Correlation Study Between Differential Expression Level Of TRNAs And Mesothelioma Immunophenotypes

ObjectiveMesothelioma is a kind of malignant tumor.Although the incidence rate is low,it is on the rise,and the mortality is high.The median survival time is only 10-12 months.It is speculated in the literature that China will face a new peak of mesothelioma like other countries.Due to the long incubation period of mesothelioma for 30-40 years,the lack of specific clinical symptoms and the lack of specific detection indicators,the research lag of mesothelioma hinders the improvement of the diagnosis and treatment of mesothelioma.TransportRNA（tRNA）has tissue specificity and relative stability.The expression level of tRNA is related to the occurrence and development of breast cancer,prostate cancer and other tumors,but the correlation with mesothelioma has not been reported.This topic uses high-throughput sequencing method to screen travel specific tRNA,studies the relationship between common immunophenotypes and tRNA expression level in clinicopathological diagnosis of mesothelioma,and expounds whether the differential expression level of tRNA may affect the expression of immunophenotype related proteins,so as to provide tRNA level scientific data for the difference of immunophenotype of mesothelioma.Methods1.The selected samples were paraffin embedded mesothelioma tissues,from 60 paraffin specimens diagnosed as mesothelioma by the pathology department of Liaoning Cancer Hospital from 2003 to 2019.The expressions of CK5/6,WT1,CR,CK,CK7,MC,D2-40,Vim and S-100 were detected by immunohistochemical staining,and the immunophenotypic indexes with high sensitivity and specificity were screened.2.Taking 60 cases of mesothelioma as the research background,6 samples with significant differences in immunophenotypic sensitivity and specificity were selected and divided into the experimental group with high sensitivity and specificity and the control group with low sensitivity and specificity for high-throughput tRNA sequencing.The significant tRNAs between the two groups were compared and verified by PCR.3.Bioinformatics predicts whether the target genes that may be regulated by the screened differential tRNAs have correlation and preference with the immunophenotype of mesothelioma.Results1.In 60 cases of mesothelioma,pleural mesothelioma was mainly male（54.1%）and peritoneal mesothelioma was mainly female（87.0%）;Pleural mesothelioma under50 years old was less than peritoneal mesothelioma（40.9%）,and peritoneal mesothelioma over 50 years old was more than pleural mesothelioma（63.2%）;Pleural mesothelioma was mainly less than 2.0 cm in diameter（55.6%）,and peritoneal mesothelioma was more than 2.0 cm in diameter（69.0%）.The overall survival rate of male mesothelioma patients（30.6%）was lower than that of female mesothelioma patients（46.4%）,and that of pleural mesothelioma patients（30.2%）was lower than that of peritoneal mesothelioma patients（38.8%）.There was no trend relationship between histological type,age and tumor diameter and the overall survival rate of mesothelioma patients.Immunohistochemical staining of paraffin tissue of 60 cases of mesothelioma showed that the positive expression rates of CK5/6,WT1 and CR in epithelioid mesothelioma were higher than those in sarcomatoid mesothelioma.2.According to the difference of sensitivity and specificity of immunophenotype,6 cases of mesothelioma were selected,including 4 cases of epithelioid mesothelioma,1case of sarcomatoid mesothelioma and 1 case of biphasic mesothelioma.Nine differentially expressed tRNAs were screened by tRNA high-throughput sequencing.tRNA-Asn-GTT-10-1 and tRNA-Gln-TTG-2-1 were verified by real-time quantitative PCR;SATL1,H1F0,PRB2,SMCP,RPL29,SEMG1,SEMG2,LYRM2 and PRB1 may be related to the differential expression of immunophenotype of mesothelioma in this group.Conclusion1.In men,mesothelioma patients with tumor diameter <2.0cm occurred more in the pleura than in the peritoneum,and in women,mesothelioma patients with tumor diameter ≥2.0cm occurred more in the peritoneum than in the pleura.The overall survival rate of male mesothelioma patients was lower than that of female mesothelioma patients,the overall survival rate of patients with pleural mesothelioma was lower than that of patients with peritoneal mesothelioma.The sensitivity and specificity of CK5/6,WT1 and CR were significant.2.In the two groups of cases with differential expression of CK5/6,WT1 and CR,tRNA high-throughput sequencing found 9 differentially expressed tRNAs,and bioinformatics predicted 218 genes associated with differentially expressed tRNAs.3.Real-time quantitative PCR showed that tRNA-Asn-GTT-10-1 and tRNA-GlnTTG-2-1 were the most significantly different tRNAs;SATL1,H1F0,PRB2,SMCP,RPL29,SEMG1,SEMG2,LYRM2 and PRB1 may be related to the differential expression of immunophenotype of mesothelioma in this group.