Knockdown of the zinc transporter protein ZIP7 to inhibit the proliferation of mouse testicular sertoli cells
Objective:To investigate the effects and its possible mechanism of knocking down the zinc transporter protein ZIP7 on the proliferation of mouse testicular sertoli cells.Methods:Experiments were carried out using a mouse testicular sertoli cell line(TM4 cells).The subcellular localization of ZIP7 in TM4 cells was observed by cellular immunofluorescence staining after the culture of TM4 cells.TM4 cells were divided into the control group and the ZIP7 siRNA group(siRNA transfection was used to knock down the expression of ZIP7 in TM4 cells).The proliferation viability of the two groups were detected by CCK8 assay,and the levels of reactive oxygen species(ROS)in the two groups were detected using a DHE fluorescent probe.The protein expression levels of ZIP7,c-Jun amino-terminal kinase(JNK),phosphorylated J NK(p-JNK),extracellular signal-regulated kinase(ERK)and phosphorylated ERK(p-ERK)in the cells of both groups were detected by Western blot.Results:The results of cellular immunofluorescence staining showed that ZIP7 was located in the endoplasmic reticulum of TM4 cells.The siRNA interference resulted in significantly lower mRNA and protein expressions of ZIP7 in ZIP7 siRNA group than those in the control group(P＜0.05).After transfection with siRNA and knockdown of ZIP7 expression in TM4 cells,the proliferative viability of cells in the ZIP7 siRNA group was significantly lower than that in the control group,while the intracellular ROS level was significantly higher than that of the control group(P＜0.001).Compared with the control group,there was no significant difference in the expression of p-ERK/ERK protein in the cells of the ZIP7 siRNA group(P＞0.05),while the expression of p-JNK/JNK protein was significantly increased(P＜0.05).Conclusions:Knockdown of ZIP7 can significantly inhibited the proliferation of TM4 cells.The process may be mediated through elevated ROS levels and activation of JNK phosphorylation.
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