Calycosin induces autophagy and apoptosis via Sestrin2/AMPK/mTOR in human papillary thyroid cancer cells

Abstract

Calycosin, one of small molecules derived from astragalus, has anti-tumor effects in various tumors. However, the effects of calycosin on papillary thyroid cancer (PTC) remain unclear. This study aimed to explore the anti-tumor ability of calycosin on human PTC and its potential mechanisms. The B-CPAP cells were treated with calycosin, then cell proliferation, apoptosis and invasiveness were measured by CCK8 assay, flow cytometry, wound healing and transwell invasion assay, respectively. The cells were also performed by whole transcriptome microarray bioinformatics analysis. Apoptosis and autophagy-related markers or proteins were measured by qRT-PCR or western blot. Sestrin2-mediated AMPK/mTOR pathways were determined by western blot. We found that calycosin inhibited migration and invasion of B-CPAP cells and induced apoptosis (Bax/Bcl-2) and autophagy (LC3II/I, Beclin1) of B-CPAP cells. Differential expressed genes were screened between the calycosin-treated cells and control (524 genes upregulated and 328 genes downregulated). The pathway enrichment suggested that the role of calycosin in B-CPAP cells is closely related to apoptosis-related genes and p70S6 Kinase. Transmission electron microscopy found an increase in autophagosomes in calycosin-treated cells. Sestrin2 in human PTC tissues and B-CPAP cells was lower than in normal thyroid tissues and cells. And the pharmacological effects of calycosin in PTC cells were related to Sestrin2 activation, increased p-AMPK and inhibited p-mTOR and p-p70S6Kinase; these alterations were reversed when silencing Sestrin2. In conclusion, calycosin has an inhibitory effect on PTC via promoting apoptosis and autophagy through the Sestrin2/AMPK/mTOR pathway.

Unveiling the potential anti‐cancer activity of calycosin against multivarious cancers with molecular insights: A promising frontier in cancer research

“…Malignant cell formation in the tissue of the thyroid gland results in the most common endocrine tumor, that is, thyroid cancer. Calycosin has an inverse effect on thyroid cancer, for example, anti‐inflammatory, antioxidant, and anti‐tumor 66 . Human papillary thyroid cancer cells (B‐CPAP) and human normal thyroid cells (Nthy‐ori 3‐1) were treated with calycosin (0–100 μM), and the result demonstrated that calycosin suppresses the proliferation of cancer cells (B‐CPAP) 66 .…”

“…66 The general pharmacological effects on human papillary thyroid cancer cells that are related to the activation of Sestrin2 (SESN2), upregulated p-AMPK (phosphorylated AMPK), and inhibits p-mTOR (phosphorylated mTOR) and p-p70S6Kinase (ribosomal protein S6 kinase). 66 Overall, calycosin induced anti-tumor, antiinflammatory, and antioxidant function and decreased cancer cell proliferation while regulating many pathways in thyroid cancer therapy.…”

“…Calycosin has an inverse effect on thyroid cancer, for example, anti‐inflammatory, antioxidant, and anti‐tumor 66 . Human papillary thyroid cancer cells (B‐CPAP) and human normal thyroid cells (Nthy‐ori 3‐1) were treated with calycosin (0–100 μM), and the result demonstrated that calycosin suppresses the proliferation of cancer cells (B‐CPAP) 66 . The general pharmacological effects on human papillary thyroid cancer cells that are related to the activation of Sestrin2 (SESN2), upregulated p‐AMPK (phosphorylated AMPK), and inhibits p‐mTOR (phosphorylated mTOR) and p‐p70S6Kinase (ribosomal protein S6 kinase) 66 …”

“…Human papillary thyroid cancer cells (B‐CPAP) and human normal thyroid cells (Nthy‐ori 3‐1) were treated with calycosin (0–100 μM), and the result demonstrated that calycosin suppresses the proliferation of cancer cells (B‐CPAP) 66 . The general pharmacological effects on human papillary thyroid cancer cells that are related to the activation of Sestrin2 (SESN2), upregulated p‐AMPK (phosphorylated AMPK), and inhibits p‐mTOR (phosphorylated mTOR) and p‐p70S6Kinase (ribosomal protein S6 kinase) 66 …”

Exploring the multi-targeting phytoestrogen potential of Calycosin for cancer treatment: A review

“…[3,4] Additionally, it has been shown to offer protective effects on the heart, blood vessels, nerves, liver, kidney and against cancer. [5][6][7][8][9] Notably, several studies have highlighted the potential of CA as an anti-metastatic agent in various tumors, promoting apoptosis in cancer cells while exhibiting low toxicity to normal cells. [10] Despite these promising findings, the specific mechanism of action and its impact on different cancers remains poorly understood.…”