DOI: 10.57760/sciencedb.j00217.05494
title: Study on Shengxian Tang-Containing serum inhibiting the malignant biological behavior of non-small cell lung cancer A549 and H1299 cells
title: 升陷汤含药血清抑制非小细胞肺癌A549、H1299细胞恶性生物学行为研究

description: Objective To investigate the effects of Shengxian Tang on the biological behavior of non-small cell lung cancer cells, specifically A549 and H1299.Methods A549 and H1299 non-small cell lung cancer cells in the logarithmic growth phase were cultured in serum containing Shengxian Tang at low (3 mg/mL), medium (6 mg/mL), and high (9 mg/mL) concentrations. The activity and proliferation capacity of A549 and H1299 cells were evaluated using the CCK-8 method. The clonal formation ability of A549 and H1299 cells was assessed using a colony formation assay. The migration ability of A549 and H1299 cells was evaluated using a scratch assay. The migration and invasion ability of A549 and H1299 cells were detected by Transwell. The cell cycles of A549 and H1299 were detected by flow cytometry. The expression levels of various proteins were measured using Western Blot analysis, including apoptosis-related proteins (Bcl-2, Caspase-3, Caspase-9), autophagy-related proteins (LC3B, p62), epithelial-mesenchymal transition-related proteins (E-cadherin, FSP-1, α-SMA), and angiogenesis-related proteins (CD34, VEGFA). The fluorescence intensity of A549, H1299 angiogenic proteins (CD31, CD34) was evaluated by immunofluorescence.Results Compared with the control group, the low, medium, and high doses of Shengxian Tang-containing serum significantly inhibited the proliferation, clonal formation, migration, and invasion of A549 and H1299 cells. Additionally, high doses of Shengxian Tang-containing serum significantly increased the proportion of cells in the G2/M phase of the cell cycle. After the intervention of low, medium and high doses of Shengxian Tang-containing serum, the number of apoptosis of A549 cells was significantly increased, and the protein expression levels of Bcl-2, Caspase-3 and Caspase-9 were significantly decreased. In addition, the expression level of P62 in A549 cells was significantly increased and the ratio of LC3BⅡ/I was significantly decreased in low and medium doses of Shengxian Tang-containing serum, while the expression level of p62 and LC3BⅡ/I was significantly decreased in high doses of Shengxian Tang-containing serum. In H1299 cells, the expression level of p62 was significantly decreased by the high-dose serum containing Shengxian Tang. Low, medium, and high doses of Shengxian Tang-containing serum significantly increased the expression levels of E-cadherin and FSP-1 in A549 cells. Medium and high doses significantly decreased the protein expression level of α-SMA. For H1299 cells, the expression of E-cadherin protein was increased by medium and high doses of Shengxian Tang, while α-SMA protein expression was decreased by high doses. The expression levels of VEGFA and CD34 in A549 cells were significantly decreased by low, medium and high doses of Shengxian Tang. For H1299 cells, low and medium doses decreased VEGFA protein expression, while high doses decreased CD34 protein expression. Immunofluorescence experiments confirmed that treatment with Shengxian Tang significantly decreased the fluorescence intensity of CD31 and CD34 in both A549 and H1299 cells.Conclusion Shengxian Tang significantly inhibits the proliferation, colony formation, migration, invasion, and epithelial-mesenchymal transition (EMT) of NSCLC A549 and H1299 cells, while also delaying the cell cycle. Additionally, it reduces autophagy levels in A549 cells and promotes their apoptosis.
description: Objective To investigate the effects of Shengxian Tang on the biological behavior of non-small cell lung cancer cells, specifically A549 and H1299.Methods A549 and H1299 non-small cell lung cancer cells in the logarithmic growth phase were cultured in serum containing Shengxian Tang at low (3 mg/mL), medium (6 mg/mL), and high (9 mg/mL) concentrations. The activity and proliferation capacity of A549 and H1299 cells were evaluated using the CCK-8 method. The clonal formation ability of A549 and H1299 cells was assessed using a colony formation assay. The migration ability of A549 and H1299 cells was evaluated using a scratch assay. The migration and invasion ability of A549 and H1299 cells were detected by Transwell. The cell cycles of A549 and H1299 were detected by flow cytometry. The expression levels of various proteins were measured using Western Blot analysis, including apoptosis-related proteins (Bcl-2, Caspase-3, Caspase-9), autophagy-related proteins (LC3B, p62), epithelial-mesenchymal transition-related proteins (E-cadherin, FSP-1, α-SMA), and angiogenesis-related proteins (CD34, VEGFA). The fluorescence intensity of A549, H1299 angiogenic proteins (CD31, CD34) was evaluated by immunofluorescence.Results Compared with the control group, the low, medium, and high doses of Shengxian Tang-containing serum significantly inhibited the proliferation, clonal formation, migration, and invasion of A549 and H1299 cells. Additionally, high doses of Shengxian Tang-containing serum significantly increased the proportion of cells in the G2/M phase of the cell cycle. After the intervention of low, medium and high doses of Shengxian Tang-containing serum, the number of apoptosis of A549 cells was significantly increased, and the protein expression levels of Bcl-2, Caspase-3 and Caspase-9 were significantly decreased. In addition, the expression level of P62 in A549 cells was significantly increased and the ratio of LC3BⅡ/I was significantly decreased in low and medium doses of Shengxian Tang-containing serum, while the expression level of p62 and LC3BⅡ/I was significantly decreased in high doses of Shengxian Tang-containing serum. In H1299 cells, the expression level of p62 was significantly decreased by the high-dose serum containing Shengxian Tang. Low, medium, and high doses of Shengxian Tang-containing serum significantly increased the expression levels of E-cadherin and FSP-1 in A549 cells. Medium and high doses significantly decreased the protein expression level of α-SMA. For H1299 cells, the expression of E-cadherin protein was increased by medium and high doses of Shengxian Tang, while α-SMA protein expression was decreased by high doses. The expression levels of VEGFA and CD34 in A549 cells were significantly decreased by low, medium and high doses of Shengxian Tang. For H1299 cells, low and medium doses decreased VEGFA protein expression, while high doses decreased CD34 protein expression. Immunofluorescence experiments confirmed that treatment with Shengxian Tang significantly decreased the fluorescence intensity of CD31 and CD34 in both A549 and H1299 cells.Conclusion Shengxian Tang significantly inhibits the proliferation, colony formation, migration, invasion, and epithelial-mesenchymal transition (EMT) of NSCLC A549 and H1299 cells, while also delaying the cell cycle. Additionally, it reduces autophagy levels in A549 cells and promotes their apoptosis.