pubmed.ncbi.nlm.nih.gov

Targeting of the anti-apoptotic gene survivin in human thyroid carcinoma - PubMed

Targeting of the anti-apoptotic gene survivin in human thyroid carcinoma

Zhouxun Chen et al. Int J Mol Med. 2012 Sep.

Abstract

Survivin is a novel apoptosis inhibitor. Its gene is related to the baculovirus gene, which is believed to play a crucial role in fetal development and in cancer. We attempted to determine the expression of survivin in both thyroid goiter and carcinoma tissues, and to evaluate its prognostic value in human thyroid disease. In the present study, we applied small interfering RNA (siRNA) directed against survivin to determine the effects of decreasing the high constitutive levels of this protein in the FTC-133 thyroid follicular cancer cell line. Using reverse transcription PCR and immunohistochemistry, we compared the expression of survivin with relevant clinical and pathological data of 90 postsurgical specimens from patients with primary thyroid carcinoma and patients with benign goiter (33 with papillary thyroid cancer, 24 with follicular thyroid cancer, 18 with undifferentiated thyroid cancer and 15 cases with goiter). For the siRNA treatment in a human follicular thyroid carcinoma cell line, fluorescein-labeled double-stranded ultrapure siRNAs were used. RT-PCR identified the survivin transcript in 67/75 (89.3%) tumor samples and in 4/15 benign goiter samples. Immunohistochemical analysis showed positive immunoreactivity in 65/75 (86.7%) carcinomas while no expression was noted in all of the 15 benign goiter tissues. Survivin mRNA and protein levels were significantly higher in cancer tissues compared to benign goiter tissues (P<0.001). Higher survivin expression was found in the tumor tissues of pT3/pT4 and in the tumors with lymph node metastasis (P<0.05). Tumors with distant metastasis demonstrated higher survivin expression compared to the tumors without distant metastasis. Additionally, the expression of survivin in undifferentiated carcinomas was higher than that in differentiated ones. There was no significant correlation between survivin expression and age, gender, histological subtype and pathological stage. Our additional studies demonstrated that siRNA directed against survivin markedly decreased the protein expression of survivin. In conclusion, we conclude that survivin expression indicates more aggressive behavior and metastatic ability in thyroid cancer cells in vivo. Survivin can be used as a diagnostic and therapeutic marker for thyroid carcinoma and an important target in the strategy of thyroid cancer therapy. Our results of siRNA silencing indicate that siRNA may have potential as a therapeutic modality in the treatment of human thyroid cancer.

PubMed Disclaimer

Figures

Figure 1.
Figure 1.

Survivin mRNA expression in thyroid tissues. (a) Weak or negative expression of survivin mRNA in benign goiter tissues; (b) strong survivin mRNA expression in carcinoma tissues. K+, positive control, human colorectal carcinoma cell line ‘Caco’.

Figure 2.
Figure 2.

Correlation between the transcript expression of endogenous survivin and various clinicopathological parameters. Thyroid cancer samples expressed higher transcript expression of endogenous survivin when compared to the benign goiters; the expression was significantly correlated with the grade of tumor invasiveness (pT), local lymph node metastasis (N) and distant metastasis (M) of the thyroid carcinoma tissues.

Figure 3.
Figure 3.

Immunostaining patterns of survivin in frozen human thyroid tissue samples. (a) Thyroid goiter tissue with a loss of expression of survivin. (b) Strong immunostaining in a primary PTC (pT4N0M0). (c) A primary FTC (pT4N1M0) showing moderate survivin protein expression. (d) Uniformly intense staining of survivin protein expression in UTC (pT4N1M0) tissue. PTC, papillary thyroid cancer; FTC, follicular thyroid cancer; UTC, undifferentiated thyroid cancer.

Figure 4.
Figure 4.

Correlation between the protein expression of endogenous survivin and various clinicopathological parameters. Protein expression of endogenous survivin was significantly correlated with the grade of tumor invasiveness T and local lymph node metastasis (N); and with distant metastasis (M) of thyroid carcinoma tissues but this value did not achieve statistical significance.

Figure 5.
Figure 5.

Transfection efficiency of siRNA of FTC-133 cells. Approximately 60% of whole cells were transfected with fluorescein-labeled siRNA (upper left panel, x100 magnification; upper right panel, x200 magnification). A ginkgo biloba formed single transfected cell and a spindle-shaped cell are shown in the lower panels (x400 magnification).

Figure 6.
Figure 6.

Effect of survivin siRNA on FTC-133 cells as detected using immunocytochemistry. (a) Untreated cells; (b) cells treated with mismatch siRNA; (c) cells treated with sis-100 siRNA; (d) cells treated with sis-248 siRNA

Figure 7.
Figure 7.

Western blot analysis indicates reduced survivin expression following siRNA treatment in FTC-133 cells.

Similar articles

Cited by

References

    1. Wyllie AH. Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature. 1980;284:555–556. - PubMed
    1. Nagata S. Apoptosis by death factor. Cell. 1997;88:355–365. - PubMed
    1. Thompson CB. Apoptosis in the pathogenesis and treatment of disease. Science. 1995;267:1456–1462. - PubMed
    1. Sachs L, Loterm J. Control of programmed cell death in normal and leukemic cells: new implications for therapy. Blood. 1993;82:15–21. - PubMed
    1. Yang E, Korsmeyer SJ. Molecular thanatopsis: a discourse on the bcl-2 family and cell death. Blood. 1996;88:1456–1462. - PubMed

Publication types

MeSH terms

Substances