The Role Of LAMP2A In The Development And Sensitivity To Cisplatin Of Esophageal Squamous Cell Carcinoma

Backgrounds and aimsEsophageal cancer is one of the common malignancies.Esophageal squamous cell carcinoma(ESCC)accounts for about 90%of the total number of esophageal cancer in China.High incidence,high mortality,poor prognosis,lacking of effective clinical treatment and other reasons made esophageal cancer widely concerned.Deeper investigation and new treatment strategies are urgently needed.Lysosome-associated membrane glycoprotein 2 isoform A precursor(LAMP2A),one of the key proteins in chaperone-mediated autophagy,has been reported to be involved in the genesis and development of multiple tumors,but its role in ESCC has not been clarefied.The aim of this study was to investigate the effect of LAMP2A on malignant traits of ESCC cells and to determine the role of LAMP2A in the development of ESCC,providing a new idea for targeted therapy of ESCC.MethodsIn this study,the expression of LAMP2 in esophageal cancer tissues was determined by bioinformatics methods in the TCGA database and GTEx database.LAMP2A knockdown or overexpressed ESCC cell strains were established using a second-generation lentiviral packaging system(three-plasmids system).Positive cells were selected using puromycin,and then Real-time PCR and Western Blotting were used to determine the mRNA and protein expression of LAMP2A in these ESCC cell strains.Secondly,the modified ESCC cell strains were cultured in vitro,and the effects of LAMP2A on cell proliferation,migration and colony formation ability were investigated.Cell proliferation was dynamically monitored in the IncuCyte Zoom,and the effect of LAMP2A on cell migration was examined by wound scratch assay.Plate colony formation assay was used to dectect the role of LAMP2A in the colony formation ability of ECSS cells.Meanwhile,the modified ESCC cells were treated with cisplatin in this study,and cell viability was measured by MTS assay to determine the effect of LAMP2A on sensitivity of ESCC cells to the drug.Results1.The expression of LAMP2 is higher in esophageal cancer tissues than that in normal esophageal tissues,and that patients with higher LAMP2 expression have shorter overall survival curves.2.Stable ESCC cell strains with LAMP2A-modified were constructed.The expression of LAMP2A in KYSE150 and KYSE510 modified by pLKO-shLAMP2A(Lentiviral particles to knockdown LAMP2A)was significantly decreased,determined by Real-time PCR and Western Blotting.And the mRNA expression of LAMP2A in KYSE410 and KYSE180 modified by FUB-LAMP2A(Lentiviral particles to overexpress LAMP2A)was significantly increased.3.Cell proliferation,cell migration and colony formation ability were reduced in LAMP2A-knockdown ESCC cells.In contrast,these malignant traits were promoted in LAMP2A-overexpressed ESCC cells.4.Sensitivity to cisplatin was increased in LAMP2A-knockdown cells.Conclusion1.LAMP2A promotes cell proliferation,migration and colony formation ability of ESCC cell lines.2.LAMP2A protects ESCC cells from cisplatin treatment.