Pleiotropic cell-division defects and apoptosis induced by interference with survivin function

Abstract

Here we investigate the role of the control of apoptosis in normal cell division. We show that interference with the expression or function of the apoptosis inhibitor survivin causes caspase-dependent cell death in the G2/M phase of the cell cycle, and a cell-division defect characterized by centrosome dysregulation, multipolar mitotic spindles and multinucleated, polyploid cells. Use of a dominant-negative survivin mutant or antisense survivin complementary DNA disrupts a supramolecular assembly of survivin, caspase-3 and the cyclin-dependent-kinase inhibitor p21Waf1/Cip1 within centrosomes, and results in caspase-dependent cleavage of p21. Polyploidy induced by survivin antagonists is accentuated in p21-deficient cells, and corrected by exogenous expression of p21. These findings show that control of apoptosis and preservation of p21 integrity within centrosomes by survivin are required for normal mitotic progression.

Knockdown of survivin expression by small interfering RNA reduces the clonogenic survival of human sarcoma cell lines independently of p53

“…Furthermore, clonogenic survival of cells from each line was reduced by 65-86% (Fig 1c, Table 1). By using antisense oligonucleotides, 8,[10][11][12][34][35][36] ribozymes, 9,13,37 and siRNA (in a colon cancer cell line) 23 to knock down expression of survivin mRNA and protein, other investigators have observed a remarkable reduction in survivin mRNA that ranged from 70 to 90% and a reduction in survivin protein that ranged from 60 to 90%. The study conducted by Williams et al 23 has described the application of survivin-specific siRNA, which differs from the construct applied in our study, resulting in a reduction of survivin protein in colon cancer.…”

“…This finding is in accordance with the results of other knockdown experiments. 8,10,12,23,25,26 However, the fate of these polyploid cells is not completely known. Chen and co-workers suggested that many of the polyploid cells undergo apoptosis; however, we and others 12,13 did not observe a large number of these cells undergoing apoptosis.…”

“…Treatment with either survivin-specific antisense oligonucleotides or ribozymes has resulted in reduced survivin expression, which has been correlated with a decrease in tumor growth in mice, with an increase in caspasedependent cell death (apoptosis), and with the formation of polyploid cells in vitro. [8][9][10][11][12][13][14][15][16] In addition to the use of antisense oligonucleotides and ribozymes, the application of small interfering RNA (siRNA) is an effective way to silence gene transcription. 17,18 Transcription silencing is thought to be a defense strategy that preserves genomes through evolution from attack by double-stranded RNA viruses and transposons.…”

Survivin expression in oral squamous cell carcinoma

“…Interference with survivin function induces pleiotropic celldivision defects and apoptosis (Li et al, 1999), suggesting a potential role at the interface between cell division and apoptosis control. In retrospective trials, survivin expression correlated with unfavourable neuroblastoma (Adida et al, 1998a), reduced overall survival in primitive colorectal (Kawasaki et al, 1998) and recurrent colorectal cancer (Sarela et al, 2000), non-small-cell lung cancer (Monzo et al, 1999), breast cancer (Tanaka et al, 2000), and increased rates of recurrences in bladder cancer (Swana et al, 1999).…”

Induction of apoptosis of non-small cell lung cancer by a methylated oligonucleotide targeting survivin gene

“…As NSCLC cells express the highest level of survivin found in humor tumor, 4 inhibition of survivin would block the anti-apoptosis and mitotic progression in NSCLC. The molecular antagonists of survivin, including antisense, ribozymes, siRNA or dominant negative mutants have been successfully utilized to inhibit survivin expression or to interfere with its functions, inducing apoptosis and inhibiting tumor growth in vitro and in vivo, [21][22][23][24][25][26][27][28][29] As there are only two survivin alleles in each cell compared with hundreds of thousands of survivin mRNA molecules, an antigene strategy at the transcriptional level probably possesses advantage over antisense, RNAi or ribozyme-based techniques. Recently, Yao et al 20 have devised a method to inhibit transcriptional initiation by constructing short methylated oligonucleotides which induce DNA methylation at specific loci.…”