MIS18BP1 regulates epithelial-mesenchymal transition, proliferation, and migration of renal cancer cells via the PI3K/AKT pathway

doi:10.12122/j.issn.1673-4254.2026.09.15

Abstract

Abstract:

Objective To explore the regulatory role of MIS18BP1 in epithelial-mesenchymal transition (EMT), proliferation, and migration of renal cancer cells and its molecular mechanism. Methods TCGA and GEO databases were used to analyze the expression patterns of MIS18BP1 in kidney renal clear cell carcinoma (KIRC), and its differential expression between tumor and adjacent tissues was validated by immunohistochemistry. The diagnostic value of MIS18BP1 and its correlation with clinicopathological parameters and immune infiltration were evaluated. Weighted gene co-expression network analysis (WGCNA) was performed to identify MIS18BP1-related gene modules for functional enrichment analysis. In renal cancer cell lines (786-O and ACHN), the effects of lentivirus-mediated MIS18BP1 knockdown on cell proliferation and migration were evaluated using CCK-8, colony formation, Transwell, and wound healing assays. Western blotting was performed to determine the expression levels of EMT-related proteins and PI3K/AKT pathway-related proteins, and rescue experiments were carried out using 740Y-P (a PI3K activator). Results MIS18BP1 was significantly overexpressed in KIRC tissues and in 786-O and ACHN cells compared with normal renal epithelial cells (HK-2). MIS18BP1 expression level was positively correlated with TNM stage and pathological grade, and showed good diagnostic performance for KIRC (AUC=0.693-0.885). WGCNA and enrichment analyses indicated that MIS18BP1-related genes were significantly enriched in the PI3K/AKT signaling pathway. In cultured renal cancer cells, MIS18BP1 knockdown significantly inhibited the proliferation, migration, and colony-forming ability of the cells, resulting also in decreased expression of N-cadherin and vimentin, increased expression of E-cadherin, and significantly lowered phosphorylation levels of PI3K and AKT. Treatment with 740Y-P partially reversed the inhibitory effects of MIS18BP1 knockdown on cell proliferation and migration. Conclusion MIS18BP1 is overexpressed in renal cancer and is associated with unfavorable clinical features. High expression of MIS18BP1 enhances proliferation and migration of renal cancer cells possibly by promoting EMT via activating the PI3K/AKT signaling pathway.

Key words: MIS18BP1, clear cell renal cell carcinoma, PI3K/AKT signaling pathway, epithelial-mesenchymal transition, cell proliferation, cell migration

Baofeng ZHAO, Xueying TAN, Xuewen LIANG, Yuyao PAN, Jiangshui YUAN, Weiqing SONG. MIS18BP1 regulates epithelial-mesenchymal transition, proliferation, and migration of renal cancer cells via the PI3K/AKT pathway[J]. Journal of Southern Medical University, 2026, 46(9): 2159-2172.

Figures/Tables 12

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.

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.

Tab.1 Diagnostic performance of MIS18BP1 for KIRC in TCGA and GEO datasets

Dataset	AUC	Best cutoff	Sensitivity(%)	Specificity(%)	Youden index
TCGA	0.693	2.097	60.07	76.39	0.3646
GSE36895	0.816	5.017	89.66	65.22	0.5488
GSE53757	0.879	6.759	84.72	77.78	0.6250
GSE66272	0.885	0.0806	80.77	92.59	0.7336

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.

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).

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.

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.

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.

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.

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.

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.

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).

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