Expression of MCM4 in bladder cancer and its correlation with PCNA
10.3760/cma.j.cn121382-20240731-00611
- VernacularTitle:MCM4在膀胱癌中表达及与PCNA的相关性研究
- Author:
Yuhua WANG
1
;
Feng ZHANG
;
Yalin WANG
;
Jun LI
Author Information
1. 新乡医学院第三附属医院泌尿外科，新乡 453000
- Keywords:
Minichromosome maintenance protein 4;
Proliferating cell nuclear antigen;
Bladder cancer;
Cell proliferation
- From:
International Journal of Biomedical Engineering
2024;47(6):591-599
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the expression of minichromosome maintenance protein 4 (MCM4) in bladder cancer and its correlation with proliferating cell nuclear antigen (PCNA).Methods:The expression of MCM4 in bladder cancer tissues and adjacent normal tissues was analyzed in the gene expression profiling interactive analysis (GEPIA) database. Paraffin sections of bladder cancer tissues and adjacent normal tissues from 72 patients admitted to the Department of Urology, the Third Affiliated Hospital of Xinxiang Medical College from June 2020 to May 2022 were collected. The expression of MCM4 in bladder cancer tissues and adjacent normal tissues was examined by immunohistochemical staining. The relationship between MCM4 expression and clinicopathologic features and prognosis of bladder cancer patients was analyzed. Bladder cancer T24 and 5637 cells were cultured in vitro, and the control and transfection groups were established. The cells in the transfection group were transfected with short hairpin RNA (shRNA) to knock down MCM4. The expression of MCM4 was evaluated by real-time reverse transcription-PCR (RT-qPCR) and Western blotting. The effect of MCM4 on bladder cancer cell proliferation was evaluated by clonogenic assay, cell counting kit-8 (CCK-8) assay, and cell cycle assay. A nude mouse model was established using BALB/c nude mice and T24 cell, and the effect of MCM4 on tumor growth was evaluated by Western blotting. The correlation between MCM4 and PCNA was investigated by GEPIA database and immunohistochemical staining. Results:The GEPIA database analysis showed that the relative expression of MCM4 in bladder cancer tissues ( n=404) was higher than that in adjacent normal tissues ( n=28, P<0.05). The expression of MCM4 in bladder cancer tissues was higher than that in adjacent normal tissues, and no higher expression was observed in adjacent normal tissues. The expression level of MCM4 was closely correlated with tumor size in the low-expression group and high-expression group of patients ( χ2=10.892, P=0.001), but not with age ( χ2=1.583, P=0.208), gender ( χ2=0.011, P=0.915), and tumor differentiation ( χ2=0.196, P=0.658). The 5-year overall survival rate was significantly reduced in patients with high MCM4 expression ( P=0.013). Compared with the control group [(1.0±0.2), (1.0±0.2)], the expression of MCM4 gene [(0.4±0.1), (0.4±0.1)] in the transfection group of T24 and 5637 cells were lower (both P<0.05). Compared with the control group [(1.0±0.2), (1.0±0.2)], the expressions of MCM4 protein [(0.5±0.1), (0.3±0.1)] in the transfection group of T24 and 5637 cells were lower (both P<0.05). Compared with the control group [(150±16), (160±18) unit], the clonal cell number [(110±11), (120±12) unit] of T24 and 5637 cells in the transfection group were decreased (both P<0.05). Compared with the control group [(1.0±0.2), (1.0±0.2)], the absorbance ( A) value [(0.4±0.1), (0.5±0.1)] of T24 and 5637 cells in the transfection group were decreased (both P<0.05). Compared with the control group [(77±7)%, (67±7)%], the transfection group blocked the G 1/S phase [(89±8)%, (76±8)%] of T24 and 5637 cells (both P<0.05). Compared with the control group, the tumor volume in the transfection group was significantly smaller [(196±16) mm 3vs (304±25) mm 3, P<0.05], and the expression of MCM4 protein in T24 cell in the transfection group was lower [(0.5±0.1) vs (1.0±0.2), P<0.05]. GEPIA database analysis showed a correlation between MCM4 and PCNA ( R=0.61, P<0.05), and bladder cancer patients with high MCM4 expression also had similarly high PCNA expression. Conclusions:MCM4 is highly expressed in bladder cancer tissues and may promote colorectal cancer proliferation by affecting PCNA.
