piRNA-14633 promotes cervical cancer cell malignancy in a METTL14-dependent m6A RNA methylation manner

Abstract

Background Cervical cancer (CC) is one of the most common gynecological tumors that threatens women's health and lives. Aberrant expression of PIWI-interacting RNA (piRNA) is closely related with a range of cancers and can serve as a tumor promoter or suppressor in proliferation, migration and invasion. In this study, the aim was not only to discover differential expression of piRNA in CC tissue (CC cells) and normal cervical tissue (normal cervical epithelium cells), but also to investigate the biological function and action mechanism of piRNA in CC. Methods The DESeq2 approach was used to estimate fold change in piRNA between CC tissue and normal cervical tissue. The relative expressions of piRNAs (piRNA-20657, piRNA-20497, piRNA-14633 and piRNA-13350) and RNA m6A methyltransferases/demethylases were detected using RT-qPCR. After intervention with piRNA-14633 and METTL14 expression, the viability of CaSki cells and SiHa cells was detected by CCK8. CC cell proliferation was detected by colony formation assay. Apoptosis rate and cell cycle were detected by flow cytometry. Transwell assay was performed to detect cell migration and invasion. EpiQuik m6A RNA Methylation Quantification Kit was used to evaluate m6A RNA methylation levels. Expression of methyltransferase-like protein 14 (METTL14), PIWIL-proteins and CYP1B1 were detected by RT-qPCR and western blot. The effect of piRNA-14633 on METTL14 was evaluated by a dual-luciferase reporter assay. The in vivo effects of piRNA-14633 on CC was assessed by nude mice experiments. Results piRNA-14633 showed high expression in CC tissues and cells, piRNA-14633 mimic (piRNA-14633 overexpression) promoted viability, proliferation, migration and invasion of CaSki cells and SiHa cells. Besides, piRNA-14633 mimic increased m6A RNA methylation levels and METTL14 mRNA stability. Results of dual luciferase reporter assays indicated that METTL14 was a directed target gene of piRNA-14633. Knockdown of METTL14 with siRNA attenuated proliferation, migration and invasion of CC cells. piRNA-14633 increased CYP1B1 expression, while silencing of METTL14 impaired its expression. The effect of piRNA overexpression on METTL14 expression has concentration-dependent characteristics. Results from in vivo experiment indicated that piRNA-14633 promoted cervical tumor growth. Conclusion piRNA-14633 promotes proliferation, migration and invasion of CC cells by METTL14/CYP1B1 signaling axis, highlighting the important role of piRNA-14633 in CC.

YTHDF3 mediates HNF1α regulation of cervical cancer radio‐resistance by promoting RAD51D translation in an m6A‐dependent manner

“…For example, m6A METTL3 promotes cervical cancer tumourigenesis and the Warburg effect through YTHDF1/HK2 modification [34]. piRNA‐14633 promotes malignancy in cervical cancer cells in a METTL14‐dependent m6A RNA methylation [35]. FTO reduces β‐catenin expression by reducing m6A in its mRNA transcripts, thereby enhancing chemoradiotherapy resistance [36].…”

The epigenetic regulatory mechanism of PIWI/piRNAs in human cancers

“…And here are some related researches as followed. Xie’s research shows that piRNA-14633 can facilitate the malignancy in cervical cancer (CC) by piRNA-14633/METTL14/CYP1B1 axis [ 84 ]. In this study, piRNA-14633 is highly expressed in CC cells, and it raises m 6 A RNA methylation levels and METTL14 mRNA stability.…”

RNA Modifications Meet Tumors

“…For example, regarding m 6 A modification, METTL14 plays the role of tumor inhibitor in CRC, 168 , 169 bladder cancer, 170 and breast cancer. 171 It is a carcinogenic factor in cervical cancer, 172 EOC, 173 pancreatic cancer 174 and leukemia 175 , 176 during their development and incidence. 177 , 178 Therefore, studies on RNA modifications should be carried out extensively and comprehensively in various types of tumors.…”