Cancer Res Treat.  2004 Aug;36(4):255-262.

Comparison of As2O3 and As4O6 in the Detection of SiHa Cervical Cancer Cell Growth Inhibition Pathway

• Kim YW
• Bae SM
• Lee KH
• Lee JM
• Namkoong S
• Lee IP
• Kim CK
• Seo JS
• Sin JI
• Kim YW
• Ahn WS

Abstract

PURPOSE
An arsenical compound, As2O3, has been reported to be effective for treating acute leukemia and inducing apoptosis in many different tumor cells. In this study, the ability of As4O6 to suppress cell growth and induce gene expression patterns was tested using a cDNA microarray in HPV16 immortalized cervical carcinoma cells, SiHa cells, along with As2O3. MATERIALS AND METHODS: A novel arsenical compound, As4O6, was designed and its ability to induce cell growth inhibition as well as gene expression profiles along with As2O3 in HPV16 infected SiHa cervical cancer cells was compared. Both As2O3 and As4O6 induced apoptosis in SiHa cells, as determined by DNA ladder formation. To further compare the gene expression profiles between these two drugs, a 384 cDNA microarray system was employed. Also, the gene expression profiles were classified into the Gene Ontology (GO) to investigate apoptosis-related cellular processes. RESULTS: As4O6 was more effective i suppressing the growth of SiHa cells in vitro compared to As2O3. In the case of treatment with As2O3, 41 genes were up- or down- regulated at least 2 fold compared to non-treatment. However, 65 genes were up- or down-regulated by As4O6 treatment. In particular, 27 genes were commonly regulated by both arsenic compounds. Also, the GO analysis indicated that down-regulation of cell-regulatory functions, such as cell cycle, protein kinase activity and DNA repair, induced anti-tumor effect. CONCLUSION: These data support that As4O6 could be more effective than As2O3 in inhibiting the growth of HPV16 infected cervical cancer cells. This appears to be mediated through a unique, but overlapping regulatory mechanism(s), suggesting that the regulated genes and cellular processes could be further used as a new potential drug approach for treating cervical cancer in clinical settings.

Keyword

Cervical cancer; Arsenic compound; Apoptosis; cDNA microarray; Gene ontology

MeSH Terms

Apoptosis
Arsenicals
Cell Cycle
DNA
DNA Repair
Down-Regulation
Gene Expression
Gene Ontology
Leukemia
Oligonucleotide Array Sequence Analysis
Protein Kinases
Transcriptome
Uterine Cervical Neoplasms*
Arsenicals
DNA
Protein Kinases

Figure

• 

  Fig. 1 Induction of DNA ladder by As2O3 and As4O6 in SiHa cells. Cells were treated with the indicated amounts (0, 0.5, 1 and 2 µM) of As2O3 and As4O6 for 48 hrs. The DNA was analyzed on a 2% agarose gel and photographed under UV light.

• 

  Fig. 2 Effects of As2O3 and As4O6 on the growth suppression of SiHA cells in vitro. Cells were treated with the indicated amount of the arsenic compounds, As2O3 and As4O6, and incubated for 4 days. Cell growth suppression was measured as described in 'Methods and Materials'. The OD was measured at 405 nm. The assay was performed in triplicate and average OD values and SD were recorded. This was repeated two more times with similar results. *Statistically significant at p<0.05, using the paired Student's t test, compared to no drug treatment (control, CTL).

• 

  Fig. 3 Confirmations of cDNA chip experiment by Northern blot analysis. Total RNA obtained from As2O3 (lane 2) and As4O6 (lane 3) treated cells subjected to Northern analysis, as described in 'Materials and methods'.

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