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Archive > Volume 30 Issue 4 > 2023,30(4):318-323. DOI:10.3872/j.issn.1007-385X.2023.04.006 Prev Next
Basic Research
Neobavaisoflavone induces pyroptosis in hepatocellular carcinoma Huh-7 cells through caspase-3/GSDME signal pathway
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Abstract:
Objective: To explore the effect of neobavaisoflavone (NBIF) on pyroptosis of hepatocellular carcinoma (HCC) Huh-7 cells and its molecular mechanism. Methods: Huh-7 cells were cultured in vitro, and the effects of different concentrations of NBIF on cell survival rate was detected by CCK-8. Morphological changes of Huh-7 cells treated with NBIF were observed under optical microscope.Lactate dehydrogenase (LDH) release test was used to detect the LDH release of cells. The protein levels of gasdermin E (GSDME) and caspase-3 in the cells were detected by WB assay. After silencing caspase-3 and GSDME in Huh-7 cells, the effect of NBIF treatment on cell survival rate was detected by CCK-8. The expression level of GSDME protein was detected by WB assay. The effect of NBIF treatment on cell morphology was observed, and the release of LDH was detected. Results: NBIF above 60 μmol/L could significantly inhibit the proliferation of Huh-7 cells (all P<0.01). The swelling and bubbling phenomenon of NBIF treated cells were observed under optical microscope, and the release of LDH was increased (all P<0.01). WB results showed that NBIF could activate caspase-3 protein, cleave GSDME protein and increase the expression of GSDME-N (P<0.01). After silencing caspase-3 and GSDME genes, the inhibition of NBIF on cells was weakened (all P<0.01), and the expression of GSDME-N protein was inhibited (P<0.01). Under the microscope, the phenomenon of cell swelling and bubbling almost disappeared, and the release of LDH decreased significantly (P<0.05). Conclusion: NBIF can cause pyroptosis of Huh-7 cells through caspase3-GSDME pathway, thus inhibiting the proliferation of Huh-7 cells, which provides a new idea for the treatment of HCC.
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