Study On The Induction Of Malignant Progression Of Uveal Melanoma And Glioma By Autophagy Adaptor Protein P62/SQSTM1 Based On Proteomics Analysis

BACKGROUND&OBJECTIVE:With the deepening of cancer research,the understanding of the relationship between tumor cells(TC)and the tumor microecological environment(TME)where the tumor is located is constantly deepening.In the initial stage,TCs are attacked by parasitic immune cells and reconstruct parasitic tissues.This reconstruction actually involves the transformation of cells that attack TCs.Once the transformation is successful,TCs not only survive,but also gain the potential for malignant progression.The molecular mechanism for TCs to achieve malignant progression in TME has not been fully elucidated,especially the target molecules that play a key role in this process still need to be further revealed.In this study,proteomic analysis was designed to screen key proteins in the progression of TME-induced TCs malignancy.The aim is to provide experimental basis for the study of molecules that can target both TC and TME bidirectional therapy.MATERIALS AND METHODS:?Uveal melanoma 92.1 cells post-ocular inoculation in nude mice,the preparation of the original generation of single cell culture in vitro after the tumorigenic,monoclonal culture the high proliferation force cell,related molecular biology testing and equipment are carried.?The full quantitative analysis of iTRAQ/TMT protein and the target protein screening and validation were carried.?Based on the proteomies analysis,screening the key gene related to the key functional protein from TCGA,validating the gene in clinical surgical specimens and cell lines of gliomaRESULTS AND ANALYSIS:? 92.1 cells were inoculated after the eyeball,and grew in the adjacent tissues of uveal of the recipient animal in the early stage.The mid-term was confined to the orbit,late to the extermal orbital region,intracranial,lung and kidney,approximately reaching the grape of requirements for in situ vaccination model of melanoma.?A high proliferative cell(92.1B)was successfully cloned from the transplantation tumor tissue,and the tumor biological characteristics of this cell were confirmed to be more malignant than the original 92.1 by relevant molecula biological detection and tumoregenic experiment.?From 92.1,92.IB and 92.1 A(derived from the monoclonal reculture of 92.1 nude mouse ventricular xenograft tumor,were confirmed more malignant than 92.1),SQSTM1/P62,CDK1 and CASP3 were screened from the full-quantity analysis of iTRAQ/TMT proteins.Bioinformatics analysis suggested that the three proteins were the key proteins regulating 92.1 malignant progression.After further experimental verification,it was found that P62/SQSTM1 was correlated with the induction of up-regulation of gene expression of multipotent stem cells such as Nanog,SOX2,OCT4,and the expression of GBP1.P62/SQSTM1 was related to the prognosis of clinical patients.CONCLUTIONS AND PROPECTS:? Human uveal melanoma heterogeneous orthotopic transplantation animal model is superior to the pure animal model(mouse melanoma cells transplantation tumor model in rat or rabbit)However,eyes of immunodeficient mice which were used as receptors are too small,there are certain difficulties in production.In the fiele of uveal melanoma research,we report for the firt time to establish spontaneous green fluorescent protein recpeter animal moded to trace TME.However,the cells used in the experiments are cell lines that have been grown in vitro for a long time.In order to avoid the potential molecular wariation caused by long-term transmission in vitro,it is proposed to produce a xenograft animal model derived from the clinical fresh human uveal melanoma surgical specimens.?The full quantitative analysis of iTRAQ/TMT protein for cancer molecula mechanism research is becoming more and more practical and rapid,and this article first report in TC and TME correlation study.P62/SQSTM1,CDK1 and CASP3 are the key proteins to TME-induced malignant progression of TC.Is there a causal relationship between P62/SQSTM1,CDK1 and CASP3 in the regulation of TME-induced malignant progression of TC,and whether TC appears benign transformation after knocking down expression or using inhibitors?It remains futher study.?GBP1 promotes tumor cell invasion in glioma.This study first reported the correlation between the expression of GBP1 and the malignant progression of glioma and the prognosis of clinical patients.The role of P62/SQSTM1 as a key protein in the GBP1 pathway is worth further study.