The Research Of Localized-mRNA In Metastatic Liver Cancer Cells And Its Mechanism To Medicate Metastatic Property

Hepatocellular carcinoma(HCC)is the most common malignant cancer in the liver,which ranks third among the malignant cancers around the world.Nowadays,there are more and more HCC patients,approximately 250 000 people worldwide die of HCC each year.Most patients who suffer from HCC are not diagnosed at an early stage due to the fast development of this disease,which may lead to the low survival of HCC.So the early diagnosis of HCC is important for the treatment of HCC.But most of the HCC hallmarks are still at the stage of science research.A comprehensive analysis of the formation of metastasis ability is still lacking.The localization of specific mRNAs and its protein are important for the formation of metastasis ability of HCC in the cell protrusion.So it is significant to study the mRNA localization and the regulation of translation for exploring the formation of HCC metastasis.A Boyden chamber assay was used to separate the cell body fraction from the cellular protrusion fraction in both HCCLM3 cancer cells and SMMC7721 cancer cells,we combined cellular fractionation with direct RNA sequencing(DRS)to identify and quantify the total mRNA population in the cell body and protrusions of HCCLM3 cells and the SMMC-7721 cells.We screen out the high-expressed mRNA in the protrusion of highly metastatic HCCLM3 cancer cells and then made an analysis of the specific mRNA and its protein by RT-qPCR,RNA FISH,Western blotting and immunofluorescence,si-RNA targeted to specific mRNA was conducted and then proliferation,migration assays and in vitro invasion assay were utilized to detect the influence of specific localized-mRNA to the ability of HCCLM3 cancer cells.we found that the the metastasis ability of HCCLM3 decreases after stat3 was knockdown,which suggests that stat3 mRNA may play an important role in the process of the formation of metastasis ability.Besides,we used co-immunoprecipitation to test the interaction between stat3 and platelet-derived growth factor receptor?vascular endothelial growth factor receptor?hepatocyte growth factor receptor and the non-hepatic related nerve growth factor receptor.we found that the non-phosphorylated Stat3 in cell protrusion has interaction with PDGFR,and this process is relied on growth factor.the result reveals the mechanism of the metastasis ability regulation process via localized-mRNA.