A cell-permeable dominant-negative survivin protein induces apoptosis and sensitizes prostate cancer cells to TNF-α therapy

Abstract

BackgroundSurvivin is a member of the inhibitor-of-apoptosis (IAP) family which is widely expressed by many different cancers. Overexpression of survivin is associated with drug resistance in cancer cells, and reduced patient survival after chemotherapy and radiotherapy. Agents that antagonize the function of survivin hold promise for treating many forms of cancer. The purpose of this study was to investigate whether a cell-permeable dominant-negative survivin protein would demonstrate bioactivity against prostate and cervical cancer cells grown in three dimensional culture.ResultsA dominant-negative survivin (C84A) protein fused to the cell penetrating peptide poly-arginine (R9) was expressed in E. coli and purified by affinity chromatography. Western blot analysis revealed that dNSurR9-C84A penetrated into 3D-cultured HeLa and DU145 cancer cells, and a cell viability assay revealed it induced cancer cell death. It increased the activities of caspase-9 and caspase-3, and rendered DU145 cells sensitive to TNF-α via by a mechanism involving activation of caspase-8.ConclusionsThe results demonstrate that antagonism of survivin function triggers the apoptosis of prostate and cervical cancer cells grown in 3D culture. It renders cancer cells sensitive to the proapoptotic affects of TNF-α, suggesting that survivin blocks the extrinsic pathway of apoptosis. Combination of the biologically active dNSurR9-C84A protein or other survivin antagonists with TNF-α therapy warrants consideration as an approach to cancer therapy.

Nanoformulated cell-penetrating survivin mutant and its dual actions

“…These results were consistent with our earlier results of the antitumor activities of SurR9-C84A. 12 Owing to the proliferative potential of SurR9-C84A in neurons with a low endogenous pool of survivin, differentiated SK-N-SH cells exhibited upregulation of cell-division markers. Endogenous survivin levels increased up to 46.3%, while PCNA and Ki67 showed a 5.1% and 24.9% increment, respectively.…”

“…18,43 Several reports have confirmed that agents acting against survivin, such as small-molecule inhibitors, antagonists, and antisense agents, interfere with survivin expression or function and suppress tumor progression. 12,18,44,45 Studies have also reported that survivin overexpression is responsible for the resistance developed in advanced tumors. 5,46 Therefore, in a clinical setting, interfering with the functions of wild-type survivin results in strong antitumor activity, and for this purpose baculovirus inhibitory apoptotic protein repeat (BIR)-motif mutant SurR9-C84A-loaded NPs were employed against the neuroblastoma cell lines.…”

Competitive inhibition of survivin using a cell-permeable recombinant protein induces cancer-specific apoptosis in colon cancer model

“…Abbreviations: sr9, cell-permeable dominant negative survivin surr9-c84a; chNP, chitosan nanoparticles; 3D, three-dimensional; smac, second mitochondria-derived activator of caspases; cIaP, cellular inhibitor of apoptosis. Although studies have been conducted with the C84A form of dominant negative survivin in both gene therapy 23 and protein therapy, 25 no interaction between the two forms of protein has been demonstrated. This is the first report that has showed interaction of dominant negative survivin protein SR9 with WT survivin in colon, liver, and breast cancer cells, and was further confirmed by Western blotting.…”

“…24 The apoptotic effects of SR9 have been evaluated in prostate cancer cells where it sensitized the cells to tumor necrosis factor alpha (TNF-α) and induced apoptosis. 25 Recently, we have shown that SR9 displays dual actions: it is cytotoxic to cancerous cells and helps in proliferation of normal non-cancerous cells. This is because tumor cells have a high endogenous survivin pool, and treatment with SR9 leads to forceful survivin expression.…”

Overexpressed genes in malignant pleural mesothelioma: implications in clinical management

“…Interestingly the vaccine strategy proposed in 2013 (55) effectively suppresses MPM tumor growth in vivo without induction of autoimmune response and the cytotoxic activity induced by this vaccine proved to be specific for MPM cells (57). Furthermore survivin expression proved to be linked to resistance to chemotherapeutic agents, including vincristine, cisplatin, bortezomib, tamoxifen, paclitaxel, TNF-a and TRAIL in tumour cells (52,54) and it is also responsible of the suppression of radiation-induced apoptosis (50). Thus survivin-targeting in combination with anti-cancer drug could be useful to enhance the effect of chemotherapeutic agents and may be promising for mesothelioma treatment.…”