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Efficient inhibition of human glioma development by RNA interference-mediated silencing of PAK5.

Gu X, Wang C, Wang X, Ma G, Li Y, Cui L, Chen Y, Zhao B, Li K - Int. J. Biol. Sci. (2015)

Bottom Line: In addition, silencing PAK5 expression in U87 cells weakened their colony formation ability and in vivo tumorigenesis ability.Further studies demonstrated that PAK5 inhibition led to an increase in cleaved caspase 3 and a decrease in β-catenin.In conclusion, our results suggest that the inhibition of PAK5 by RNA interference might efficiently suppress tumor development of glioma cells with PAK5 high expression.

View Article: PubMed Central - PubMed

Affiliation: 1. Institute of Neurology, Guangdong Medical College, Zhanjiang, China. ; 2. Guangdong Key Laboratory of Age-Related Cardiac and Cerebral Diseases, Affiliated Hospital of Guangdong Medical College, Zhanjiang, China.

ABSTRACT
Glioma is the most common type of primary intracranial tumor and is highly lethal due to its pathogenetic location, high invasiveness, and poor prognosis. Even combined surgery and chemoradiotherapy do not effectively rescue glioma patients. Molecular target therapy is considered a safe and promising therapy for glioma. The identification of a novel, effective target protein in gliomas is of great interest. We found that PAK5 was highly expressed in the tumor tissues of glioma patients and human glioma cell lines. We then used a lentivirus-delivered short hairpin RNA to stably silence PAK5 expression in glioma cells and explore its influence. The results showed that the inhibition of PAK5 reduced cell viability and delayed the cell cycle at the G0/G1 phase in the glioma cells with PAK5 high expression. In addition, silencing PAK5 expression in U87 cells weakened their colony formation ability and in vivo tumorigenesis ability. Further studies demonstrated that PAK5 inhibition led to an increase in cleaved caspase 3 and a decrease in β-catenin. In conclusion, our results suggest that the inhibition of PAK5 by RNA interference might efficiently suppress tumor development of glioma cells with PAK5 high expression. This finding provides a novel, promising therapeutic target for glioma treatment.

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Inhibition of PAK5 remarkably suppresses cell viability in glioma cells with high PAK5 expression. (A-C) Cell viability curves of U87 (A), SHG-44 (B) and CHG-5 (C) cells during the 5 days were evaluated by the MTT assay. The data represent the mean ± SEM of three independent experiments. *, P < 0.05 vs. cells infected with Scr-shRNA lentivirus. (D) PAK5 protein expression in U87 cells infected with a vector with expressing PAK5 after PAK5-shRNA1-mediated PAK5 knockdown. (E) Cell viability curves of U87 cells of three groups during the 5 days were evaluated by the MTT assay. The data represent the mean ± SEM of three independent experiments. *, P < 0.05 vs. cells infected with Scr-shRNA lentivirus.
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Figure 3: Inhibition of PAK5 remarkably suppresses cell viability in glioma cells with high PAK5 expression. (A-C) Cell viability curves of U87 (A), SHG-44 (B) and CHG-5 (C) cells during the 5 days were evaluated by the MTT assay. The data represent the mean ± SEM of three independent experiments. *, P < 0.05 vs. cells infected with Scr-shRNA lentivirus. (D) PAK5 protein expression in U87 cells infected with a vector with expressing PAK5 after PAK5-shRNA1-mediated PAK5 knockdown. (E) Cell viability curves of U87 cells of three groups during the 5 days were evaluated by the MTT assay. The data represent the mean ± SEM of three independent experiments. *, P < 0.05 vs. cells infected with Scr-shRNA lentivirus.

Mentions: We next explored the effects of PAK5 inhibition in glioma cells. First, we used a MTT assay to assess cell growth dynamics in Scr-shRNA- and PAK5-shRNA-infected cells. After 5 days, in U87 and SHG-44 cells, PAK5-shRNA-infected cells displayed remarkable cell growth inhibition compared with Scr-shRNA-infected cells. At day 5, the cell viability of the Scr-shRNA-infected group was 4 times higher than that of the PAK5-shRNA-infected groups (Fig. 3A and B). However, there was no significant difference between Scr-shRNA- and PAK5-shRNA-infected CHG-5 cells, cells with low PAK5 expression (Fig. 3C). To further determine the effect of PAK5 in glioma cells with high PAK5 expression, we enhanced PAK5 expression by transfecting PAK5 vector in PAK5-shRNA1-infected U87 cells (Fig. 3D). The results showed that enhancing PAK5 expression rescued the decrease of cell viability resulting from RNAi-mediated PAK5 knockdown (Fig. 3E).


Efficient inhibition of human glioma development by RNA interference-mediated silencing of PAK5.

Gu X, Wang C, Wang X, Ma G, Li Y, Cui L, Chen Y, Zhao B, Li K - Int. J. Biol. Sci. (2015)

Inhibition of PAK5 remarkably suppresses cell viability in glioma cells with high PAK5 expression. (A-C) Cell viability curves of U87 (A), SHG-44 (B) and CHG-5 (C) cells during the 5 days were evaluated by the MTT assay. The data represent the mean ± SEM of three independent experiments. *, P < 0.05 vs. cells infected with Scr-shRNA lentivirus. (D) PAK5 protein expression in U87 cells infected with a vector with expressing PAK5 after PAK5-shRNA1-mediated PAK5 knockdown. (E) Cell viability curves of U87 cells of three groups during the 5 days were evaluated by the MTT assay. The data represent the mean ± SEM of three independent experiments. *, P < 0.05 vs. cells infected with Scr-shRNA lentivirus.
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Figure 3: Inhibition of PAK5 remarkably suppresses cell viability in glioma cells with high PAK5 expression. (A-C) Cell viability curves of U87 (A), SHG-44 (B) and CHG-5 (C) cells during the 5 days were evaluated by the MTT assay. The data represent the mean ± SEM of three independent experiments. *, P < 0.05 vs. cells infected with Scr-shRNA lentivirus. (D) PAK5 protein expression in U87 cells infected with a vector with expressing PAK5 after PAK5-shRNA1-mediated PAK5 knockdown. (E) Cell viability curves of U87 cells of three groups during the 5 days were evaluated by the MTT assay. The data represent the mean ± SEM of three independent experiments. *, P < 0.05 vs. cells infected with Scr-shRNA lentivirus.
Mentions: We next explored the effects of PAK5 inhibition in glioma cells. First, we used a MTT assay to assess cell growth dynamics in Scr-shRNA- and PAK5-shRNA-infected cells. After 5 days, in U87 and SHG-44 cells, PAK5-shRNA-infected cells displayed remarkable cell growth inhibition compared with Scr-shRNA-infected cells. At day 5, the cell viability of the Scr-shRNA-infected group was 4 times higher than that of the PAK5-shRNA-infected groups (Fig. 3A and B). However, there was no significant difference between Scr-shRNA- and PAK5-shRNA-infected CHG-5 cells, cells with low PAK5 expression (Fig. 3C). To further determine the effect of PAK5 in glioma cells with high PAK5 expression, we enhanced PAK5 expression by transfecting PAK5 vector in PAK5-shRNA1-infected U87 cells (Fig. 3D). The results showed that enhancing PAK5 expression rescued the decrease of cell viability resulting from RNAi-mediated PAK5 knockdown (Fig. 3E).

Bottom Line: In addition, silencing PAK5 expression in U87 cells weakened their colony formation ability and in vivo tumorigenesis ability.Further studies demonstrated that PAK5 inhibition led to an increase in cleaved caspase 3 and a decrease in β-catenin.In conclusion, our results suggest that the inhibition of PAK5 by RNA interference might efficiently suppress tumor development of glioma cells with PAK5 high expression.

View Article: PubMed Central - PubMed

Affiliation: 1. Institute of Neurology, Guangdong Medical College, Zhanjiang, China. ; 2. Guangdong Key Laboratory of Age-Related Cardiac and Cerebral Diseases, Affiliated Hospital of Guangdong Medical College, Zhanjiang, China.

ABSTRACT
Glioma is the most common type of primary intracranial tumor and is highly lethal due to its pathogenetic location, high invasiveness, and poor prognosis. Even combined surgery and chemoradiotherapy do not effectively rescue glioma patients. Molecular target therapy is considered a safe and promising therapy for glioma. The identification of a novel, effective target protein in gliomas is of great interest. We found that PAK5 was highly expressed in the tumor tissues of glioma patients and human glioma cell lines. We then used a lentivirus-delivered short hairpin RNA to stably silence PAK5 expression in glioma cells and explore its influence. The results showed that the inhibition of PAK5 reduced cell viability and delayed the cell cycle at the G0/G1 phase in the glioma cells with PAK5 high expression. In addition, silencing PAK5 expression in U87 cells weakened their colony formation ability and in vivo tumorigenesis ability. Further studies demonstrated that PAK5 inhibition led to an increase in cleaved caspase 3 and a decrease in β-catenin. In conclusion, our results suggest that the inhibition of PAK5 by RNA interference might efficiently suppress tumor development of glioma cells with PAK5 high expression. This finding provides a novel, promising therapeutic target for glioma treatment.

Show MeSH
Related in: MedlinePlus