17 January 2020 : Laboratory Research
Downregulation of SEMA4C Inhibit Epithelial-Mesenchymal Transition (EMT) and the Invasion and Metastasis of Cervical Cancer Cells via Inhibiting Transforming Growth Factor-beta 1 (TGF-β1)-Induced Hela cells p38 Mitogen-Activated Protein Kinase (MAPK) ActivationLilan Yang12ABCDEFG, Yayuan Yu1ABCF, Zhenfang Xiong3BC, Hongxia Chen2BC, Buzhen Tan1ADG*, Hui Hu1ADG
Med Sci Monit 2020; 26:e918123
BACKGROUND: Epithelial-mesenchymal transition (EMT) plays a key role in promoting invasion and metastasis of tumor cells. SEMA4C can regulate the generation of transforming growth factor-beta 1 (TGF-β1)-induced EMT in cervical cancer. This study investigated the relationship between the regulation of SEMA4C on TGF-β1-induced p38 mitogen-activated protein kinase (MAPK) activation and invasion and metastasis of cervical cancer.
MATERIAL AND METHODS: Hela-shSEMA4C cell line was established and the success of transfection was confirmed with fluorescence intensity. Cell experiments were divided into 2 groups. Group 1 was Hela, Hela-shNC, and Hela-shSEMA4C; and Group 2 was Hela, Hela-shNC, Hela-shSEMA4C, Hela+TGF-β1, Hela-shNC+TGF-β1, and Hela-shSEMA4C+TGF-β1. Group 1 was detected for SEMA4C mRNA expression by real-time polymerase chain reaction (RT-PCR), cell viability by Cell Counting Kit-8 (CCK-8), F-actin fluorescence intensity by immunofluorescence, cell migration by scratch test, and cell invasion by invasion test. Group 2 was analyzed for E-cadherin fluorescence intensity by immunofluorescence, human fibronectin (FN) content by enzyme-linked immunosorbent assay (ELISA), and SEMA4C, E-cadherin and p-p38 expressions by Western blot.
RESULTS: For Group 1, compared with Hela and Hela-shNC subgroups, the SEMA4C mRNA expression, cell viability, F-actin fluorescence intensity, cell migration and invasion ability in the Hela-shSEMA4C subgroup were significantly decreased (P<0.05). For Group 2, compared with Hela and Hela-shNC subgroups, the E-cadherin expression and fluorescence intensity in the Hela-shSEMA4C subgroup were significantly increased (P<0.01), while the FN content, SEMA4C, and p-p38 MAPK expressions were significantly decreased (P<0.01). Compared with Hela-shNC+TGF-β1 and Hela+TGF-β1 subgroups, the E-cadherin expression and fluorescence intensity in the Hela-shSEMA4C+TGF-β1 subgroup were significantly increased (P<0.01), while the FN content, SEMA4C and p-p38 expressions were significantly decreased (P<0.01).
CONCLUSIONS: Downregulation of SEMA4C can inhibit EMT and the invasion and metastasis of cervical cancer cells via inhibiting TGF-β1-induced Hela cells p38 MAPK activation.
Keywords: Neoplasm Invasiveness, Neoplasm Metastasis, Semaphorins, Transforming Growth Factor beta, Uterine Cervical Neoplasms, Cadherins, Cell Movement, Cell Survival, Down-Regulation, Enzyme Activation, Epithelial-mesenchymal transition, Fibronectins, Fluorescence, Hela Cells, Phosphorylation, RNA, Messenger, RNA, Small Interfering, Transforming Growth Factor beta1
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