Effect of Shidao Tongjie Formula on esophageal cancer cell‑derived exosomal LncRNA ZFAS1 induced angiogenesis in human vascular endothelial cells

CAI Xiaoyue; WANG Hui; ZHOU Yaning; GUO Yijun; ZHANG Zhouji; ZHANG Ming

doi:10.16305/j.1007-1334.2026.z20250606001

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Effect of Shidao Tongjie Formula on esophageal cancer cell‑derived exosomal LncRNA ZFAS1 induced angiogenesis in human vascular endothelial cells

更新时间：2026-02-09

- Effect of Shidao Tongjie Formula on esophageal cancer cell‑derived exosomal LncRNA ZFAS1 induced angiogenesis in human vascular endothelial cells
- Shanghai Journal of Traditional Chinese Medicine Vol. 60, Issue 2, Pages: 77-85(2026)
- 作者机构：
  
  1.上海交通大学医学院附属胸科医院中西医结合科（上海 200030）
  2.上海交通大学医学院研究生院（上海 200025）
- 作者简介：
- 基金信息：
- DOI：10.16305/j.1007-1334.2026.z20250606001
  CLC：
- Received：06 June 2025，
  
  Published：10 February 2026
- 稿件说明：
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蔡霄月,王慧,周亚宁,等.食道通结方对食管癌来源外泌体LncRNA ZFAS1诱导人血管内皮细胞血管生成的影响[J].上海中医药杂志,2026,60(2):77-85.

CAI Xiaoyue,WANG Hui,ZHOU Yaning,et al.Effect of Shidao Tongjie Formula on esophageal cancer cell‑derived exosomal LncRNA ZFAS1 induced angiogenesis in human vascular endothelial cells[J].Shanghai Journal of Traditional Chinese Medicine,2026,60(2):77-85.
蔡霄月,王慧,周亚宁,等.食道通结方对食管癌来源外泌体LncRNA ZFAS1诱导人血管内皮细胞血管生成的影响[J].上海中医药杂志,2026,60(2):77-85. DOI： 10.16305/j.1007-1334.2026.z20250606001.

CAI Xiaoyue,WANG Hui,ZHOU Yaning,et al.Effect of Shidao Tongjie Formula on esophageal cancer cell‑derived exosomal LncRNA ZFAS1 induced angiogenesis in human vascular endothelial cells[J].Shanghai Journal of Traditional Chinese Medicine,2026,60(2):77-85. DOI： 10.16305/j.1007-1334.2026.z20250606001.

摘要

目的

探讨食道通结方对人食管癌Eca-109细胞来源外泌体诱导人脐静脉内皮细胞（HUVECs）血管生成的影响。

方法

20只雄性SD大鼠，随机分为食道通结方含药血清组和空白鼠血清组，每组10只，分别以51.3 g/kg食道通结方及等体积0.9%氯化钠溶液灌胃，制备含药血清。分别以20%无外泌体空白血清、20%食道通结方含药血清（体积分数）处理Eca-109细胞；以超速离心法分别提取Eca-109细胞来源外泌体（Eca-109外泌体），以及经食道通结方含药血清处理的Eca-109细胞来源外泌体（食道通结方外泌体）；采用电镜、纳米颗粒跟踪分析（NTA）法、Western blot法鉴定外泌体，实时荧光定量逆转录聚合酶链式反应（RT-qPCR）检测外泌体长链非编码RNA锌指结构反义转录本1（LncRNA ZFAS1）的表达。将外泌体与HUVECs共培养，免疫荧光染色法观察外泌体被HUVECs摄取的情况。5-乙炔基-2'-脱氧尿苷（EdU）细胞增殖、血管形成实验观察外泌体对HUVECs血管生成的影响，RT-qPCR检测HUVECs中LncRNA ZFAS1

、

血管内皮生长因子A（

VEGFA

）mRNA的表达，Western blot法检测HUVECs中VEGFA蛋白的表达。

结果

电镜、NTA及Western blot法检测分化簇63（CD63）、肿瘤易感基因101蛋白（TSG101）、钙联蛋白（Calnexin）的结果表明，两组提取物均鉴定为

外泌体。荧光显微镜下观察到两组外泌体均被HUVECs摄取。食道通结方可降低Eca-109细胞来源外泌体LncRNA ZFAS1的表达（

0.01）。与空白对照组比较，Eca-109细胞外泌体能诱导HUVECs的增殖、显著增加成管（

0.05，

0.01），LncRNA ZFAS1

、VEGFA

mRNA及VEGFA蛋白的表达显著上调（

0.05）；而食道通结方能显著降低Eca-109细胞外泌体诱导HUVECs增殖、成管的能力（

0.05，

0.01），使HUVECs中LncRNA ZFAS1

、VEGFA

mRNA及VEGFA蛋白的表达显著下调（

0.05）。

结论

Eca-109细胞来源外泌体LncRNA ZFAS1可诱导HUVECs血管形成，食道通结方可能通过降低Eca-109细胞来源外泌体LncRNA ZFAS1的表达，下调HUVECs中LncRNA ZFAS1、VEGFA的表达，抑制HUVECs增殖、成管的能力，从而起到拮抗食管癌肿瘤血管新生的作用。

Abstract

Objective

To explore the effect of Shidao Tongjie Formula on angiogenesis of HUVECs induced by exosomes derived from Eca‑109 esophageal cancer cells.

Methods

Twenty male SD rats were randomly divided into the Shidao Tongjie Formula drug serum group and the blank serum group， with 10 rats in each group. Two groups of rats were respectively administered 51.3 g/kg of the Shidao Tongjie Formula and the same volume of 0.9% sodium chloride solution by gavage to prepare the drug serum. Eca-109 cells were treated with 20% exosome-free blank serum or 20% Shidao Tongjie Formula drug serum （volume fraction）. The exosomes derived from Eca-109 cells （Eca-109 exosomes） as well as the exosomes derived from Eca-109 cells that had been treated with the Shidao Tongjie Formula drug serum （ Shidao Tongjie Formula exosomes） were isolated by ultracentrifugation method. The exosomes were identified by electron microscopy， nanoparticle tracking analysis （NTA） and Western blot. Real-time fluorescence quantitative reverse transcription polymerase chain reaction （RT-qPCR） was used to detect the expression of long non-coding RNA zinc finger structure antisense transcript 1 （LncRNA ZFAS1） in exosomes. Exosomes were co-cultured with HUVECs， and the uptake of exosomes by HUVECs was

observed using immunofluorescence staining. The effect on the angiogenesis of HUVECs was evaluated by 5-ethynyl- 2'-deoxyuridine （EdU） and tube formation assays. LncRNA ZFAS1 and

Vascular endothelial growth factor A （

VEGFA

） mRNA in HUVECs were measured by RT-qPCR， and VEGFA protein in HUVECs was determined by Western blot.

Results

The results of electron microscopy， NTA and Western blot for the marker proteins of differentiation cluster 63 （CD63）， tumor susceptibility gene 101 （TSG101）， and calnexin indicated that both groups of extracts were identified as exosomes. It was observed that both groups of exosomes were taken up by HUVECs under fluorescence microscope. The expression of LncRNA ZFAS1 in Eca-109 cell derived exosomes was significantly decreased by Shidao Tongjie Formula （

0.01）. Compared with the control group， the proliferation and tube formation of HUVECs induced by Eca-109 cells derived exosomes were significantly promoted （

0.05，

0.01）， and the expressions of LncRNA ZFAS1，

VEGFA

mRNA and VEGFA protein in HUVECs cells were significantly upregulated （

0.05）. While Shidao Tongjie Formula could significantly reduce the ability of Eca‑109 cells derived exosomes to induce the proliferation and tube formation of HUVECs （

0.05，

0.01）， and significantly downregulate the expressions of LncRNA ZFAS1，

VEGFA

mRNA and VEGFA protein in HUVECs cells （

0.05）.

Conclusions

Eca-109 cells derived exosomal LncRNA ZFAS1 can promote angiogenesis in HUVECs. Shidao Tongjie Formula may inhibit esophageal cancer angiogenesis by reducing Eca-109 cells derived exosomal LncRNA ZFAS1 expression， downregulating LncRNA ZFAS1 and VEGFA expression in HUVECs， and suppressing HUVECs proliferation and tube formation.

关键词

Keywords

references

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