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Effect of lentivirus-mediated survivin transfection on the morphology and apoptosis of nucleus pulposus cells derived from degenerative human disc in vitro.

Ma X, Lin Y, Yang K, Yue B, Xiang H, Chen B - Int. J. Mol. Med. (2015)

Bottom Line: The results revealed that the morphology of the NP cells derived from degenerative human disc transfected with LV‑mediated survivin was significantly altered as evidenced by cytomorphosis, the reduction of the cytoplasm and cell shrinkage.Furthermore, no significant anti-apoptotic effects were observed following LV‑mediated survivin transfection.However, cell morphology was evidently altered, whereas the apoptotic rate did not decrease.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedic Surgery, The Affiliated Hospital of Qingdao University Medical College, Qingdao University, Qingdao, Shandong 266003, P.R. China.

ABSTRACT
Lower back pain is a common concern, and 40% of all cases involve the degeneration of the intervertebral disc (IVD). However, the excessive apoptosis of disc cells plays an important role in IVD degeneration, particularly in the nucleus pulposus (NP). Thus, anti-apoptotic gene therapy to attenuate or reverse the degenerative process within the NP is being developed. Survivin is a unique inhibitor of apoptosis (IAP) and has been extensively investigated in cancer cells. However, little is known of the effects of survivin transfection on NP cells derived from degenerative human disc. In this study, we aimed to investigate the effects of lentivirus (LV)‑mediated survivin transfection on the morphology and apoptosis of NP cells derived from degenerative human disc in vitro. NP cells were transfected with LV‑mediated survivin. Subsequently, cell morphology was observed and the survivin mRNA expression levels were measured by RT‑qPCR. Apoptosis was analyzed by flow cytometry and by measuring caspase‑3 activity. The results revealed that the morphology of the NP cells derived from degenerative human disc transfected with LV‑mediated survivin was significantly altered as evidenced by cytomorphosis, the reduction of the cytoplasm and cell shrinkage. Following transfection, survivin gene expression significantly increased in the transfected cells and subsequent generation cells; however, no significant differences in the cell apoptotic rate and caspase‑3 activity were observed. We found that transfection of the survivin gene into NP cells led to the stable expression of survivin and induced marked changes in cell morphology. Furthermore, no significant anti-apoptotic effects were observed following LV‑mediated survivin transfection. Overall, our findings demonstrate that LV carrying surviving may be used to successfully enforce the expression of survivin in NP cells. However, cell morphology was evidently altered, whereas the apoptotic rate did not decrease. Comprehensive studies on the feasibility of using survivin in gene therapy in an aim to attenuate disc degeneration are warranted. Further research on the mechanisms responsible for the changes in cell morphology and cell function are also required.

No MeSH data available.


Related in: MedlinePlus

Schematic diagram of the mechanisms of survivin-mediated anti-apoptotic pathways.
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f1-ijmm-36-01-0186: Schematic diagram of the mechanisms of survivin-mediated anti-apoptotic pathways.

Mentions: Survivin is a unique inhibitor of apoptosis (IAP) that deters the activation of intrinsic and extrinsic pathways, with a focus on the former. Survivin inhibits apoptosis by binding to caspase-9 or by blocking second mitochondria-derived activator of caspases (SMAC; a pro-apoptotic protein that binds IAPs and thus prevents them from inhibiting caspases) and thus prevents the pro-apoptotic protein from blocking IAP proteins (Fig. 1) (23). The expression of survivin is strictly controlled in embryonic tissues and in the majority of tumors, but not during tissue differentiation and maturation (23,24). Thus, survivin presents an attractive target for cancer therapy (25), and has been extensively studied in cell cycle and apoptotic assays for cancer cells (26,27). Studies have demonstrated that the oncofetal gene, survivin, is re-expressed in osteoarthritis and rheumatoid arthritis (28–30). Moreover, preliminary studies have indicated that survivin is expressed in fetal disc tissue and have noted the differential expression of survivin between NP tissue derived from degenerative disc and that derived from a relatively normal disc (31,32). However, to our knowledge, limited research has been conducted on the effects of lentivirus (LV)-mediated survivin transfection on NP cells derived from degenerative human disc in vitro.


Effect of lentivirus-mediated survivin transfection on the morphology and apoptosis of nucleus pulposus cells derived from degenerative human disc in vitro.

Ma X, Lin Y, Yang K, Yue B, Xiang H, Chen B - Int. J. Mol. Med. (2015)

Schematic diagram of the mechanisms of survivin-mediated anti-apoptotic pathways.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4494593&req=5

f1-ijmm-36-01-0186: Schematic diagram of the mechanisms of survivin-mediated anti-apoptotic pathways.
Mentions: Survivin is a unique inhibitor of apoptosis (IAP) that deters the activation of intrinsic and extrinsic pathways, with a focus on the former. Survivin inhibits apoptosis by binding to caspase-9 or by blocking second mitochondria-derived activator of caspases (SMAC; a pro-apoptotic protein that binds IAPs and thus prevents them from inhibiting caspases) and thus prevents the pro-apoptotic protein from blocking IAP proteins (Fig. 1) (23). The expression of survivin is strictly controlled in embryonic tissues and in the majority of tumors, but not during tissue differentiation and maturation (23,24). Thus, survivin presents an attractive target for cancer therapy (25), and has been extensively studied in cell cycle and apoptotic assays for cancer cells (26,27). Studies have demonstrated that the oncofetal gene, survivin, is re-expressed in osteoarthritis and rheumatoid arthritis (28–30). Moreover, preliminary studies have indicated that survivin is expressed in fetal disc tissue and have noted the differential expression of survivin between NP tissue derived from degenerative disc and that derived from a relatively normal disc (31,32). However, to our knowledge, limited research has been conducted on the effects of lentivirus (LV)-mediated survivin transfection on NP cells derived from degenerative human disc in vitro.

Bottom Line: The results revealed that the morphology of the NP cells derived from degenerative human disc transfected with LV‑mediated survivin was significantly altered as evidenced by cytomorphosis, the reduction of the cytoplasm and cell shrinkage.Furthermore, no significant anti-apoptotic effects were observed following LV‑mediated survivin transfection.However, cell morphology was evidently altered, whereas the apoptotic rate did not decrease.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedic Surgery, The Affiliated Hospital of Qingdao University Medical College, Qingdao University, Qingdao, Shandong 266003, P.R. China.

ABSTRACT
Lower back pain is a common concern, and 40% of all cases involve the degeneration of the intervertebral disc (IVD). However, the excessive apoptosis of disc cells plays an important role in IVD degeneration, particularly in the nucleus pulposus (NP). Thus, anti-apoptotic gene therapy to attenuate or reverse the degenerative process within the NP is being developed. Survivin is a unique inhibitor of apoptosis (IAP) and has been extensively investigated in cancer cells. However, little is known of the effects of survivin transfection on NP cells derived from degenerative human disc. In this study, we aimed to investigate the effects of lentivirus (LV)‑mediated survivin transfection on the morphology and apoptosis of NP cells derived from degenerative human disc in vitro. NP cells were transfected with LV‑mediated survivin. Subsequently, cell morphology was observed and the survivin mRNA expression levels were measured by RT‑qPCR. Apoptosis was analyzed by flow cytometry and by measuring caspase‑3 activity. The results revealed that the morphology of the NP cells derived from degenerative human disc transfected with LV‑mediated survivin was significantly altered as evidenced by cytomorphosis, the reduction of the cytoplasm and cell shrinkage. Following transfection, survivin gene expression significantly increased in the transfected cells and subsequent generation cells; however, no significant differences in the cell apoptotic rate and caspase‑3 activity were observed. We found that transfection of the survivin gene into NP cells led to the stable expression of survivin and induced marked changes in cell morphology. Furthermore, no significant anti-apoptotic effects were observed following LV‑mediated survivin transfection. Overall, our findings demonstrate that LV carrying surviving may be used to successfully enforce the expression of survivin in NP cells. However, cell morphology was evidently altered, whereas the apoptotic rate did not decrease. Comprehensive studies on the feasibility of using survivin in gene therapy in an aim to attenuate disc degeneration are warranted. Further research on the mechanisms responsible for the changes in cell morphology and cell function are also required.

No MeSH data available.


Related in: MedlinePlus