Huaier Aqueous Extract Mediates Keloid Cell Proliferation and Apoptosis by Regulating miR-128-3p / EPHB2
Huaier Aqueous Extract Mediates Keloid Cell Proliferation and Apoptosis by Regulating miR-128-3p / EPHB2
Xiaoguang Su1,*, Yanjun Gao2, Yanling Wang1 and Yaohui Ma2
ABSTRACT
This study was aimed to investigate the effect of HAE on the proliferation and apoptosis of keloid fibroblasts and its mechanism. Primary HFB cells were isolated and cultured, and treated with different concentrations of HAE for 48 h. Cell viability was measured by MTT. Flow cytometry was used to detect the apoptosis rate. qRT-PCR and Western blot were used to detect the effects of HAE on miR-128-3p and EPHB2. The above method was used to detect cell viability and apoptosis rate after overexpression of miR-128-3p or inhibition of EPHB2. The dual luciferase report experiment verified the targeted binding relationship between miR-128-3p and EPHB2. Western blot was used to detect the expression of Ki-67, PCNA, Bcl-2 and Bax. Our findings showed that after HAE treatment, the cell viability was significantly reduced (P <0.05), the apoptosis rate was significantly increased (P <0.05), the levels of Ki-67, PCNA, Bcl-2 protein were significantly reduced (P <0.05), and the level of Bax protein was significantly increased (P <0.05), the expression level of miR-128-3p was increased significantly (P <0.05), the expression levels of EPHB2 mRNA and protein levels were decreased significantly (P <0.05). After overexpression of miR-128-3p or inhibition of EPHB2, the cell viability was significantly reduced (P <0.05), the apoptosis rate was significantly increased (P <0.05), and the levels of Ki-67, PCNA, and Bcl-2 protein were significantly reduced (P <0.05), the level of Bax protein was increased significantly (P <0.05). Double luciferase reporting experiments confirmed that miR-128-3p could target EPHB2 and negatively regulate its expression and activity. We concluded HAE may inhibit keloid fibroblast proliferation and promote apoptosis by regulating the miR-128-3p / EPHB2 molecular axis.
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