Effects Of EN2(Engrailed-2) RNA Interference On Biological Behavior Of Human Renal Tubular Epithelial Cell

Obiective: Our preliminary study indicated that Engrailed-2(EN2) is downregulated but also ectopically expressed in clear-cell renal cell carcinoma(CCRCC), and the absence of EN2 expression was associated with poor histological grade. However, the specific roles of EN2 in CCRCC have yet to be elucidated. In the present study, we examined the effects of inhibiting En-2 expression in HK-2 to figure out how does EN2 gene regulate cell functions of HK-2.Methods: Lentivirus small hairpin RNA(shRNA) interference techniques were utilized to effect EN2 gene’s expression of HK-2 cell line. Then quantitative real-time polymerase chain reaction(RT-qPCR) and western blotting were applied to quantify EN2 mRNA and protein expression level. MTT assay was conducted to measure cell proliferation activity.Cell cycle and apoptosis were detected by flow cytometry, invasive efficiency was accessed by transwell invasion test and wound scratch assay applied for testing cell migration ability.Results: Lentivirus-mediated shRNA significantly inhibitied the expression of EN2 in HK-2 cell line at both mRNA and protein level. Compared with Lv-shcon and control group, cell proliferation of Lv-shEN2 speeded up. The treatment shortened the cell cycle,lessened cell apoptosis and made the Lv-shEN2 acted more invasively. However, there was no difference among the migration ability of three groups.Conclusion: EN2 might play an anti-oncogenic role in oncogenesis and maintaining the normal growth of human renal tubular epithelial cells.