绞股蓝水提物调控SRPX2/PI3K/Akt通路抑制食管癌细胞增殖、迁移和侵袭*

作者：张振悍1，郭建鑫1，张晨辰1，周 露1，李金歌1，崔子悦1，吴忠冰1，李 晶1,2

单位：1.河北医科大学中西医结合学院，河北 石家庄 050017； 2.河北医科大学第四医院，河北 石家庄 050019

引用：引用：张振悍，郭建鑫，张晨辰，周露，李金歌，崔子悦，吴忠冰，李晶.绞股蓝水提物调控SRPX2/PI3K/Akt通路抑制食管癌细胞增殖、迁移和侵袭[J].中医药导报,2026,32(3):6-11.

DOI：10.13862/j.cn43-1446/r.2026.03.002

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摘要：

目的：探究绞股蓝（GpM）水提物对食管癌细胞增殖、迁移和侵袭的影响及其作用机制。方法：制备不同浓度的GpM水提物药液，通过细胞计数试剂盒-8（CCK-8）实验检测GpM水提物对人源食管癌细胞KYSE-150、Eca-109和人永生化上皮细胞NE-1增殖能力的影响；通过细胞克隆形成实验检测GpM水提物对人源食管癌细胞克隆形成能力的影响；通过细胞划痕愈合实验检测GpM水提物对食管癌细胞迁移能力的影响；通过Transwell实验检测GpM水提物对食管癌细胞迁移和侵袭能力的影响；通过蛋白质印迹（Western blotting）法检测GpM水提物作用后各组SRPX2/PI3K/Akt信号通路关键蛋白（SRPX2、PI3K、p-PI3K、Akt、p-Akt）的表达。结果：CCK-8实验结果显示，GpM水提物可显著抑制食管癌细胞增殖[KYSE-150：48 h F（4，10）=49.96，P<0.001；3.2 mg/mL（42.63±5.20）%，P<0.001。Eca-109：48 h F（4，10）=28.53，P<0.001；3.2 mg/mL（38.27±6.70）%，P<0.001]，但对正常食管细胞NE-1活力的影响较弱[48 h 3.2 mg/mL处理组细胞活力保持（57.16±1.94）%]。细胞克隆形成实验证实，1.6 mg/mL GpM水提物可以显著抑制克隆形成[KYSE-150：F（2，6）=225.20，P<0.001；（87±2）个，P<0.001。Eca-109：F（2，6）=61.64，P<0.001；（106±25）个，P<0.001]。细胞划痕愈合实验表明，1.6 mg/mL GpM水提物能显著抑制细胞迁移[KYSE-150：F（2，6）=51.84，P<0.001；（14.96±0.31）%，P<0.001。Eca-109：F（2，6）=24.63，P=0.001；（7.33±3.51）%，P=0.001]。Transwell实验则进一步证实，该浓度GpM水提物既可抑制细胞迁移[KYSE-150：F（2，6）=15.31，P=0.004；（2 121±227）个，P=0.003]，也能抑制细胞侵袭[Eca-109：F（2，6）=6.02，P=0.037；（474±101）个，P=0.026]。Western blotting分析进一步揭示，1.6 mg/mL GpM水提物可有效下调SRPX2/PI3K/Akt信号通路活性[KYSE-150 SRPX2：F（2，6）=517.00，P<0.001；（0.06±0.03），P<0.001。Eca-109 p-Akt/Akt：F（2，6）=16.27，P=0.004；（0.63±0.08），P=0.002]。结论：GpM水提物能够抑制食管癌细胞的增殖、迁移和侵袭，并伴随SRPX2/PI3K/Akt通路关键蛋白表达的改变。

关键词：食管癌；绞股蓝；SRPX2/PI3K/Akt通路；细胞增殖；细胞迁移；细胞侵袭

Abstract：

Objective: To investigate the effect and mechanism of the water extract of Gynostemma pentaphyllum (Thunb.) Makino (GpM) on the proliferation, migration, and invasion of esophageal cancer cells. Methods: Different concentrations of GpM water extract were prepared. The Cell Counting Kit-8 (CCK-8) assay was used to detect the effect of GpM water extract on the proliferation of human esophageal cancer cell lines KYSE-150, Eca-109, and human immortalized esophageal epithelial cell line NE-1. The effect of GpM water extract on the colony formation ability of human esophageal cancer cells was detected by a colony formation assay. The effect of GpM water extract on the migration ability of esophageal cancer cells was detected by a wound healing assay. The effects of GpM water extract on the migration and invasion abilities of esophageal cancer cells were detected by Transwell assays. Western blotting was used to detect the expression of key proteins (SRPX2, PI3K, p-PI3K, Akt, p-Akt) in the SRPX2/PI3K/Akt signaling pathway in each group after treatment with GpM water extract. Results: CCK-8 assay results showed that the GpM water extract significantly inhibited the proliferation of esophageal cancer cells [KYSE-150: 48 h F(4,10)=49.96, P<0.001; at 3.2 mg/mL, cell viability was (42.63±5.20)%, P<0.001. Eca-109：48 h F(4,10)=28.53, P<0.001; at 3.2 mg/mL, cell viability was (38.27±6.70)%, P<0.001], but had a weaker effect on the viability of normal esophageal NE-1 cells [at 48 h, cell viability in the 3.2 mg/mL treatment group remained at (57.16±1.94)%]. The colony formation assay confirmed that 1.6 mg/mL GpM water extract significantly inhibited colony formation [KYSE-150：F(2,6)=225, P<0.001; colony number was (87±2), P<0.001. Eca-109：F(2,6)=61.64, P<0.001; colony number was (106±25), P<0.001]. The wound healing assay showed that 1.6 mg/mL GpM water extract significantly inhibited cell migration [KYSE-150：F(2,6)=51.84, P<0.001; migration rate was (14.96±0.31)%, P<0.001. Eca-109：F(2,6)=24.63, P=0.001; migration rate was (7.33±3.51)%, P=0.001]. The Transwell assay further confirmed that this concentration of GpM water extract inhibited both migration [KYSE-150：F(2,6)=15.31, P=0.004; migrated cell number was (2 121±227), P=0.003] and invasion [Eca-109：F(2,6)=6.02, P=0.037; invasive cell number was (474±101), P=0.026]. Western blotting analysis further revealed that 1.6 mg/mL GpM water extract effectively downregulated the activity of the SRPX2/PI3K/Akt signaling pathway (KYSE-150 SRPX2 protein expression: F(2,6)=517, P<0.001; expression level was (0.06±0.03), P<0.001. Eca-109 p-Akt/Akt ratio：F(2,6)=16.27, P=0.004; ratio was (0.63±0.08), P=0.002). Conclusion: GpM water extract can inhibit the proliferation, migration, and invasion of esophageal cancer cells, which is accompanied by changes in the expression of key proteins of the SRPX2/PI3K/Akt pathway.

Key words：esophageal cancer; Gynostemma pentaphyllum (Thunb.) Makino; SRPX2/PI3K/Akt pathway; cell proliferation; cell migration; cell invasion

发布时间：2026-03-28

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