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The oncofetal gene survivin is re-expressed in osteoarthritis and is required for chondrocyte proliferation in vitro.

Lechler P, Balakrishnan S, Schaumburger J, Grässel S, Baier C, Grifka J, Straub RH, Renkawitz T - BMC Musculoskelet Disord (2011)

Bottom Line: Suppression of survivin resulted in inhibition of cell cycle progression and sensitization toward apoptotic stimuli in vitro.The present study indicates a role for survivin in osteoarthritic cartilage and human chondrocytes.Learning more about the functions of survivin in chondrocyte biology might further help toward understanding and modulating the complex processes of cartilage pathology and regeneration.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Orthopedic Surgery, University of Regensburg, Asklepios Klinikum Bad Abbach, Kaiser Karl V Allee 3, 93077 Bad Abbach, Germany. p.lechler@asklepios.com

ABSTRACT

Background: Regulation of cell death and cell division are key processes during chondrogenesis and in cartilage homeostasis and pathology. The oncogene survivin is considered to be critical for the coordination of mitosis and maintenance of cell viability during embryonic development and in cancer, and is not detectable in most adult differentiated tissues and cells. We analyzed survivin expression in osteoarthritic cartilage and its function in primary human chondrocytes in vitro.

Methods: Survivin expression was analyzed by immunoblotting and quantitative real-time PCR. The localization was visualized by immunofluorescence. Survivin functions in vitro were investigated by transfection of a specific siRNA.

Results: Survivin was expressed in human osteoarthritic cartilage, but was not detectable in macroscopically and microscopically unaffected cartilage of osteoarthritic knee joints. In primary human chondrocyte cultures, survivin was localized to heterogeneous subcellular compartments. Suppression of survivin resulted in inhibition of cell cycle progression and sensitization toward apoptotic stimuli in vitro.

Conclusions: The present study indicates a role for survivin in osteoarthritic cartilage and human chondrocytes. In vitro experiments indicated its involvement in cellular division and viability. Learning more about the functions of survivin in chondrocyte biology might further help toward understanding and modulating the complex processes of cartilage pathology and regeneration.

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Survivin expression in primary human chondrocytes. (A) Immunoblot for survivin protein in five independent primary chondrocyte cultures (Patient No.). Protein lysates at 24 hours after transfection of a GFP siRNA and at 24 and 48 hours after transfection of a survivin-specific siRNA are shown. (B) Relative expression of survivin RNA after transfection of the specific siRNA detected by quantitative PCR. One representative chondrocyte culture is shown.
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Figure 2: Survivin expression in primary human chondrocytes. (A) Immunoblot for survivin protein in five independent primary chondrocyte cultures (Patient No.). Protein lysates at 24 hours after transfection of a GFP siRNA and at 24 and 48 hours after transfection of a survivin-specific siRNA are shown. (B) Relative expression of survivin RNA after transfection of the specific siRNA detected by quantitative PCR. One representative chondrocyte culture is shown.

Mentions: Survivin expression in primary human chondrocyte cultures was analyzed by immunoblotting and quantitative real-time PCR. Survivin protein was expressed in all cultures established at passage 2 (n = 5) as detected by immunoblotting (Figure 2A). Antibody specificity was confirmed by transfection of a survivin-specific siRNA, which led to a significant reduction in the detectable survivin protein after 24 hours (Figure 2A). Equal loading was controlled by β-actin detection and Coomassie brilliant blue staining of the membranes (data not shown). Survivin expression at the protein level showed a marked decrease at 48 hours after the siRNA transfection. Knockdown of GFP did not result in alterations of the survivin protein levels. Next, we analyzed survivin mRNA expression by applying quantitative real-time PCR. Survivin was detectable in all cultures analyzed (n = 4) and knockdown of survivin resulted in a marked reduction in detectable survivin RNA (Figure 2B).


The oncofetal gene survivin is re-expressed in osteoarthritis and is required for chondrocyte proliferation in vitro.

Lechler P, Balakrishnan S, Schaumburger J, Grässel S, Baier C, Grifka J, Straub RH, Renkawitz T - BMC Musculoskelet Disord (2011)

Survivin expression in primary human chondrocytes. (A) Immunoblot for survivin protein in five independent primary chondrocyte cultures (Patient No.). Protein lysates at 24 hours after transfection of a GFP siRNA and at 24 and 48 hours after transfection of a survivin-specific siRNA are shown. (B) Relative expression of survivin RNA after transfection of the specific siRNA detected by quantitative PCR. One representative chondrocyte culture is shown.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3141611&req=5

Figure 2: Survivin expression in primary human chondrocytes. (A) Immunoblot for survivin protein in five independent primary chondrocyte cultures (Patient No.). Protein lysates at 24 hours after transfection of a GFP siRNA and at 24 and 48 hours after transfection of a survivin-specific siRNA are shown. (B) Relative expression of survivin RNA after transfection of the specific siRNA detected by quantitative PCR. One representative chondrocyte culture is shown.
Mentions: Survivin expression in primary human chondrocyte cultures was analyzed by immunoblotting and quantitative real-time PCR. Survivin protein was expressed in all cultures established at passage 2 (n = 5) as detected by immunoblotting (Figure 2A). Antibody specificity was confirmed by transfection of a survivin-specific siRNA, which led to a significant reduction in the detectable survivin protein after 24 hours (Figure 2A). Equal loading was controlled by β-actin detection and Coomassie brilliant blue staining of the membranes (data not shown). Survivin expression at the protein level showed a marked decrease at 48 hours after the siRNA transfection. Knockdown of GFP did not result in alterations of the survivin protein levels. Next, we analyzed survivin mRNA expression by applying quantitative real-time PCR. Survivin was detectable in all cultures analyzed (n = 4) and knockdown of survivin resulted in a marked reduction in detectable survivin RNA (Figure 2B).

Bottom Line: Suppression of survivin resulted in inhibition of cell cycle progression and sensitization toward apoptotic stimuli in vitro.The present study indicates a role for survivin in osteoarthritic cartilage and human chondrocytes.Learning more about the functions of survivin in chondrocyte biology might further help toward understanding and modulating the complex processes of cartilage pathology and regeneration.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Orthopedic Surgery, University of Regensburg, Asklepios Klinikum Bad Abbach, Kaiser Karl V Allee 3, 93077 Bad Abbach, Germany. p.lechler@asklepios.com

ABSTRACT

Background: Regulation of cell death and cell division are key processes during chondrogenesis and in cartilage homeostasis and pathology. The oncogene survivin is considered to be critical for the coordination of mitosis and maintenance of cell viability during embryonic development and in cancer, and is not detectable in most adult differentiated tissues and cells. We analyzed survivin expression in osteoarthritic cartilage and its function in primary human chondrocytes in vitro.

Methods: Survivin expression was analyzed by immunoblotting and quantitative real-time PCR. The localization was visualized by immunofluorescence. Survivin functions in vitro were investigated by transfection of a specific siRNA.

Results: Survivin was expressed in human osteoarthritic cartilage, but was not detectable in macroscopically and microscopically unaffected cartilage of osteoarthritic knee joints. In primary human chondrocyte cultures, survivin was localized to heterogeneous subcellular compartments. Suppression of survivin resulted in inhibition of cell cycle progression and sensitization toward apoptotic stimuli in vitro.

Conclusions: The present study indicates a role for survivin in osteoarthritic cartilage and human chondrocytes. In vitro experiments indicated its involvement in cellular division and viability. Learning more about the functions of survivin in chondrocyte biology might further help toward understanding and modulating the complex processes of cartilage pathology and regeneration.

Show MeSH
Related in: MedlinePlus