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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 GreenNucTM 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.
CONCLUSIONS: Solasonine inhibits proliferation and induces apoptosis of PC9 cells by regulating the Bcl-2/Bax/caspase-3 pathway and its upstream proteins.
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 GreenNucTM 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.
CONCLUSIONS: Solasonine inhibits proliferation and induces apoptosis of PC9 cells by regulating the Bcl-2/Bax/caspase-3 pathway and its upstream proteins.
摘要：
目的：为了研究索拉索宁的作用，龙葵的活性成分，对非小细胞肺癌PC9细胞增殖和凋亡的影响。
方法：PC9细胞用2、5、10、15、20或25μmol/L索拉索宁处理，用CCK-8法检测细胞增殖的变化。四甲基罗丹明乙酯（TMRE）用于检测线粒体膜电位的变化，使用caspase-3/7检测试剂盒和GreenNucTMcaspase-3/AnnexinV-mCherry活细胞试剂盒分析细胞caspase-3的变化。采用膜联蛋白V-FITC/PI双染色分析细胞凋亡率。通过使用Western印迹检测凋亡相关蛋白的表达来检查PTEN抑制剂对索拉索宁诱导的细胞凋亡的影响。
结果：Solasonine处理24、48和72小时显着降低了PC9细胞的活力。用solasonine处理24h的细胞显示线粒体膜电位显着降低，细胞凋亡增加，caspase-3/7和caspase-3活性增强，caspase-3表达增强。索拉索宁处理显著降低PI3K和Akt的磷酸化水平，增加PTEN和Bax的蛋白表达，并降低细胞中Bcl-2蛋白的表达。
结论：索拉索宁可通过调控Bcl-2/Bax/caspase-3通路及其上游蛋白抑制PC9细胞增殖并诱导其凋亡。
方法：PC9细胞用2、5、10、15、20或25μmol/L索拉索宁处理，用CCK-8法检测细胞增殖的变化。四甲基罗丹明乙酯（TMRE）用于检测线粒体膜电位的变化，使用caspase-3/7检测试剂盒和GreenNucTMcaspase-3/AnnexinV-mCherry活细胞试剂盒分析细胞caspase-3的变化。采用膜联蛋白V-FITC/PI双染色分析细胞凋亡率。通过使用Western印迹检测凋亡相关蛋白的表达来检查PTEN抑制剂对索拉索宁诱导的细胞凋亡的影响。
结果：Solasonine处理24、48和72小时显着降低了PC9细胞的活力。用solasonine处理24h的细胞显示线粒体膜电位显着降低，细胞凋亡增加，caspase-3/7和caspase-3活性增强，caspase-3表达增强。索拉索宁处理显著降低PI3K和Akt的磷酸化水平，增加PTEN和Bax的蛋白表达，并降低细胞中Bcl-2蛋白的表达。
结论：索拉索宁可通过调控Bcl-2/Bax/caspase-3通路及其上游蛋白抑制PC9细胞增殖并诱导其凋亡。
