Solasonine promotes apoptosis of non-small cell lung cancer cells by regulating the Bcl-2/Bax/caspase-3 pathway

doi:10.12122/j.issn.1673-4254.2024.06.11

Abstract

Abstract:

Objective To investigate the effect of solasonine, an active component of Solanum nigrum, on proliferation and apoptosis of non-small cell lung cancer PC9 cells. Methods PC9 cells were treated with 2, 5, 10, 15, 20, or 25 μmol/L solasonine, and the changes in cell proliferation were examined using CCK-8 assay. Tetramethyl rhodamine ethyl ester (TMRE) was used to detect the changes in mitochondrial membrane potential, and caspase-3/7 detection kit and GreenNuc™ caspase-3/Annexin V-mCherry kit for live cell were used to analyze the changes in caspase-3 of the cells. Annexin V-FITC/PI double staining was employed to analyze the apoptosis rate of the cells. The effect of PTEN inhibitors on solasonine-induced cell apoptosis was examined by detecting apoptosis-related protein expressions using Western blotting. Results Solasonine treatment for 24, 48, and 72 h significantly lowered the viability of PC9 cells. The cells treated with solasonine for 24 h showed significantly decreased mitochondrial membrane potential and increased cell apoptosis with enhanced caspase-3/7 and caspase-3 activities and expression of cleaved caspase-3. Solasonine treatment significantly decreased phosphorylation levels of PI3K and Akt, increased the protein expressions of PTEN and Bax, and lowered the expression of Bcl-2 protein in the cells. Conclusion Solasonine inhibits proliferation and induces apoptosis of PC9 cells by regulating the Bcl-2/Bax/caspase-3 pathway and its upstream proteins.

Key words: solasonine, lung cancer, Bcl-2/Bax/caspase-3 signaling pathway, homologous phosphatase tensin protein, apoptosis

Guiling CHEN, Xiaofeng LIAO, Pengtao SUN, Huan CEN, Shengchun SHU, Bijing LI, Jinhua LI. Solasonine promotes apoptosis of non-small cell lung cancer cells by regulating the Bcl-2/Bax/caspase-3 pathway[J]. Journal of Southern Medical University, 2024, 44(6): 1109-1116.

Figures/Tables 7

Fig.1 Effect of solasonine on PC9 cell proliferation (Mean±SD, n=3). *P<0.01, **P<0.001 vs Control.

Fig.2 Changes of mitochondrial membrane potential in solasonine-treated PC9 cells detected using TMRE and observed by immunofluorescence microscopy.

Fig.4 Western blotting for detecting the expression of cleaved caspase-3 in PC9 cells treated with 5, 10, 15, 20 and 25 μmol/L solasonine for 24 h and quantitative analysis of the protein expression levels using Image J (n=3). A: Protein bands in Western blotting. B, C: Histogram of the protein expression levels. **P<0.01 vs Control.

Fig.5 Activity of caspase-3 in PC9 cells detected using GreenNuc™ caspase-3/Annexin V-mCherry double staining and observed by immunofluorescence microscopy.

Fig.6 Western blotting for detecting expressions of Bcl-1 and Bax in PC9 cells treated with 5, 10, 15, 20 and 25 μmol/L solasonine for 24 h and quantitative analysis of the protein expression levels (n=3). *P<0.05, **P<0.01 vs Control.

Fig.7 Western blotting for detecting the expressions of p-PI3K and p-Akt in PC9 cells treated with 20 μmol/L solasonine l for 2, 4, 12 and 24 h and quantitative analysis of the protein expression levels (n=3). *P<0.05, **P<0.01 vs Control.

Fig.8 Effect of solasonine on protein expression of PTEN in PC9 cells. n=3, *P<0.05, **P<0.01 vs Control, ▲P<0.05 vs SF1670.

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