ARFHPV E7 oncogene, lncRNA HOTAIR, miR‐331‐3p and its target, NRP2, form a negative feedback loop to regulate the apoptosis in the tumorigenesis in HPV positive cervical cancer

摘要

Abstract The objective of this study was to explore the role of HOTAIR in the development of cervical cancer, as well as its downstream signaling pathway. We conducted computational analysis, luciferase assay to explore downstream of HOTAIR and miR‐331‐3p. Real‐time PCR and Western blot were carried out to detect the relationship among E7, HOTAIR, miR‐331‐3p, NRP2, and P53. Finally, MTT assay and flow cytometry analysis were performed to validate the effect of E7 and miR‐331‐3p on cell apoptosis and proliferation. NRP2 was identified as a virtual target gene of miR‐331‐3p with a binding site of miR‐331‐3p, and HOTAIR was directly sponged to miR‐331‐3p, miR‐331‐3p reduced luciferase activity of wild‐type of NRP2 3′UTR and HOTAIR, but not those of mutant NRP2 3′UTR and HOTAIR. MiR‐331‐3p down‐regulated NRP2 and E7 expression levels, and further promoted cell apoptosis, while inhibited cell proliferation. Cell transfected with HPV16 E7 displayed lower levels of HOTAIR, NRP2 and P53, a higher level of miR‐331‐3p, over‐expression of E7 further repressed cell apoptosis, while improved cell proliferation compared with control. Normal HPV (+) group exhibited a higher miR‐331‐3p, and lower mRNA levels of HOTAIR and NRP2 than HPV (?) group. According to the result of IHC (immunohistochemistry), we found that NRP2 protein was highly expressed in HPV (?) group compared to that in HPV (+) group. E7‐HOTAIR‐miR‐331‐3p‐NRP2‐E7 formed a regulatory loop, and could be involved in the pathogenesis of cervical cancer.