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[Induced RAS association domain family gene 1A gene expression by arsenic trioxide in nasopharyngeal carcinoma cell]
Jun-Li Hu1, Yue-Fei Zhang, Zhi-Dong Qiu
1Department of Otorhinolaryngology Head and Neck Surgery, Affiliated Hospital of GuangDong Medical College, Zhanjiang 524023, China.
Objective:
To investigate the effect of arsenic trioxide (As2O3) on expression of anti-oncogene RAS association domain family gene 1A(RASSF1A) in nasopharyngeal carcinoma cell line CNE-2Z.
Methods:
CNE-2Z cells were treated with various concentrations of As2O3 for different times. The IC(50) values were detected by trypan blue stain assay. Cell cycle redistribution was analyzed by flow cytometry. The final concentration 2 micromol/L, 1 micromol/L, 0.5 micromol/L of As2O3 was added to CNE-2Z cell for succedent experiments. The controls and no drugs of CNE-2Z cells were cultivated for 48 h. Methylation specific PCR was used to detect the change of methylation status of RASSF1A gene. The expression of RASSF1A gene was detected by reverse transcription PCR and Western blot at mRNA and protein level.
Results:
The suppression of cell proliferation by As2O3 was time and dose-dependent. After being treated with As2O3, the IC(50) values of As2O3 were (1.50 +/- 0.05), (1.09 +/- 0.13), (0.65 +/- 0.04) micromol/L at 24, 48, and 72 h, respectively. As2O3 also arrested CNE-2Z cells in G2/M phase of cell cycle. After the effect of As2O3, the methylation of RASSF1A gene became weaker by increasing the concentration of As2O3; and the expression of RASSF1A gene became stronger at mRNA and protein level. Between different concentration of As2O3 group and no drugs group, the differences had statistical significance (P < 0.05). Along with increasing the concentration of As2O3, the expression of RASSF1A gene became stronger at mRNA and protein level, the methylation of RASSF1A gene became weaker and weaker.
Conclusions:
As2O3 can activate the expression of RASSF1A gene to inhibit the cell cycle progress of nasopharyngeal carcinoma cell line.
Insights
Arsenic trioxide (As2O3) was found to suppress nasopharyngeal carcinoma cell proliferation. As2O3 also activates the anti-oncogene RASSF1A, inhibiting cell cycle progression.
Area of Science:
- Oncology
- Molecular Biology
- Pharmacology
Background:
- Nasopharyngeal carcinoma (NPC) is a significant health concern.
- The anti-oncogene RAS association domain family gene 1A (RASSF1A) plays a crucial role in tumor suppression.
- Understanding the regulation of RASSF1A is vital for developing novel cancer therapies.
Purpose of the Study:
- To investigate the effect of arsenic trioxide (As2O3) on the expression of the RASSF1A gene in the CNE-2Z nasopharyngeal carcinoma cell line.
- To determine if As2O3 influences RASSF1A methylation status and gene expression at both mRNA and protein levels.
Main Methods:
- CNE-2Z cells were treated with varying concentrations of As2O3.
- Cell proliferation was assessed using trypan blue staining to determine IC50 values.
- Cell cycle analysis was performed via flow cytometry.
- RASSF1A gene methylation was analyzed using methylation-specific PCR.
- RASSF1A mRNA and protein expression were quantified using reverse transcription PCR and Western blot, respectively.
Main Results:
- As2O3 demonstrated a time- and dose-dependent suppression of CNE-2Z cell proliferation.
- As2O3 treatment led to cell cycle arrest at the G2/M phase.
- Increasing concentrations of As2O3 reduced RASSF1A gene methylation.
- Concurrently, As2O3 treatment significantly upregulated RASSF1A expression at both mRNA and protein levels (P < 0.05).
Conclusions:
- Arsenic trioxide effectively inhibits nasopharyngeal carcinoma cell proliferation.
- As2O3 activates RASSF1A gene expression, likely through demethylation.
- This activation of RASSF1A contributes to the inhibition of cell cycle progression in nasopharyngeal carcinoma cells.
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