SNORA71A通过TLR3/PD-L1通路调控食管鳞状细胞癌TE-1细胞的恶性生物学行为OA

Objective:To investigate the effects of small nucleolar RNA SNORA71A on the proliferation,migration,and invasion of esophageal squamous cell carcinoma(ESCC)cells via TLR3/PD-L1 expression.Methods:qPCR was used to detect SNORA71A expression in tumor tissues,adjacent non-tumor tissues from 52 ESCC patients and human ESCC cell lines.Human ESCC TE-1 cells were transfected with antisense oligonucleotides(SNORA71A-ASO-29,SNORA71A-ASO-102,NC-ASO)or small interfering RNAs(siTLR3,siTLR3-NC),designated as SNORA71A-ASO1 group,SNORA71A-ASO2 group,NC-ASO group,and siTLR3 group,respectively.Additionally,the overexpression plasmid pcDNA3.1SNORA71A and the empty vector pcDNA3.1 were transfected into TE-1 cells,referred to as the SNORA71A group and Vector group,respectively.Cell proliferation,migration,and invasion capabilities after SNORA71A knockdown or overexpression were evaluated using cell functional assays,namely MTS,scratch wound-healing,and Transwell invasion assays.High-throughput transcriptome sequencing was performed to identify downstream target genes of SNORA71A,and Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)functional enrichment analyses were employed to predict associated biological processes and signaling pathways.qPCR was used to detect the expression of the downstream target gene TLR3 in ESCC tissues and cells.Additionally,the mRNA and protein expression levels of PD-L1 in TE-1 cells after TLR3 knockdown or overexpression were measured using qPCR and WB.Cell functional assays were performed to examine the impact of TLR3 knockdown on the malignant biological behaviors(proliferation,migration,and invasion)of TE-1 cells promoted by SNORA71A.Results:SNORA71A expression was significantly upregulated in tumor tissues from 52 ESCC patients and in ESCC cell lines(P＜0.01 or P＜0.05).Knockdown of SNORA71A inhibited TE-1 cell proliferation,migration,and invasion(P＜0.01 or P＜0.05),whereas SNORA71A overexpression promoted these malignant behaviors(P＜0.01 or P＜0.05).High-throughput transcriptome sequencing identified TLR3 as a downstream target gene of SNORA71A.TLR3 expression was significantly downregulated in ESCC tissues and TE-1 cells(P＜0.01 or P＜0.05).In addition,TLR3 positively regulated the mRNA and protein expression of PD-L1(P＜0.01 or P＜0.05).In the cell functional assays,TLR3 partially attenuated the regulatory effect of SNORA71 Aon PD-L1 expression(P＜0.01).Conclusion:SNORA71A regulates the proliferation,migration and invasion of TE-1 cells by modulating the TLR3/PD-L1 pathway.