Correlation And Mechanism Between PI3K Signaling Pathway MiRNA Gene Polymorphisms And Susceptibility To Cervical Cancer

Background:Cervical cancer is the fourth leading cause of cancer death among women worldwide.Persistent infection with high-risk HPV is the most important risk factor for cervical cancer,and genetic factors also play a role in individual cervical cancer susceptibility differences.The study found that SNPs in miRNA genes of PI3K signaling pathway may play a role in the pathogenesis process by influencing the expression of miRNAs and their interaction with target genes.Therefore,studying the correlation between SNPs in miRNA genes and the risk of cervical cancer and its mechanism will help elucidate the pathogenesis of cervical cancer and new molecular targets for cervical cancer diagnosis and treatment.Methods:Correlation between PI3K signaling pathway miRNA gene polymorphisms and cervical cancer risk:firstly,abnormally expressed miRNAs in cervical cancer were screened;The target genes of these miRNAs were selected through the TargetScan database;The miRNA target genes were enriched through the DAVID database,and the miRNA genes related to the PI3K signaling pathway were screened.Using Ensemble and other databases and screened miRNA gene-related SNPs(rs 107822 for miR-219a,rs10877887 for let-7i,rs2292832 formiR-149,rs353293 for miR-143,rs3746444 for miR499,rs3803808 for miR-132,rs4078756 for miR-10b,rs629367 for let-7a,rs7372209 for miR-26a);A total of 431 patients with cervical cancer,273 patients with precancerous lesions of cervical cancer,and 698 healthy controls were randomly selected,The MassARRAY nucleic acid mass spectrometry was used to genotype the SNP loci in the population.The correlation between SNP alleles and genotypes in miRNA genes and the risk of cervical cancer precancerous lesions and cervical cancer was analyzed.Mechanism of PI3K signaling pathway miRNA gene polymorphisms’ correlation with cervical cancer risk:Based on the research results of the previous research by our group,the miRNA gene miR-196a-3p associated with PI3K signaling pathway was screened.The expression level of miR-196a-3p and its target gene in cervical cancer tissues and adjacent normal tissues was analyzed by qPCR and Steam-loop RT-qPCR,respectively.The interaction of miR196a-3p(mimic or mimic carrying different alleles of rs11614913)with target genes was studied by the double luciferase reporter detection system;The effect of rs11614913 on the expression of miR-196a-3p and target gene was studied by constructing overexpression vectors of miR-196a-3p carrying different alleles of rs11614913.The proliferation ability of cervical cancer cells overexpressing or knocking out miR-196a-3p or target genes was detected by CCK8 experiment;Flow cytometry was used to detect the apoptosis ability of cervical cancer cells that overexpressed or knocked out the miR-196a-3p target gene.Result:Correlation between miRNA gene polymorphisms in PI3K signaling pathway and cervical cancer risk:The difference between the allele frequencies of the SNP locus rs107822 in the miR-219 gene and the SNP locus rs2292832 in the miR-149 gene was statistically significant between the cervical cancer group and the control group(P=0.004 and P=0.004).Among them,the rs107822 locus allele C was a risk factor for cervical cancer(OR=1.292,95%CI:1.088-1.536).Allele C at the rs2292832 locus was a protective factor for cervical cancer(OR=0.767,95%Cl:0.639-0.921).The results of genetic pattern analysis showed that the difference in the distribution frequency of genotypes of rs107822 and rs2292832 loci between cervical cancer group and control group was statistically significant(P=4.5x10-3,P=0.004),and the optimal inheritance pattern of both loci was Log-addictive,in which the 2CC-CT genotype of rs107822 locus may be a risk factor for cervical cancer(OR=1.28,95%CI).:1.08-1.51);The 2CC-CT genotype at the rs2292832 locus may be a protective factor for cervical cancer development(OR=0.76,95%CI:0.640.92).The mechanism of PI3K signaling pathway miRNA gene polymorphisms correlated with the risk of cervical cancer:the expression level of miR-196a-3p in cervical cancer tissues was significantly higher than that in adjacent normal tissues.miR-196a-3p can significantly promote the proliferative ability of cervical cancer cells;miR-196a-3p plays a role in cervical cancer cells by targeting CASP9 in the PI3K signaling pathway;CASP9 can inhibit the proliferation ability of cervical cancer cells and enhance the apoptosis ability of cervical cancer cells;In cervical cancer cells,allele C at the rs11614913 locus was associated with higher miR-196a-3p expression levels and lower CASP9 expression levels than allele T.The allele C at the rs11614913 locus is more conducive to the negative regulation of CASP9 by miR-196a-3p than the allele T.Conclusion:The SNP locus rs107822 in the miR-219 gene and the SNP locus rs2292832 in the miR-149 gene are associated with cervical cancer susceptibility.miR-196a-3p plays the role of oncogene in cervical cancer by targeting CASP9,and the SNP locus rs11614913 in this miRNA gene can affect cervical cancer development by influencing the expression of miR-196a-3p and the interaction of miR-196a-3p with the target gene CASP9.