Overexpression And Functional Significance Of Different LHX6 Splice Variants In Cervical Cancer

Objective: In this study,we tried to detect the expression of different LHX6 splice variants in patients with human cervical cancer and normal cervical tissue,and investigate the role of LHX6 splice variants in cervical cancer cell growth and apoptosis.Method:1.LHX6 splice variants are divided into two groups according to containing exon 12 or not.The group without exon 12 is called group LHX6-A,while the group with exon 12 is named group LHX6-B,and the group containing all the isoforms is abbreviated group LHX6-all.2.The expression of two groups LHX6-A and LHX6-B was detected by real-time quantitative PCR in m RNA level.3.Lentivirus RNA interference technology was used to inhibit the expression of group LHX6-A,LHX6-B and LHX6-all in Hela and Caski cell.4.We used Cell Counting Kit 8 analysis,EDU analysis and cell clonogenic assay to detect the proliferation of Hela and Caski cells.Apoptosis assay was used to examine apoptosis rate of cervical cancer cells.5.RNA-sequencing was used to detect the relevant genes when LHX6-B was downregulated.Results:1.LHX6-A isoforms were only be detected in cervical cancer tissue,about49.15% positive rate;in all the cervical carcinomas and normal tissues LHX6-B could be found,and was significantly upregulated in cervical carcinomas as compared to normal tissues;LHX6-B is the major component of LHX6 in cervical cancer tissue.2.Downregulation of group LHX6-B and LHX6-all both can inhibit the growth of Hela and Caski cell,induce cell apoptosis;While inhibiting the expression of group LHX6-A had no influence on the growth or apoptosis of cervical cancer cell.3.RNA-sequencing indicated that there were total 725 different genes found in group LHX6-B when compared with the control group,among which 527 genes were upregulated,and 198 genes were downregulated.Functional enrichment analysis suggested that the differential genes were involved in multiple cancer-related pathways,including the PI3K-AKT signaling pathway,Ras signaling pathway and MAPK signaling pathway.These data further suggested that transcriptome LHX6-B may exert its tumor –promoting effect by affecting several important tumor-associated pathways.Conclusion:LHX6-A isoforms and LHX6-B isoforms had different expression models in cervical epithelium,though both accumulating in cervical cancer tissues.LHX6-A can only be detected in the cervical cancer tissue,with 49.15%positive rate.But in all the cervical carcinomas and normal tissues LHX6-B could be found,and futher analysis indicates that LHX6-B is the major component of LHX6 in cervical cancer tissue.Down-regulation of LHX6 inhibited cervical cancer cell growth and induced cell apoptosis,and group LHX6-B played a major role in this process.RNA-sequencing analysis suggested that transcriptome LHX6-B may exert its tumor-promoting effect by affecting several important tumor-associated pathways,including the PI3K-AKT signaling pathway,Ras signaling pathway and MAPK signaling pathway.