图1 MIS18BP1在泛癌及肾透明细胞癌中的表达特征

Fig.1 Expression patterns of MIS18BP1 in pan-cancer and renal clear cell carcinoma. A: Differential expression of MIS18BP1 in pan-cancer. *P<0.05, **P<0.01, ***P<0.001 vs Normal. B: Differential expression of MIS18BP1 in KIRC. ****P<0.0001. C: Paired difference analysis. ****P<0.0001. D: Expression of MIS18BP1 protein in renal clear cell carcinoma tissue and adjacent non-cancerous tissue (Original magnification: ×200). *P<0.05 vs adjacent tissues, #P<0.05 vs clinical stage I-II. E: Expression of MIS18BP1 protein in normal renal epithelial cells and renal cancer cell lines. *P<0.05 vs HK-2.

图2 MIS18BP1对KIRC的诊断价值

Fig.2 Diagnostic value of MIS18BP1 for KIRC. A: MIS18BP1 ROC curve for predicting overall survival in TCGA. B: ROC curve of MIS18BP1 for predicting overall survival in GSE36895. C: ROC curve of MIS18BP1 for predicting overall survival in GSE53757. D: ROC curve of MIS18BP1 for predicting overall survival in GSE66272.

图3 MIS18BP1表达与KIRC临床病理特征及免疫浸润的关系

Fig.3 Correlation of MIS18BP1 expression level with clinical pathological features and immune infiltration in KIRC. A: Relationship between MIS18BP1 expression and T stage. B: Relationship between MIS18BP1 expression and N stage. C: Relationship between MIS18BP1 expression and M stage. D: Relationship between MIS18BP1 expression and grade. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001. E: Relationship between MIS18BP1 expression and immune cells. F: Differential expression of MIS18BP1 in immune cells. *P<0.05.

图4 WGCNA识别MIS18BP1相关基因模块

Fig.4 Weighted gene co-expression network analysis (WGCNA) for identifying MIS18BP1-related gene modules. A: Sample clustering dendrogram. B: Determination of the optimal soft threshold (left: scale independence assessment; right: average connectivity analysis), with the red reference line indicating R²=0.9. C: Gene clustering dendrogram (branches represent individual genes, and the colored blocks at the bottom indicate co-expression modules). D: Module-trait association heatmap (colored blocks represent co-expression modules, red and blue indicate positive and negative correlations with correlation coefficients and P-value annotations).

图5 MIS18BP1相关基因的筛选及其功能通路富集分析

Fig.5 Screening of MIS18BP1-related genes and their GO/KEGG analysis. A: Screening key genes related to MIS18BP1 based on the PPI network. Node size is proportional to degree centrality, and node color intensity is proportional to betweenness centrality (dark red indicates high betweenness). B: Correlation analysis between key genes and MIS18BP1 expression. *P<0.05, ***P<0.001. C: GO and KEGG enrichment analysis of key genes.

图6 敲低肾癌细胞中的MIS18BP1

Fig.6 Knockdown of MIS18BP1 in renal cancer cells. A: Western blotting for detection of MIS18BP1 protein expression in 786-O cell line. B: Western blotting for detection of MIS18BP1 protein expression in ACHN cell line. *P<0.05 vs sh-MIS18BP1.

图7 敲低MIS18BP1蛋白抑制肾癌细胞的增殖能力

Fig.7 Knocking down MIS18BP1 inhibits proliferation of renal cancer cells. A, C, E: Knockdown of MIS18BP1 significantly inhibits proliferation of 786-O cells (Scale bar=200 μm). B, D, F: Knockdown of MIS18BP1 significantly inhibits proliferation of ACHN cells (Scale bar=200 μm). *P<0.05, **P<0.01, ****P<0.0001 vs Vector.

图8 敲低MIS18BP1蛋白抑制肾癌细胞的迁移能力

Fig.8 Knocking down MIS18BP1 inhibits migration ability of renal cancer cells. A: Knocking down MIS18BP1 in inhibits migration ability of 786-O cells (Scale bar=500 μm). B: Knocking down MIS18BP1 inhibits migration ability of ACHN cells (Scale bar=500 μm). *P<0.05, **P<0.01 vs Vector.

图9 MIS18BP1对肾癌细胞EMT相关蛋白表达的影响

Fig.9 Effect of MIS18BP1 on expression of EMT-related proteins in renal cancer cells. A: MIS18BP1 regulates the expression of EMT-related proteins in 786-O cells. B: MIS18BP1 regulates the expression of EMT-related proteins in ACHN cells. *P<0.05, **P<0.01 vs Vector.

图10 MIS18BP1下调对PI3K/AKT信号通路的影响

Fig.10 Effect of MIS18BP1 knockdown on the PI3K/AKT signaling pathway. A: MIS18BP1 regulates the expression of key proteins in the PI3K/AKT signaling pathway in 786-O cells. B: MIS18BP1 regulates the expression of key proteins in the PI3K/AKT signaling pathway in ACHN cells. *P<0.05, **P<0.01 vs Vector.

图11 敲低MIS18BP1抑制786-O和ACHN细胞增殖及迁移,该作用可被740Y-P部分逆转

Fig.11 Knockdown of MIS18BP1 inhibits proliferation and migration of 786-O and ACHN cells, which is partially reversed by 740Y P. A, B: CCK-8 assay for assessing proliferation activity of 786-O (A) and ACHN (B) cells. C, D: Wound healing assay for assessing migration ability of 786-O (C) and ACHN (D) cells (Scale bar=200 μm). E, F: Transwell assay for detecting 786-O (E) and ACHN (F) cells passing through the membrane (Scale bar=500 μm). *P<0.05, **P<0.01 vs sh-MIS18BP1 (C-F).