Study On Talin-1 Interaction Network Promoting The Growth And Metastasis Of Hepatocellular Carcinoma

Background and aim: Talin-1,a macromolecules cytoskeletal protein,could promote the process of hepatocellular carcinoma(HCC).However the related mechanism remains unclear.In this study,Whole Human Genome Oligo Microarray with verification using HCC cells and sh-Talin-1 HCC cells was performed to analysis the mechanism of Talin-1 in promoting HCC progression.Method: Real-time quantitative reverse transcription-polymerase chain reaction(q RT-PCR)and Western blot(WB)were respectively used to detect the expression of Talin-1 m RNA and protein in hepatocellular carcinoma cell line MHCC-97 L and normal liver epithelial cell LO2.Lentiviral vectors were used for MHCC-97 L cell transduction to establish a stable Talin-1 knockdown MHCC-97 L cell line(sh-Talin-1 MHCC-97L).Cell counting method was used to analyze the growth capacity of MHCC-97 L cells and sh-Talin-1 MHCC-97 L cells.Cell scratching and Transwell migration assays were performed to detect the transfer ability of MHCC-97 L cells and sh-Talin-1 MHCC-97 L cells.Human Genome Oligo Microarray(40000 genes)with verification using HCC cells and sh-Talin-1 HCC cells and Gene Ontology(GO)project using the microarray data were performed to analysis the genes and biological behaviors regulated by Talin-1.Cytoscape v3.4.0 was used to perform the interaction networks of Talin-1.Graph Pad Prism 5 was used for statistical analysis.Result: Talin-1 was expressed in both MHCC-97 L and LO2 cells,and the expreesion level of both Talin-1 m RNA and protein were higher in MHCC-97 L than those in LO2 cells.In addition,the expreesion level of both Talin-1 m RNA and protein were higher in MHCC-97 L than those in sh-Talin-1 MHCC-97 L cells.Via Whole Human Genome Oligo Microarray,we found that of the 40,000 tested genes,3099 were differentially expressed after Talin-1 knockdown;expression of 1924 genes was increased,while expression of 2175 was decreased.Through GO profile,we found Talin-1 promotes many HCC advanced related biology behaviors,including ion transport(related genes: CAV3,KCNE3),membrane depolarization(related genes: CAV3,KCNE3),cell growth(related genes: EDN1,IGFBP5,CYR61)and cell adhesion(related genes: MUC1,SLAMF7,WISP1).We also characterized the network of gene transcripts regulated by Talin-1 and their role in promoting HCC progression.Finally,through searching published reports and online bioinformatic databases,we found that Talin-1 can regulate cell cycle genes(Cyclin D1,BCL-2 family and p53 network),EMT related genes(E-cadherin,N-cadherin and vimentin),PI3K/AKt/m TOR pathway and so on.Conclusion: 1,Talin-1 was highly expressed in MHCC-97 L cells compared with normal liver epithelial cells.2,Talin-1 can promote the growth and metastasis of MHCC-97 L cells.3,Talin-1 can promote many biological behavior involved in HCC progression.4,Talin-1,through a complex regulatory network,promote the growth and metastasis of MHCC-97 L cells.