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. 2015 Jul;10(1):330-334.
doi: 10.3892/etm.2015.2466. Epub 2015 Apr 30.

Expression of survivin in adenoid cystic carcinoma of the lacrimal gland and the effect of intervention with arsenic trioxide in vitro

Yingzhe Pan  1 , Yiqiao Xing  2 , Hui Wang  1
Affiliations

Expression of survivin in adenoid cystic carcinoma of the lacrimal gland and the effect of intervention with arsenic trioxide in vitro

Yingzhe Pan et al. Exp Ther Med. 2015 Jul.

Abstract

The aim of the present study was to investigate the role of survivin in adenoid cystic carcinoma of the lacrimal gland (LGACC) and the effect of arsenic trioxide (As2O3) intervention in vitro. In total, 13 patients with LGACC were recruited from Renmin Hospital of Wuhan University between 2007 and 2012. The patients were divided into different groups, namely tumor-node-metastasis (TNM)1-2 and TNM3-4, according to the TNM classification. A total of eight samples were included in the TNM1-2 group, while five samples were included in the TNM3-4 group. In addition, six patients that suffered from other diseases (no tumor), but underwent eye surgery, were selected as the controls. The mRNA and protein expression levels of survivin were analyzed. In addition, primary adenoid cystic carcinoma (ACC) cells were cultured and the expression of survivin was silenced using short interfering RNA (siRNA), after which the cell proliferation was determined. Furthermore, the primary cultured ACC cells were treated with different doses of As2O3 to observe the effect on the apoptosis rate. The mRNA and protein expression levels of survivin in the LGACC groups were higher when compared with the controls (P<0.01). In addition, the expression levels were higher in the TNM3-4 group when compared with the TNM1-2 group (P<0.01). After silencing survivin expression using siRNA, the rate of ACC cell proliferation was shown to decrease at days 2, 3, 4 and 5 following culture, particularly in the TNM3-4 group at days 2 and 3 (P<0.05), day 4 (P<0.01 vs. TNM1-2 + siRNA); and days 3 and 5 (P<0.05), and day 2 (P<0.01 vs. TNM3-4 + siRNA). The mRNA expression levels of survivin in the TNM1-2 and TNM3-4 groups were significantly decreased following treatment with the various doses of As2O3 (2, 4 and 6 µΜ) for 48 h, and the apoptosis rate of the ACC cells was markedly increased (TNM1-2 and 3-4: As2O3 6 µM vs. As2O3 2 µM, As2O3 6 µM vs. As2O3 4 Μm, P<0.01; TNF3-4, As2O3 4 µM VS. As2O3 2 µM, P<0.01; TNM1-2, As2O3 4 µM vs. As2O3 2 µM, P<0.05). Therefore, survivin was demonstrated to be highly expressed in ACC cells of the lacrimal gland, and inhibition of survivin gene expression was found to suppress the proliferation of the ACC cells. Furthermore, As2O3 treatment was demonstrated to inhibit the mRNA expression of survivin in the ACC cells, while significantly increasing the apoptosis rate.

Keywords: adenoid cystic carcinoma; arsenic trioxide; lacrimal gland; survivin.

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Figures

Figure 1.
Figure 1.
Relative mRNA and protein expression levels of survivin in human adenoid cystic carcinoma tissues from the lacrimal gland. Results are expressed as the mean ± standard deviation (n=5). **P<0.01, vs. control; ##P<0.01, vs. TNM1–2. TNM, tumor-node-metastasis.
Figure 2.
Figure 2.
Relative mRNA expression levels of survivin in primary human adenoid cystic carcinoma cells from the lacrimal gland following treatment with siRNA for 48 h. Results are expressed as the mean ± standard deviation (n=5). **P<0.01, vs. TNM1–2; ##P<0.01, vs. TNM3–4. TNM, tumor-node-metastasis; siRNA, short interfering RNA.
Figure 3.
Figure 3.
Integrated optical density values obtained in the MTT assay prior to and following siRNA treatment in the TNM1–2 and TNM3–4 cells. Results are expressed as the mean ± standard deviation (n=5). *P<0.05 and **P<0.01, vs. TNM(–2) + siRNA; #P<0.05 and ##P<0.01, vs. TNM(–4) + siRNA. TNM, tumor-node-metastasis; siRNA, short interfering RNA.
Figure 4.
Figure 4.
Relative mRNA expression levels of survivin in primary human adenoid cystic carcinoma cells from the lacrimal gland after 48 h treatment with As2O3. Results are expressed as the mean ± standard deviation (n=5). *P<0.05 and **P<0.01, vs. control; #P<0.05 and ##P<0.01, vs. As2O3 (2 µΜ) or As2O3 (4 µΜ). TNM, tumor-node-metastasis; As2O3, arsenic trioxide.
Figure 5.
Figure 5.
Treatment of primary human adenoid cystic carcinoma cells from the lacrimal gland with As2O3 resulted in a time- and dose-dependent increase in apoptotic cells. Results are expressed as the mean ± standard deviation (n=5). **P<0.01, As2O3 (6 µΜ) vs. As2O3 (2 µΜ); ##P<0.01, As2O3 (6 µΜ) vs. As2O3 (4 µΜ); &P<0.05 and &&P<0.01, As2O3 (4 µΜ) vs. As2O3 (2 µΜ). TNM, tumor-node-metastasis; As2O3, arsenic trioxide.
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