Gang Yu1, Lizhen Chen2, Yuanhua Hu3, Zhen Yuan4, Yao Luo5, Yuanhuan Xiong1,6. 1. Jiangxi Provincial People's Hospital, Nanchang, China. 2. Jingdezhen No. 1 People's Hospital, Jingdezhen, China. 3. Shunde Maternal and Child Health Hospital, Guangdong Medical University, Foshan, China. 4. Yichun Institute of Vocational Technology, Yichun, China. 5. Medical College of Nanchang University, Nanchang, China. 6. Jiangxi the Fifth People's Hospital, Nanchang, China.
Abstract
BACKGROUND: Due to dual-regulating carcinogenesis, the TGFβ pathway is an ideal and alternative tumor target. Natural flavonoids possess the similar structures to estrogen and could exert an important benefit to cervical cancer. The present study aimed to screen the inhibitor of TGFβ pathway from natural flavonoids and evaluate the function and mechanism of the TGFβ pathway inhibitor on cervical cancer. MATERIALS AND METHODS: The cervical cancer HeLa cells were firstly treated with different flavonoids and probed by western blot for screening the inhibitor of TGFβ pathway. And then, the effect of the identified inhibitor on cell proliferation was studied by CCK-8 and clone formation assay. Then, RT-PCR and western blot assay were performed to evaluate the effect of identified inhibitor on mTOR/p70S6K pathway, and the cell migration and EMT pathway were also examined using scratching analysis and western blot assay. Finally, the role of TGFβ was assessed via the classic inhibitor of TGFβ/SMAD pathway. RESULTS: Screening data by western blot assay showed that baicalein displayed the best inhibitor effect on TGFβ expression. CCK-8 and clone formation assay showed that baicalein inhibited the cell proliferation and clone cell number. RT-PCR and western bolt for probing mTOR, p70S6K, and 4EBP1 revealed that baicalein could suppress their expression and phosphorylation. The scratching analysis and western blot assay displayed that baicalein inhibited the cell migration and EMT progression in HeLa. The use of SB431542, a TGFβ inhibitor, revealed that TGFβ was crucial to baicalein-regulating cell proliferation and migration in HeLa cells. CONCLUSION: Baicalein, a medicine agent screened from natural flavonoids targeting TGFβ pathway, could suppress mTOR/p70S6K pathway-mediated cell proliferation and EMT pathway-related migration via TGFβ pathway in cervical cancer HeLa cells.
BACKGROUND: Due to dual-regulating carcinogenesis, the TGFβ pathway is an ideal and alternative tumor target. Natural flavonoids possess the similar structures to estrogen and could exert an important benefit to cervical cancer. The present study aimed to screen the inhibitor of TGFβ pathway from natural flavonoids and evaluate the function and mechanism of the TGFβ pathway inhibitor on cervical cancer. MATERIALS AND METHODS: The cervical cancer HeLa cells were firstly treated with different flavonoids and probed by western blot for screening the inhibitor of TGFβ pathway. And then, the effect of the identified inhibitor on cell proliferation was studied by CCK-8 and clone formation assay. Then, RT-PCR and western blot assay were performed to evaluate the effect of identified inhibitor on mTOR/p70S6K pathway, and the cell migration and EMT pathway were also examined using scratching analysis and western blot assay. Finally, the role of TGFβ was assessed via the classic inhibitor of TGFβ/SMAD pathway. RESULTS: Screening data by western blot assay showed that baicalein displayed the best inhibitor effect on TGFβ expression. CCK-8 and clone formation assay showed that baicalein inhibited the cell proliferation and clone cell number. RT-PCR and western bolt for probing mTOR, p70S6K, and 4EBP1 revealed that baicalein could suppress their expression and phosphorylation. The scratching analysis and western blot assay displayed that baicalein inhibited the cell migration and EMT progression in HeLa. The use of SB431542, a TGFβ inhibitor, revealed that TGFβ was crucial to baicalein-regulating cell proliferation and migration in HeLa cells. CONCLUSION: Baicalein, a medicine agent screened from natural flavonoids targeting TGFβ pathway, could suppress mTOR/p70S6K pathway-mediated cell proliferation and EMT pathway-related migration via TGFβ pathway in cervical cancer HeLa cells.