Professor Wang Shaokang, Southeast University, et al.: Inhibition effect and mechanism of forsythiaside on esophageal epithelial malignant transformation cells and esophageal cancer Eca-109 cells

According to statistics, the incidence and mortality of esophageal cancer in China remain high, and it is a high-incidence malignant tumor
that threatens national health.
At present, the treatment of esophageal cancer includes surgical resection, chemotherapy and radiotherapy, but the side effects cannot be ignored
.
Traditional Chinese medicine has a long history of application in China, rich resources, low cost, less toxic side effects, etc.
, so it is particularly important
to screen anti-cancer bioactive ingredients in traditional Chinese medicine and explore their anti-cancer mechanism.

A growing body of research has shown that forsythoside works through a variety of signaling pathways such as phosphatidylinositol-3-kinase/protein kinase B (PI3K/PKB) (also known as PI3K/AKT), Janus kinase/signal transduction and transcriptional activator (JAK/STAT), p38 mitogen-activated protein kinase (p38 MAPK), and more
。 Guo Ziqi and Wang Shaokang* from the School of Public Health, Southeast University, took esophageal epithelial malignant transformation cells (self-made by this group) and Eca-109 cells as the research objects to observe the effects of forsythoside on the proliferation, cycle, apoptosis, migration and invasion of these two cells, and explored its mechanism of action from the PI3K/AKT signaling pathway, which provided a scientific basis
for the further development and utilization of forsythia leaf, a new food raw material.

1.
Determination of esophageal epithelial malignant transformation cells

As shown in Fig.
1, the cloning formation rates of HEEC and esophageal epithelial malignant transformation cells were (0.
60±0.
08)% and (4.
23±0.
21)%, respectively, and compared with HEEC, the cloning formation rate of esophageal epithelial malignant transformation cells was significantly higher (P<0.
05), indicating that the cell proliferation ability of esophageal epithelial malignant transformation cells was stronger<b10>.
Studies have shown that the precancerous lesions of esophageal squamous cell carcinoma mainly refer to the dysplasia of esophageal squamous epithelial cells, so it is believed that the malignant transformation cells of the esophageal epithelium have a certain tumorigenic ability and belong to the esophageal precancerous cells
.

2.
Effects of forsythoside on the survival rate of esophageal epithelial malignant transformation cells, Eca-109 cells and HEEC

The results of CCK8 method (Table 1) showed that 10, 100, 200, 400, 600, 800 μmol/L and 1 000 μmol/L forsythoside could inhibit the proliferation of esophageal epithelial malignant transformation cells and Eca-109 cells, and their IC50 values were 308.
4 μmol/L and 322.
0 μmol/L, respectively.
However, 1, 10, and 100 μmol/L forsythoside promoted HEEC proliferation, and 200, 400, 600, 800 μmol/L, and 1 000 μmol/L forsythoside inhibited HEEC proliferation with IC50 values of 384.
4 μmol/L
。 Combined with the effect of forsythoside on the proliferation of three kinds of cells, 1, 10 and 100 μmol/L forsythoside had no obvious inhibition of HEEC proliferation, which proved that forsythoside concentration had no obvious cytotoxic effect on HEEC when it was lower than or equal to 100 μmol/L.
However, 10, 100 μmol/L forsythia glycoside can inhibit the proliferation of esophageal epithelial malignant transformation cells and Eca-109 cells, and in order to avoid the effect of high concentration of forsythianoside cytotoxicity, the concentration of 10 and 100 μmol/L was selected for follow-up experiments
.

3.
Inhibition effect and mechanism of forsythoside on malignant transformation cells of esophageal epithelium

Effect of forsythoside on the cell cycle of malignant transition in the esophageal epithelium

The results of flow cytometry cell cycle detection (Figure 2) showed that compared with the 0 μmol/L intervention group, the cell cycle of malignant transformation in the esophageal epithelium of the esophageal epithelium in the 10 and 100 μmol/L forsythoside intervention groups was significantly changed, the proportion of cells in the G0/G1 phase in the intervention group increased significantly (P<0.
05), and the proportion of cells in the S phase was significantly decreased (P<0.
05), and with the increase of forsythoside concentration, the more obvious the change in the cell ratio of each cycle, indicating that forsythoside may be blocked by blocking cells in G0/ The G1 phase inhibits the proliferation<b10> of esophageal epithelial malignant transforming cells.

Effect of forsythiaside on apoptosis of esophageal epithelial malignant transformation cells