Data from: Loss of miR-143 and miR-145 in condyloma acuminatum promotes cellular proliferation and inhibits apoptosis by targeting NRAS

The expression profile of miRNAs and their function in condyloma acuminatum (CA) remains unknown. In this study, we aimed to detect the effects of miR-143 and miR-145, the most downregulated in CA samples using high-throughput sequencing, on cell proliferation and apoptosis, to determine a novel therapeutic target for CA recurrence. RT-qPCR was used to validate the lower expression of miR-143 and miR-145 in a larger size of CA samples, and the expression of NRAS in CA samples was significantly higher than self-controls as determined western blotting assay. Luciferase assay was performed to confirm that miR-143 or miR-145 targeted NRAS directly. Transduction of LV-pre-miR-143 or LV-pre-miR-145 to human papilloma virus (HPV)-infected SiHa cells led to reduced proliferation, greater apoptosis, and inhibition of expression of NRAS, PI3Kp110α, and pAKT. However, knockout of miR-143 or miR-145 in human epidermal keratinocytes (HEKs) by delivery of CRISPR/CAS9-gRNA for target miRNAs protected cells from apoptosis and upregulated expression of target genes as described above. MiR-143 and miR-145 sensitized cells to nutlin-3a, a p53 activator and MDM2 antagonist, while their loss protected cells from the stress of nutlin-3a. Furthermore, siRNA targeting NRAS showed similar effects on proliferation and apoptosis as miR-143 or miR-145. Taken together, our results suggest that loss of miR-143 or miR-145 in CA protects HPV-infected cells from apoptosis induced by environmental stress, in addition to promoting cellular proliferation and inhibiting apoptosis by targeting NRAS/PI3K/ATK. Restoration of miR-143 or miR-145 might provide an applicable and novel approach to block the recurrence and progression of CA.

尖锐湿疣（condyloma acuminatum, CA）组织中微小RNA（microRNA, miRNA）的表达谱及其功能尚未明确。本研究旨在探究经高通量测序筛选得到的CA样本中下调最为显著的miR-143与miR-145对细胞增殖与凋亡的调控作用，以期为CA复发探寻全新的治疗靶点。我们采用实时定量聚合酶链式反应（RT-qPCR）在更大样本量的CA组织中验证了miR-143与miR-145的低表达水平；同时通过蛋白质印迹实验发现，CA样本中NRAS的表达量显著高于自身对照样本。双荧光素酶报告基因实验证实，miR-143或miR-145可直接靶向NRAS。将携带pre-miR-143或pre-miR-145的慢病毒（LV）转染人乳头瘤病毒（HPV）感染的SiHa细胞后，细胞增殖能力降低、凋亡水平显著升高，且NRAS、PI3Kp110α及pAKT的表达受到明显抑制。而通过CRISPR/CAS9-gRNA系统靶向敲除人表皮角质形成细胞（HEKs）中的miR-143或miR-145，则可使细胞抵抗凋亡，并上调上述靶基因的表达。miR-143与miR-145可增强细胞对nutlin-3a（一种p53激活剂与MDM2拮抗剂）的敏感性，而二者的敲除则可使细胞免受nutlin-3a诱导的应激损伤。此外，靶向NRAS的小干扰RNA（siRNA）可产生与miR-143或miR-145类似的增殖与凋亡调控效应。综上，本研究结果表明，CA组织中miR-143或miR-145的缺失，不仅可通过靶向NRAS/PI3K/AKT通路促进细胞增殖、抑制凋亡，还可使HPV感染的细胞免受环境应激诱导的凋亡。恢复miR-143或miR-145的表达，或许可为阻断CA的复发与进展提供一种可行的全新治疗策略。