Expression Of SKA1 In Uveal Melanoma And Its Biological Significance

Objective To study the expression of SKA1(Spindle and kinetochore associated complex)in UM(Uveal Melanoma)tissues,as well as its role and molecular mechanism in the development and development of UM.It provides scientific basis for clinical accurate UM treatment with SKA1 as the target.Method 1.Detection of SKA1 expression in 20 formalin-fixed-paraffin-embedded UM tissues by immunohistochemistry,and the relationship between SKA1 and clinicopathological characteristics and survival prognosis was analyzed;meanwhile,using bioinformatics methods,TCGA(The Cancer Genome Atlas)database of UM case tissue expression of SKA1,and its relationship with prognosis and clinicopathological characteristics,supporting the results of the UM specimens of the research group.2.Use q RT-PCR(Quantitive Real Time PCR)method to detect the expression of SKA1 in three melanoma cell lines A-375,MUM-2B and MUM-2C,and select SKA1 high-expressing cell lines as tool cells,and then SKA1 was used to interfere with lentiviral infection tool cells,and nonsense sequences were used as negative control.QRT-PCR was used to detect SKA1 expression in cells of each treatment group,and effective interference sequences were screened for subsequent cell phenotypic studies.3.Using SKA1 to interfere with lentivirus-infected tool cells,using nonsense sequences as negative controls,and then using MTT method,flow cytometry,and Caspase3 / 7 method to detect proliferation and apoptosis-related proteins.To study SKA1 knockdown,tool cell proliferation Changes in the biological activity of apoptosis,clarifying the specific role of SKA1 in the occurrence and development of UM.4.Using the TCGA database UM cases,divide the SKA1 expression median into high and low expression groups,and then use GSEA(Gene set enrichment analysis)method to analyze the signal pathways that SKA1 regulates the occurrence and development of UM.Results 1.There was no significant difference in the expression of SKA1 in UM tumor tissues and adjacent tissues(P> 0.05).The results of TCGA bioassay analysis showed that the overall survival OS(overall survival)of patients with high SKA1 expression decreased significantly(P = 0.021,HR = 2.55,95% CI: 0.92-7.05).2.SKA1 was highly expressed in A-375,MUM-2B and MUM-2C melanoma cell lines.3.Compared with the negative control group(sh Ctrl),the knockdown efficiency of sh SKA1-1,sh SKA1-2,and sh SKA1-3 treatment groups were 78.86%,62.71%,and 57.83%,respectively.3.Infect MMU-2B tool cells with sh SKA1-1 interference lentivirus.Compared with the sh Ctrl group,the OD of 490 nm in the sh SKA1-1 interference group was significantly reduced(P <0.05),and the proportion of apoptotic cells in the sh SKA1-1 interference group was significantly increased.(P <0.05),the fluorescence intensity of sh SKA1-1 interference group increased significantly(P <0.05).4.GSEA analysis showed that the high expression of SKA1 was related to signal pathways related to cell cycle,meiosis,damage repair,nucleotide metabolism,DNA replication and tumorigenesis.Conclusion The results of experimental studies and biochemical analysis show that SKA1 is highly expressed in UM tissues,and its high expression is an adverse factor affecting the prognosis of UM.SKA1 may promote cell cycle transformation and inhibit apoptosis,thereby promoting the occurrence and development of UM.