Expression level of miR-196a in patients with HPV16 and HPV18 subtypes infections and bioinformatics analysis of its association with survival of cervical cancer

OBJECTIVE To explore the expression level of miR-196a in cervical cells infected with high-risk human papillomavirus (HPV) 16 and 18.

METHODS The Gene Expression Omnibus (GEO) was used to screen for differentially expressed miRNAs between HPV 16 or 18-positive cervical cancer cells and normal cervical cells. Online biological software https://kmplot.com/analysis/ was utilized to analyze the relationship between the most differentially expressed miRNA and the overall survival of cervical cancer patients. Cervical swab samples positive for HPV 16 or HPV 18, detected by real-time fluorescent quantitative polymerase chain reaction (qPCR) genotyping, were collected as the study subjects. Cervical swab samples from the same period of physical examination population that were negative for HPV 16 or HPV 18 by qPCR genotyping served as negative controls. The qRT-PCR method was employed to detect the level of miR-196a in cervical cells, with data processed via the 2^-△△Ct method.

RESULTS Differential analysis of the GSE86100 data revealed that miR-196a expression decreased in HPV 16 or HPV 18-positive cervical cells (log₂FC=-6.60, P < 0.001), while miR-3188 expression significantly increased (log₂FC=6.22, P < 0.001). Using online analysis tools https://kmplot.com/analysis, it was found that cervical cancer patients with high miR-196a expression had a shorter overall survival compared to those with low miR-196a expression (HR=1.87, 95%CI: 1.17-3.00, P=0.008). However, there was no correlation between miR-3188 and the overall survival of cervical cancer patients (HR=1.47, 95%CI: 0.92-2.37, P=0.110). The results of specific qRT-PCR testing showed that the expression levels of miR-196a in cervical cells positive for HPV 16 and HPV 18 were 0.93±0.09 and 0.51±0.07, respectively, which were lower than those in the normal control group (1.89±0.13)(P < 0.05), consistent with the sequencing analysis results.

CONCLUSIONS Infection of cervical cells with HPV 16 or HPV 18 can lead to decreased expression of miR-196a, and the expression level of miR-196a is negatively correlated with the overall survival of cervical cancer patients.