Kaempferol suppresses glioma progression and synergistically enhances the antitumor activity of gefitinib by inhibiting the EGFR/SRC/STAT3 signaling pathway

by Selleckchem | Oct 20 2023

Kaempferol (Kae) is a natural flavonoid that has multiple biological activities, such as anti-inflammatory and antitumor activities. However, few studies have been reported on antiglioma effects of Kae. This study aimed to explore the effects and potential mechanisms of Kae and synergistic antitumor activities with gefitinib (Gef) on glioma. Cell Counting Kit-8 and 5-ethynyl-2'-deoxyuridine assays were used to detect cytotoxicity and cell proliferation. Cell apoptosis and the cell cycle were detected by flow cytometry. Transwell assays were used to detect the migratory and invasive abilities of glioma cells. Network pharmacology and molecular docking analysis were used to screen for core targets of Kae in glioma therapy. Xenograft tumor nude mice were established with U251 cells to verify the antiglioma effects of Kae in vivo. A terminal deoxynucleotidyl transferase dUTP nick end labeling assay was used to detect apoptosis in tumor tissues. The expression of proteins was detected by immunohistochemistry and western blot analysis. Kae inhibited cell proliferation, promoted apoptosis, and induced cell cycle arrest in the G2/M phase of glioma cells in a concentration-dependent manner. Kae inhibited the migration and invasion of glioma cells at low concentrations. Network pharmacology analyses showed that epidermal growth factor receptor (EGFR) and SRC proto-oncogene (SRC) might be direct molecular-binding targets of Kae. Our results showed that Kae inhibited the levels of phosphorylated EGFR, phosphorylated SRC (p-SRC), and phosphorylated signal transducer and activator of transcription 3 (STAT3). In addition, the combination of Kae with Gef significantly inhibited the proliferation of glioma cells. Kae further inhibited EGFR phosphorylation after treatment with Gef. Similarly, Kae further enhanced the inhibition of p-SRC caused by SU6656. Finally, we demonstrated that Kae exerted great antitumor activities and enhanced the antitumor effect of Gef by inhibiting EGFR/SRC/STAT3 signaling pathway in vivo. Kae played a potential role and synergistic antiglioma effects with Gef by inhibiting the phosphorylation of EGFR/SRC dual targets. Kae is expected to be a candidate drug or chemosensitizer in glioma therapy.

Comments:

The study you mentioned aimed to investigate the effects and potential mechanisms of kaempferol (Kae), a natural flavonoid, on glioma, as well as its synergistic antitumor activities with gefitinib (Gef), a drug used in cancer treatment. Here is a breakdown of the key findings and methods used in the study:

1. Cell Counting Kit-8 and 5-ethynyl-2'-deoxyuridine assays were conducted to assess the cytotoxicity and cell proliferation, respectively. The results demonstrated that Kae inhibited cell proliferation in glioma cells in a concentration-dependent manner.

2. Flow cytometry analysis revealed that Kae promoted apoptosis (programmed cell death) and induced cell cycle arrest in the G2/M phase of glioma cells.

3. Transwell assays were employed to evaluate the migratory and invasive abilities of glioma cells. The study found that Kae inhibited the migration and invasion of glioma cells, particularly at low concentrations.

4. Network pharmacology and molecular docking analysis were utilized to identify the core targets of Kae in glioma therapy. The analysis suggested that epidermal growth factor receptor (EGFR) and SRC proto-oncogene (SRC) may be direct molecular-binding targets of Kae.

5. In vivo experiments using xenograft tumor nude mice established with U251 cells were conducted to verify the antiglioma effects of Kae. A terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was performed to detect apoptosis in the tumor tissues.

6. Immunohistochemistry and western blot analysis were employed to assess the expression of proteins. The study found that Kae inhibited the phosphorylation (activation) of EGFR, SRC, and signal transducer and activator of transcription 3 (STAT3), which are involved in various cellular processes.

7. The study investigated the synergistic antitumor effects of Kae and Gef. The combination of Kae and Gef significantly inhibited the proliferation of glioma cells. Kae further inhibited EGFR phosphorylation when combined with Gef. Similarly, Kae enhanced the inhibition of p-SRC caused by SU6656, a SRC kinase inhibitor.

8. The study concluded that Kae exhibited potent antiglioma activities and enhanced the antitumor effect of Gef by inhibiting the EGFR/SRC/STAT3 signaling pathway. Kae was proposed as a potential candidate drug or chemosensitizer in glioma therapy.

In summary, this study demonstrated that kaempferol (Kae) exerted multiple effects on glioma cells, including inhibiting cell proliferation, promoting apoptosis, and inhibiting migration and invasion. The study also identified EGFR and SRC as potential molecular targets of Kae. Furthermore, the combination of Kae with Gef exhibited synergistic antitumor effects by targeting the EGFR/SRC/STAT3 signaling pathway. These findings suggest that Kae may have therapeutic potential for glioma treatment and could be further explored as a drug candidate or chemosensitizer.