Euphorbia helioscopia inhibits proliferation, invasion, and migration and promotes apoptosis of non-small cell lung cancer cells

doi:10.12122/j.issn.1673-4254.2024.10.10

Abstract

Abstract:

Objective To investigate the effect of Euphorbia helioscopia on biological behaviors of non-small cell lung cancer (NSCLC) cells. Methods NSCLC cell lines PC-9 and A549 treated with different concentrations of Euphorbia helioscopia preparations were examined for changes in proliferation, apoptosis, invasion and migration using CCK-8 assay, colony formation assay, flow cytometry, wound healing assay and Transwell assay. Western blotting was performed to detect the changes in protein expressions of Bax, Bcl-2, E-cadherin, vimentin, MMP2, and MMP9 in the treated cells. PC-9 cells were injected subcutaneously into BALB/C nude mice to establish a nude mouse subcutaneous tumor model. According to the growth of subcutaneous tumors, mice were randomly divided into control group: gavaged daily with saline; Euphorbia helioscopia-treated group: gavaged daily with Euphorbia helioscopia 65 mg/mL, and Euphorbia helioscopia granules were dissolved in saline; cisplatin-treated group: injected intraperitoneally with cisplatin 4 mg/kg every 5 days, 6 mice per group. The subcutaneous tumor volume and mass changes of mice were measured, and the toxic effects of Euphorbia helioscopia on heart, liver, spleen, lung and kidney as well as the therapeutic effects of Euphorbia helioscopia were observed in the mice bearing tumor. Results Euphorbia helioscopia granules concentration-dependently inhibited the proliferation and survival of PC-9 and A549 cells, significantly promoted cell apoptosis, suppressed invasion and migration abilities of the cells, up-regulated the expression levels of E-cadherin and Bax, and down-regulated the expressions of Bcl-2, vimentin, MMP2, and MMP9. In the tumor-bearing mice, treatment with Euphorbia helioscopia significantly inhibited tumor growth without producing obvious toxicity in the vital organs. Conclusion Euphorbia helioscopia can inhibit proliferation, invasion, and migration and induces apoptosis of NSCLC cells in vitro.

Key words: Euphorbia helioscopia, non-small cell lung cancer, proliferation, apoptosis, invasion and migration

Xuerou LIU, Yumei YANG, Wei LIU, Zhen ZHANG, Xingqi ZHOU, Wenyu XIE, Lin SHEN, Mengxiao ZHANG, Xian LI, Jialan ZANG, Shanshan LI. Euphorbia helioscopia inhibits proliferation, invasion, and migration and promotes apoptosis of non-small cell lung cancer cells[J]. Journal of Southern Medical University, 2024, 44(10): 1918-1925.

Figures/Tables 5

Fig.1 Euphorbia helioscopia inhibits proliferation of PC-9 and A549 cells. A: CCK-8 assay. B: Colony formation assay of PC-9 and A549 cells. C: Morphological changes of PC-9 and A549 cells (scale bar=200 μm). **P<0.01 vs 0 mg/mL.

Fig.2 Euphorbia helioscopia induces apoptosis in PC-9 and A549 cells. A: Apoptosis of PC-9 cells treated with different concentrations of Euphorbia helioscopia (0, 0.4, 0.8, and 1.2 mg/mL) detected by flow cytometry. B: Apoptosis of A549 cells treated with different concentrations of Euphorbia helioscopia (0, 0.8, 1.2, 1.6 mg/mL) detected by flow cytometry. C: Expressions of Bcl-2 and Bax in PC-9 cells detected by Western blotting. D: Expressions of Bcl-2 and Bax in A549 cells detected by Western blotting. *P<0.05, **P<0.01 vs 0 mg/mL.

Fig.3 Euphorbia helioscopia inhibits invasion and migration of PC-9 cells. A: Wound healing assay of PC-9 cells treated with different concentrations of Euphorbia helioscopia. B: Crystal violet staining of the cells (scale bar=200 μm). C: Numbers of PC-9 cells passing through Transwell chamber. D, E: Expressions of E-cadherin, vimentin, MMP2, and MMP9 in PC-9 cells detected by Western blotting. *P<0.05, **P<0.01 vs 0 mg/mL.

Fig.4 Euphorbia helioscopia inhibits invasion and migration of A549 cells. A: Wound healing assay of A549 cells treated with different concentrations of Euphorbia helioscopia. B: Crystal violet staining of the cells (scale bar=200 μm). C: Numbers of A549 cells passing through Transwell chamber. D, E: Expressions of E-cadherin, vimentin, MMP2, and MMP9 in A549 cells detected by Western blotting. *P<0.05, **P<0.01 vs 0 mg/mL.

Fig.5 Euphorbia helioscopia inhibits growth of PC-9 cell xenografts in nude mice. A: Tumor growth in each group. B: Tumor weight in the 3 groups. C: Tumor size in the 3 groups. D: Body weight changes of the mice. E: Heart, liver, spleen, lung, and kidney tissues of the mice in the 3 groups. **P<0.01 vs control.

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