Study On The Mechanism Of Action Of Lonp1 Protein In Esophageal Cancer

Objectives To detect the expression of Lonp1 protein in esophageal carcinoma tissues and cells,and to study the effect of Lonp1 protein on the malignant phenotypes of invasion,migration and proliferation of esophageal cancer cells,and further explore the mechanism of action of Lonp1 protein in esophageal cancer.Methods The Lonp1 protein was searched using a biological information library for bioinformatics analysis.Immunohistochemical staining was used to detect the expression of Lonp1 in cancer and adjacent cancer in 75 tissue microarrays(Shanghai Core Super Biotechnology Co.,Ltd.)paired with esophageal cancer.Western blot was used to detect the expression of Lonp1 protein in esophageal cancer cells,and the expression of Lonp1 protein in different esophageal cancer cell lines was detected.Primer 5.0 software designed primers for the Lonp1 protein for primer-specific validation.The expression level of Lonp1 mRNA in esophageal cancer cell lines was detected by RT-PCR.RNA extraction of esophageal cancer tissues was performed,and the expression levels of cancer and adjacent Lonp1 protein were detected by RT-PCR.Cell lines YES2 and KYSE150 with high expression of Lonp1 protein were selected and knockdown experiments were performed using RNAi technology.Transwell experiments and colony formation assays were performed to examine the effect of knockdown of Lonp1 protein on the invasion,migration and proliferation of esophageal cancer cells.Overexpression of Lonp1 protein cell lines KYSE30 and KYSE450 was performed using plasmid transfection for overexpression experiments.Transwell assay and colony formation assay were used to detect the effect of overexpressing Lonp1 protein on invasion,migration and proliferation of esophageal cancer cells.Western blot was used to detect the effect of Lonp1 protein on PI3K/AKT pathway and EMT pathway.Screening of natural small molecule drugs revealed that natural tea extract epigallocatechin gallate not only inhibited esophageal cancer cells,but also affected Lonp1 protein.Results The percentage of Lonp1 positive expression in 75 ESCC samples was 88%(66/75),and the percentage of Lonp1 negative was 12%(9/75)in esophageal cancer tissue microarray.The expression of Lonp1 protein was significantly higher than that of cancer.The expression of Lonp1 protein in esophageal cancer cell lines was significantly higher than that in immortalized esophageal epithelial cell lines.The expression levels of Lonp1 protein in YES2 and KYSE150 cell lines after RNA interference treatment were significantly lower than those in control NC group.At the same time,compared with the NC control group,the invasion,migration and colony formation ability of the YES2 and KYSE150 cells in the knockdown group were significantly decreased(P<0.05).The expression of post-Lonp1 protein in plasmid overexpressing KYSE30 and KYSE450 strains was significantly higher than that in the empty group.At the same time,compared with the empty control group,the proliferation,migration and colony forming ability of KYSE30 and KYSE450 cells in the overexpression group were significantly increased(P<0.05).The expression of AKT protein in the esophageal cancer cells YES2 and KYSE150 knockdown Lonp1 protein,PI3 K,and phosphorylated Ser473 was significantly decreased.Overexpression of Lonp1 protein,N-cadherin,Vimentin protein expression and E-cadherin protein expression decreased in esophageal cancer cells KYSE30 and KYSE450.It was found by small molecule drug screening that the concentration of epigallocatechin gallate(EGCG)in the natural tea extract increased the expression of Lonp1 in the esophageal cancer cell line KYSE150.Conclusions Lonp1 protein is highly expressed in esophageal cancer tissues and cell lines.The expression of Lonp1 protein is significantly correlated with TNM stage and lymphatic metastasis.Knockdown of Lonp1 protein can effectively inhibit the invasion,migration and colony forming ability of esophageal cancer cells.Overexpression of Lonp1 protein can significantly enhance the invasion,migration and colony forming ability of esophageal cancer cells.Knockdown of Lonp1 protein inhibits the PI3K/AKT signaling pathway.Overexpression of Lonp1 protein promotes the esophageal cancer epithelial-mesenchymal transition(EMT)pathway.Epigallocatechin gallate inhibits proliferation of esophageal carcinoma and induces apoptosis.The amount of Lonp1 protein expression gradually decreased as the concentration of epigallocatechin gallate(EGCG)increased.Figure 15;Table 2;Fefernces 60...