Objective:To investigate the chemical components of Semen podocarpi extract(SPE)and its effect on nasopharyngeal carcinoma cells and CNE-2R cells.Methods:Chemical components in SPE were identified by UPLC-MS/MS.CCK-8 ...Objective:To investigate the chemical components of Semen podocarpi extract(SPE)and its effect on nasopharyngeal carcinoma cells and CNE-2R cells.Methods:Chemical components in SPE were identified by UPLC-MS/MS.CCK-8 and cell cloning experiments were applied to evaluate the effects of SPE on the proliferation of CNE-2R cells,and a single-hit multitarget model was used to calculate the radiobiological parameters.Cell apoptosis and cell cycle were analyzed by flow cytometry,and the levels of genes and proteins of the Raf/MEK/ERK pathway were determined by RT-PCR and Western blotting.Results:A total of 37 compounds from SPE were identified,and SPE with or without irradiation inhibited the proliferation of CNE-2R cells.SPE also promoted apoptosis,arrested cells in the G_(2)/M phase,and presented radiosensitizing effects.Compared with irradiation alone,the effects of SPE+irradiation on apoptosis and cell cycle distribution were not significantly different.In addition,SPE had no significant effect on MEK gene expression.SPE significantly increased the gene expression of C-Raf and significantly reduced the protein expression of C-Raf,as well as the gene and protein expression of ERK1 and ERK2.The protein levels of C-Raf,ERK1,and ERK2 were also significantly lower in cells treated with SPE+irradiation than in cells treated with irradiation alone.Conclusions:The effects of SPE on inhibiting cell proliferation and promoting apoptosis are likely associated with cell cycle arrest and Raf/MEK/ERK pathway regulation,and the mechanism underlying radiosensitization by SPE may involve downregulating the protein expression of C-Raf,ERK1,and ERK2.展开更多
基金supported by the Project of Administration of Traditional Chinese Medicine of Guangxi Zhuang Autonomous Region(grant number GZSY22-69)the Middle/Young aged Teachers’Research Ability Improvement Project of Guangxi Higher Education(grant number 2024KY0120).
文摘Objective:To investigate the chemical components of Semen podocarpi extract(SPE)and its effect on nasopharyngeal carcinoma cells and CNE-2R cells.Methods:Chemical components in SPE were identified by UPLC-MS/MS.CCK-8 and cell cloning experiments were applied to evaluate the effects of SPE on the proliferation of CNE-2R cells,and a single-hit multitarget model was used to calculate the radiobiological parameters.Cell apoptosis and cell cycle were analyzed by flow cytometry,and the levels of genes and proteins of the Raf/MEK/ERK pathway were determined by RT-PCR and Western blotting.Results:A total of 37 compounds from SPE were identified,and SPE with or without irradiation inhibited the proliferation of CNE-2R cells.SPE also promoted apoptosis,arrested cells in the G_(2)/M phase,and presented radiosensitizing effects.Compared with irradiation alone,the effects of SPE+irradiation on apoptosis and cell cycle distribution were not significantly different.In addition,SPE had no significant effect on MEK gene expression.SPE significantly increased the gene expression of C-Raf and significantly reduced the protein expression of C-Raf,as well as the gene and protein expression of ERK1 and ERK2.The protein levels of C-Raf,ERK1,and ERK2 were also significantly lower in cells treated with SPE+irradiation than in cells treated with irradiation alone.Conclusions:The effects of SPE on inhibiting cell proliferation and promoting apoptosis are likely associated with cell cycle arrest and Raf/MEK/ERK pathway regulation,and the mechanism underlying radiosensitization by SPE may involve downregulating the protein expression of C-Raf,ERK1,and ERK2.
