Exon 2 methylation inhibits hepaCAM expression in transitional cell carcinoma of the bladder
Cuicui Pan1, Xiaohou Wu, Chunli Luo
1Department of Laboratory Medical Diagnostics, Ministry of Education, Chongqing Medical University, Chongqing, China.
Aim:
We aimed to investigate the mechanisms of hepaCAM inactivation in transitional cell carcinoma of the bladder through the analysis of hepaCAM exon 2 methylation.
Methods:
The methylation of hepaCAM exon 2 and the expression of hepaCAM were determined by methylation-specific restriction PCR assay and RT-PCR in bladder cancer cells (T24, BIU-87) as well as in 55 paired bladder cancer specimens. The methylated bladder cancer cells were treated with 5-Aza- 2'-deoxycytidine (5-Aza-CdR), a demethylating agent. MTT was used to detect the proliferation of T24 and BIU-87 cells.
Results:
The proliferation of T24 and BIU-87 cells was suppressed by treatment with different concentrations of 5-Aza-CdR; the expression of hepaCAM was absent in T24 and BIU-87 cells, and we found that exon 2 of hepaCAM was methylated in the 2 cells. hepaCAM mRNA was re-expressed and the methylation status of hepaCAM exon 2 was reversed after treatment with 5-Aza-CdR. The expression of hepaCAM mRNA in bladder cancer tissues was significantly lower than that in adjacent tissues. The methylation rate of hepaCAM exon 2 was significantly higher in bladder cancer tissues than in adjacent tissues. The methylation of hepaCAM exon 2 was related to hepaCAM expression in bladder cancer tissues.
Conclusions:
Downregulation of hepaCAM expression plays an important role in the tumorigenesis and development of bladder cancer. DNA methylation may be important for downregulation of hepaCAM expression in bladder cancer.
Insights
HepaCAM exon 2 methylation inactivates hepaCAM in bladder cancer, leading to tumor development. Demethylation therapy with 5-Aza-CdR reactivates hepaCAM, suppressing cancer cell proliferation.
Area of Science:
- Oncology
- Molecular Biology
- Epigenetics
Background:
- Transitional cell carcinoma of the bladder is a significant health concern.
- The role of hepaCAM in bladder cancer pathogenesis requires further elucidation.
- Epigenetic modifications, particularly DNA methylation, are implicated in cancer development.
Purpose of the Study:
- To investigate the mechanisms of hepaCAM inactivation in bladder transitional cell carcinoma.
- To analyze the methylation status of hepaCAM exon 2 and its correlation with hepaCAM expression.
- To assess the therapeutic potential of demethylating agents in restoring hepaCAM expression.
Main Methods:
- Analysis of hepaCAM exon 2 methylation using methylation-specific restriction PCR assay.
- Quantification of hepaCAM mRNA expression via RT-PCR in bladder cancer cell lines and patient specimens.
- Treatment of methylated bladder cancer cells with 5-Aza-2'-deoxycytidine (5-Aza-CdR) and assessment of cell proliferation using MTT assay.
Main Results:
- hepaCAM expression was absent in bladder cancer cell lines (T24, BIU-87) with methylated hepaCAM exon 2.
- Treatment with 5-Aza-CdR reversed hepaCAM exon 2 methylation and re-expressed hepaCAM mRNA, suppressing cell proliferation.
- Significantly lower hepaCAM mRNA expression and higher hepaCAM exon 2 methylation rates were observed in bladder cancer tissues compared to adjacent tissues.
Conclusions:
- Downregulation of hepaCAM expression is crucial in the tumorigenesis and progression of bladder cancer.
- DNA methylation of hepaCAM exon 2 is a key mechanism contributing to hepaCAM downregulation in bladder cancer.
- Targeting DNA methylation may represent a viable therapeutic strategy for bladder cancer.
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