MicroRNA-18a targeting of the STK4/MST1 tumour suppressor is necessary for transformation in HPV positive cervical cancer

Human papillomaviruses (HPV) are a major cause of malignancy worldwide. They are the aetiological agents of almost all cervical cancers as well as a sub-set of other anogenital and head and neck cancers. Hijacking of host cellular pathways is essential for virus pathogenesis; however, a major challenge remains to identify key host targets and to define their contribution to HPV-driven malignancy. The Hippo pathway regulates epithelial homeostasis by down-regulating the function of the transcription factor YAP. Increased YAP expression has been observed in cervical cancer but the mechanisms driving this increase remain unclear. We found significant down-regulation of the master Hippo regulatory kinase STK4 (also termed MST1) in cervical disease samples and cervical cancer cell lines compared with healthy controls. Re-introduction of STK4 inhibited the proliferation of HPV positive cervical cells and this corresponded with decreased YAP nuclear localization and decreased YAP-dependent gene expression. The HPV E6 and E7 oncoproteins maintained low STK4 expression in cervical cancer cells by upregulating the oncomiR miR-18a, which directly targeted theSTK4mRNA 3'UTR. Interestingly, miR-18a knockdown increased STK4 expression and activated the Hippo pathway, significantly reducing cervical cancer cell proliferation. Our results identify STK4 as a key cervical cancer tumour suppressor, which is targeted via miR-18a in HPV positive tumours. Our study indicates that activation of the Hippo pathway may offer a therapeutically beneficial option for cervical cancer treatment. Author summary HPVs are the causative agents of similar to 5% of human cancers. Better understanding of the mechanisms by which these viruses deregulate cellular signalling pathways may offer therapeutic options for HPV-associated malignancies. The transcription factor YAP is active in cervical cancer but the mechanisms controlling its activation remain unclear. YAP is negatively regulated and sequestered in the cytoplasm through activation of the Hippo pathway. We discovered that expression of the master Hippo kinase, STK4 (also termed MST1), is reduced in HPV positive cervical cell lines and cervical disease samples. Low STK4 levels were maintained by the HPV oncogenes through up-regulation of miR-18a, which targeted theSTK4mRNA 3'UTR. Re-introduction of STK4 or bypassing miR-18a-dependent regulation de-activated YAP-driven transcription and reduced cell proliferation. Thus, our study identifies a novel interplay between HPV oncogenes and the STK4 tumour suppressor and identifies the Hippo pathway as a target for therapeutic intervention in HPV-associated malignancies.