Bioinformatics Analysis Of The Role Of LRP11 In Hepatocellular Carcinoma And Its Effect On The Biological Behavior Of Hepatoma Cells

Background:Hepatocellular carcinoma(HCC)is the main pathological type of primary liver cancer.As a multifactorial disease associated with tumor microenvironment,immune evasion,and immune tolerance,its mortality rate has been high,therefore,finding some effective biomarkers and therapeutic targets can help improve the prognosis survival of patients.Low-density lipoprotein receptor-related protein 11(LRP11)is a cell membrane receptor protein that belongs to the LRP family.The LRP family plays an important biological role in lipid metabolism,and several members of the family have been linked to cancer metastasis,transformation,and progression.Lipids with biological activity are mediators of the carcinogenesis process.LRP11 has been shown in studies to play an oncogenic role in the development and progression of multiple cancers and to have good prognostic value.The role and mechanism of LRP11 in HCC,however,remain unknown.Objective:This project’s objective is to research the expression level of LRP11 in HCC patients and its correlation with survival prognosis.And to conduct experiments to determine the effect of LRP11 on the proliferation and migration of hepatoma cell lines.Methods:The expression levels of LRP11 in pan-cancer were analyzed using the TIMER online database.To understand the expression level and prognosis of LRP11 in HCC patients,HCC transcriptomic and clinical data were downloaded from three databases: TCGA,ICGC,and GEO.TCGA clinical data was used to create Cox regression risk models,and to investigate the relationship between LRP11 and clinicopathological parameters.Kaplan-Meier Plotter online database analysis of the effect of LRP11 on different survival status periods of patients.Twenty pairs of HCC tissues were gathered from the Second People’s Hospital of Changzhou affiliated to Nanjing Medical University,and the differential expression of LRP11 in tumor and paracancerous tissues was determined using quantitative real-time polymerase chain reaction(qRT-PCR).The protein expression of LRP11 in hepatoma cell lines was detected using Western blot(WB).The effects of LRP11 on the proliferation of hepatoma cell lines were studied by CCK8 assay,Colony-Formation assays,and EDU assay.Transwell assay was used to study the effect of LRP11 on hepatoma cell line migration.Using the TIMER online database,we investigated the relationship between LRP11 and tumor immune cell infiltration further.Gene Set Enrichment Analysis(GSEA)was used to investigate potential biological signaling pathways linked to LRP11 in HCC.Results:1.Analysis of the database showed that LRP11 was highly expressed in a variety of cancers.In the TCGA,ICGC,and GEO databases,we also discovered that LRP11 was highly expressed in HCC(TCGA: P<2.22e-16;ICGC: P<2.22e-16;GSE45267: P=4e-13;GSE54236: P=0.00066;GSE62232: P=0.00037;GSE64041: P=1.8e-07),Moreover,high expression of LRP11 was associated with poor prognosis in patients with HCC(TCGA: P=0.041;ICGC: P=0.021),a result further confirmed in the Kaplan-Meier Plotter online database,and high expression of LRP11 affects the different survival status of HCC patients,including disease specific survival(P = 0.0071),overall survival(P = 0.0095),progression free survival(P = 0.00031)and recurrence free survival(P = 0.0019).Univariate and multivariate regression analyses showed that LRP11 could be an independent prognostic factor for HCC patients.2.qRT-PCR analysis demonstrated that LRP11 expression is significantly higher in HCC tissues compared to paracancerous non-tumor tissues.WB showed that LRP11 protein abundance was higher in hepatoma cell lines than in normal hepatocytes.In addition,knockdown of LRP11 inhibited hepatoma cell lines proliferation and migration.3.GSEA indicates that LRP11 is involved in multiple cancer-related signaling pathways.LRP11 was significantly correlated with a variety of immune infiltrating cells,which were associated with B cells(Partial cor=0.272,P=2.89e-07),CD8+T cells(Partial cor=0.199,P=2.14e-04),CD4+T cells(Partial cor=0.348,P=3.12e-11),Macrophages(Partial cor=0.368,P=2.11e-12),Neutrophils(Partial cor=0.371,P=1.03e-12)and Dendritic cells(Partial cor=0.322,P=1.26e-09)were positively correlated.In addition,LRP11 was also positively correlated with several immune checkpoint markers,including Lymphocyte activation gene 3(LAG3)、CytotoxicT-lymphocyte protein 4(CTLA4)、Programmed Cell Death Protein 1(PDCD1)and T cell immunoglobulin and ITIM domain(TIGIT).Conclusion:1.In HCC,high expression of LRP11 was positively correlated with poor prognosis of patients and could serve as an independent prognostic factor with the potential to become a prognostic marker for HCC.2.LRP11 may be involved in the development and progression of HCC,promoting the growth and migration of hepatoma cell lines in vitro.3.LRP11 was positively correlated with immune cell infiltration and immune checkpoint markers,thus LRP11 could be a potential target for HCC immunotherapy.