓度依赖性抑制食管癌细胞糖酵解过程，进而抑制细胞增殖、迁移、侵袭及体内肿瘤生长。
Abstract:ObjectiveTo investigate the effect and mechanism of Xuanfu Daizhe Decoction on the proliferation, migration, and invasion activities of esophageal cancer cells.
MethodsHuman esophageal cancer cell line EC109 was treated with Xuanfu Daizhe Decoction at different concentrations, and the cells were divided into high-concentration group (200 μg/mL), medium-concentration group (100 μg/mL), low-concentration group (50 μg/mL), and blank group (0 μg/mL) based on the concentration. CCK-8 assay, wound healing assay, and Transwell invasion assay were used to detect the proliferation, migration, and invasion activities of the cells in each group, respectively. Western blot and cellular immunofluorescence staining were employed to detect the protein expression levels of glycolysis-related enzymes [hexokinase 2 (HK2), lactate dehydrogenase A (LDHA), and phosphofructokinase 1 (PFK1)] in the cells of each group, and real-time quantitative fluorescent polymerase chain reaction (qRT-PCR) was used to detect the relative expression levels of mRNA encoding these enzymes. A nude mouse tumor-bearing model was established and divided into control group and Xuanfu Daizhe Decoction-fed group (20 mg/kg) to observe the effect of Xuanfu Daizhe Decoction on the growth of esophageal cancer in vivo.
ResultsThe results of CCK-8 assay, wound healing assay, and Transwell invasion assay showed that Xuanfu Daizhe Decoction could inhibit the proliferation, migration, and invasion activities of EC109 cells in a concentration-dependent manner. Western blot and cellular immunofluorescence analysis revealed that compared with the blank group, the protein expression levels of HK2, LDHA, and PFK1 in the low-concentration, medium-concentration, and high-concentration groups were decreased (P < 0.05). The qRT-PCR results showed that compared with the blank group, the relative expression levels of HK2 mRNA, LDHA mRNA, and PFK1 mRNA in the low-concentration, medium-concentration, and high-concentration groups were all reduced(P < 0.01 or P < 0.000 1). The tumor volume in the subcutaneous tissue on the back of nude mice in the Xuanfu Daizhe Decoction-fed group was smaller than that in the control group, and the protein expression level of LDHA in the tumor tissue of the Xuanfu Daizhe Decoction-fed group was lower than that in the control group, while the expression level of the pro-apoptotic protein Bax was higher than that in the control group (P < 0.05).
ConclusionXuanfu Daizhe Decoction can inhibit the glycolysis process of esophageal cancer cells in a concentration-dependent manner, thereby inhibiting cell proliferation, migration, invasion, and tumor growth in vivo.
-
Keywords:
- Xuanfu Daizhe Decoction /
- esophageal cancer /
- glycolysis /
- proliferation /
- migration /
- invasion
-
-
![]()
图 1 不同质量浓度旋覆代赭汤对EC109细胞增殖、迁移和侵袭活性的影响
A: 各组EC109细胞增殖活性的CCK-8实验检测结果; B: 各组EC109细胞迁移活性的划痕实验检测结果; C: 各组EC109细胞侵袭活性的Transwell侵袭实验检测结果。与空白组比较, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.000 1。
![]()
图 2 不同质量浓度旋覆代赭汤处理对食管癌细胞糖酵解的影响
A: 各组EC109细胞糖酵解相关酶蛋白表达水平的Western blot检测结果; B: 各组EC109细胞糖酵解相关酶蛋白表达水平的细胞免疫荧光检测结果; C: 各组EC109细胞糖酵解相关酶编码基因mRNA相对表达量的qRT-PCR检测结果(与空白组比较, **P < 0.01, ****P < 0.000 1)。
![]()
图 3 旋覆代赭汤饲喂对负瘤模型裸鼠肿瘤生长的影响
A、B: 2组负瘤模型裸鼠肿瘤生长情况比较; C: 2组裸鼠肿瘤组织中糖酵解和凋亡相关蛋白表达水平的Western blot检测结果。
表 1 引物序列
基因 引物序列(5′→3′) GAPDH 正向: TGTGGGCATCAATGGATTTGG 反向: ACACCATGTATTCCGGGTCAAT HK2 正向: AATGGACAACTGGTCGTGGAC 反向: CCCTCCAGGGGATCTGTTTG LDHA 正向: CTCACCGGATGCACCAATGTT 反向: CGCGTTGCTCACAATGTTCAT PFK1 正向: GGAGCGAGATCCCTCCAAAAT 反向: GGCTGTTGTCATACTTCTCATGG 
下载: 导出CSV
-
[1] BÖHME F, RACZ K, SEBESTA C J, et al. Das ösophaguskarzinom[J]. Wien Med Wochenschr, 2023, 173(9): 209-215.
[2] ZHU H, MA X, YE T, et al. Esophageal cancer in China: Practice and research in the new era[J]. Int J Cancer, 2023, 152(9): 1741-1751. doi: 10.1002/ijc.34301
[3] LUENGO A, GUI D Y, VANDER HEIDEN M G. Targeting metabolism for cancer therapy[J]. Cell Chem Biol, 2017, 24(9): 1161-1180. doi: 10.1016/j.chembiol.2017.08.028
[4] BARBA I, CARRILLO-BOSCH L, SEOANE J. Targeting the Warburg effect in cancer: where do we stand[J]. Int J Mol Sci, 2024, 25(6): 3142. doi: 10.3390/ijms25063142
[5] CHELAKKOT C, CHELAKKOT V S, SHIN Y, et al. Modulating glycolysis to improve cancer therapy[J]. Int J Mol Sci, 2023, 24(3): 2606. doi: 10.3390/ijms24032606
[6] ZHENG S, LI H, LI Y, et al. The emerging role of glycolysis and immune evasion in gastric cancer[J]. Cancer Cell Int, 2023, 23(1): 317. doi: 10.1186/s12935-023-03169-1
[7] 翟冬妍, 吕冠华, 范颖. 旋覆代赭汤现代临床应用概述[J]. 中医药临床杂志, 2024, 36(2): 382-386. [8] 田晶晶, 袁红霞, 张月林, 等. 基于Caspase-3/Bcl-2/Bax信号通路探究加味旋覆代赭汤治疗食管癌前病变的作用机制[J]. 中国中西医结合外科杂志, 2024, 30(2): 258-264. doi: 10.3969/j.issn.1007-6948.2024.02.022 [9] 马媛, 荀敬, 王波涛, 等. 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. [10] 姚一菲, 孙可欣, 郑荣寿. 《2022全球癌症统计报告》解读: 中国与全球对比[J]. 中国普外基础与临床杂志, 2024, 31(7): 769-780. [11] TAKEUCHI H, MIYATA H, GOTOH M, et al. A risk model for esophagectomy using data of 5354 patients included in a Japanese nationwide web-based database[J]. Ann Surg, 2014, 260(2): 259-266. doi: 10.1097/SLA.0000000000000644
[12] ELLIOTT J A, DOCHERTY N G, ECKHARDT H G, et al. Weight loss, satiety, and the postprandial gut hormone response after esophagectomy: a prospective study[J]. Ann Surg, 2017, 266(1): 82-90. doi: 10.1097/SLA.0000000000001918
[13] 孙千惠, 朱广辉, 许博文, 等. 基于气陷理论探析恶性肿瘤术后中医药治疗与康复思路[J]. 中医杂志, 2024, 65(11): 1120-1125. [14] 李奕, 庞博, 花宝金, 等. 胃肠恶性肿瘤从"通道"立论的中医药治疗策略与思考[J]. 中华中医药杂志, 2023, 38(11): 5344-5347. [15] 田硕, 刘万里, 苏坤涵, 等. 旋覆代赭汤临床应用研究进展[J]. 亚太传统医药, 2021, 17(7): 205-209. [16] 邹利艳. 旋覆代赭汤联合针刺治疗胃癌术后胃瘫综合征31例[J]. 湖南中医杂志, 2020, 36(4): 52-54. [17] 向梅, 赵东丽, 梁宪斌, 等. 旋覆代赭汤联合耳穴按压对黑色素瘤化疗后不良反应的临床观察[J]. 实用中医内科杂志, 2020, 34(5): 109-112. [18] 谭林深, 朴瑛, 郑振东. 旋覆代赭汤加味治疗胰腺癌术后外分泌功能不全临床疗效观察[J]. 临床军医杂志, 2020, 48(1): 44-46. -
期刊类型引用(6)
1. 孟凯. 电视胸腔镜辅助环抱器治疗胸部外伤合并肋骨骨折患者的临床效果观察. 大医生. 2025(04): 23-25 . 
百度学术
2. 王晓康. 肋骨接骨板置入内固定术对肋骨骨折安全性及疼痛的影响. 中国医药指南. 2024(04): 14-16 . 百度学术
3. 成刚，王长兴. 基于胸腔镜的复位内固定术治疗老年肋骨骨折患者的临床疗效研究. 实用临床医药杂志. 2024(05): 89-93 . 本站查看
4. 彭亮，郭剑，查波辉，吴珊燕，舒华宝，吴胜华，杨子熠，王小娟. 急诊创伤性血气胸在床边超声引导下精准化穿刺置管引流术的治疗效果. 检验医学与临床. 2024(09): 1245-1249 . 百度学术
5. 冉隆强，李勇，贾萌. 胸腔镜辅助内固定术对多发肋骨骨折合并胸部创伤患者肺功能及预后的影响. 中国医刊. 2023(05): 517-520 . 百度学术
6. 周攀，吴中权，朱杰，舒圣，程玲玲. 胸腔镜微创手术和快速康复治疗严重胸部碾轧伤1例. 中国医药. 2023(08): 1255-1258 . 百度学术
其他类型引用(0)
计量
- 文章访问数: 54
- HTML全文浏览量: 10
- PDF下载量: 7
- 被引次数: 6
苏公网安备 32100302010246号