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[Influence of crocin on gene expression profile of human bladder cancer cell lines T24]
Chun-Fang Lv1, Chun-Li Luo, Hui-Ying Ji
1Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, Chongqing Medical University, Chongqing 400016, China.
Objective:
To investigate the changes of gene expression profile in transitional cell carcinoma of bladder T24 cell after crocin treatment, in order to find the possible crocin targets.
Method:
The bladder cancer T24 cell line was treated with crocin. MTT assay was adopted to determine the inhibition rate for selecting the best effect time and concentration of crocin. Differentially expressed genes on groups with or without treatment of crocin were screened with high throughout cDNA microarray. One up-regulated gene p21(WAF1) and one down-regulated gene cyclinD1 were selected to undergo analysis by the reverse transcription polymerase chain reaction (RT-PCR). Moreover, immunocytochemical method was used to evaluate p21(WAF1) and cyclinD1 protein expression.
Result:
The growth of T24 cells was inhibited remarkably following a marked positive correlation between crocin concentration, time and inhibitor rate. When 3 mmol x L(-1) crocin treated T24 cells for 48h, the difference was significant compared with the control group (P < 0.05). Crocin induced wide changes of the gene expression profile of T24 cells. A total of 836 genes were up-regulated or down-regulated by more than 2 times, which were involved cell cycle controlling, DNA cell apoptosis, replication factor, and so on. The mRNA expression of p21(WAF1) and cyclinD1 detected by RT-PCR were in accordance with cDNA microarray data. The results of immunocytochemical method showed that p21(WAF1) and cyclinD1 protein expression were consistent with those mRNA expression.
Conclusion:
Crocin can induce the significant alteration of gene expression profile of T24 cell. It is suggested that the widly konwn anti-tumor effects of crocin are medicated at least in part by regulating the cell cycle controlling gene expression.
Insights
Crocin treatment significantly alters gene expression in bladder cancer T24 cells, impacting cell cycle control and apoptosis. This suggests crocin
Area of Science:
- Oncology
- Molecular Biology
- Pharmacology
Background:
- Transitional cell carcinoma of the bladder (TCCB) is a significant health concern.
- Crocin, a natural compound, exhibits potential anti-tumor properties.
- Understanding crocin's molecular mechanisms in TCCB is crucial for therapeutic development.
Purpose of the Study:
- To investigate the gene expression profile changes in T24 bladder cancer cells following crocin treatment.
- To identify potential molecular targets of crocin in TCCB.
Main Methods:
- T24 bladder cancer cells were treated with varying concentrations and durations of crocin.
- MTT assay was used to determine optimal treatment conditions.
- High-throughput cDNA microarray analysis identified differentially expressed genes.
- Reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemistry validated key gene and protein expression (p21(WAF1), cyclinD1).
Main Results:
- Crocin treatment significantly inhibited T24 cell growth in a dose- and time-dependent manner (P < 0.05 at 3 mmol/L for 48h).
- Crocin induced widespread changes in gene expression, with 836 genes showing over two-fold up- or down-regulation.
- Affected genes were involved in critical cellular processes including cell cycle control, apoptosis, and DNA replication.
- mRNA and protein expression of p21(WAF1) (up-regulated) and cyclinD1 (down-regulated) were consistent with microarray data.
Conclusions:
- Crocin significantly alters the gene expression profile of T24 bladder cancer cells.
- Crocin's anti-tumor effects are, at least partly, mediated by the regulation of cell cycle control gene expression.
- These findings highlight crocin as a potential therapeutic agent for bladder cancer.
