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Highly expressed ribosomal protein L34 indicates poor prognosis in osteosarcoma and its knockdown suppresses osteosarcoma proliferation probably through translational control

View Article: PubMed Central - PubMed

ABSTRACT

Osteosarcoma has devastating health implications on children and adolescents. However, due to its low incidence and high tumor heterogeneity, it is hard to achieve any further improvements in therapy and overall survival. Ribosomal protein L34 (RPL34) has been increasingly recognized to promote the proliferation of malignant cells, but its role in osteosarcoma has not been investigated. In this study, real-time quantitative PCR (RT-qPCR) and immunohistochemistry revealed that RPL34 was highly expressed in osteosarcoma tissues when compared to adjacent tissues and normal bone tissues. Survival analysis showed that high expression of RPL34 predicted a poor prognosis for osteosarcoma patients. Knockdown of RPL34 in Saos-2 cells via lentivirus-mediated small interfering RNA (siRNA) significantly inhibited cell proliferation, induced cell apoptosis and G2/M phase arrest. Moreover, screening of transcription factors using University of California Santa Cruz (UCSC) Genome Browser, protein-protein interaction (PPI) network analysis, Gene Ontology (GO) and pathway enrichment analysis revealed that MYC participates in the transcriptional regulation of RPL34, which interacts with the subunits of eukaryotic translation initiation factor 3 (eIF3) and probably involves the translational control of growth-promoting proteins. Our findings suggest that RPL34 plays an important role in the proliferation of osteosarcoma cells.

No MeSH data available.


RPL34 mRNA levels in 11 pairs of human OS tissues and adjacent tissues.(A) The interleaved bars show the fold changes of RPL34 mRNA levels in the OS tissues, as compared to those in the adjacent tissues. (B) The scatter plot displays a significant up-regulation of RPL34 mRNA in the OS tissues compared with the adjacent tissues, where the longer lines represent the median with interquartile range of 2−ΔΔCt, *P = 0.015.
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f1: RPL34 mRNA levels in 11 pairs of human OS tissues and adjacent tissues.(A) The interleaved bars show the fold changes of RPL34 mRNA levels in the OS tissues, as compared to those in the adjacent tissues. (B) The scatter plot displays a significant up-regulation of RPL34 mRNA in the OS tissues compared with the adjacent tissues, where the longer lines represent the median with interquartile range of 2−ΔΔCt, *P = 0.015.

Mentions: To investigate the effect of RPL34 expression on OS, we first determined the transcription level of RPL34 in human OS tissues and adjacent tissues by RT-qPCR, where RPL34 mRNA was shown to be up-regulated in 7/11 (63.64%) of the OS tissues at more than 3-fold higher levels (Fig. 1A). Compared with the adjacent tissues, the mRNA level of RPL34 was significantly increased in the OS tissues (Fig. 1B, P = 0.015). To confirm this result, RPL34 expression was further evaluated in 95 OS tissues and 60 normal bone tissues by immunohistochemistry analysis. Our data showed that the expression of RPL34 protein was detected in cytoplasm. According to the category standard, 75/95 (78.95%) of the OS tissues showed strongly positive staining with immunoreactive scores ranged from more than 4 to 12 and the other 20/95 (21.05%) OS tissues showed weakly positive staining with immunoreactive scores ranged from more than 0 to 4, whereas all of the normal bone tissues were weakly stained. The results revealed that RPL34 expression was also significantly up-regulated in the OS tissues, compared to the normal bone tissues (Fig. 2, P = 0.000).


Highly expressed ribosomal protein L34 indicates poor prognosis in osteosarcoma and its knockdown suppresses osteosarcoma proliferation probably through translational control
RPL34 mRNA levels in 11 pairs of human OS tissues and adjacent tissues.(A) The interleaved bars show the fold changes of RPL34 mRNA levels in the OS tissues, as compared to those in the adjacent tissues. (B) The scatter plot displays a significant up-regulation of RPL34 mRNA in the OS tissues compared with the adjacent tissues, where the longer lines represent the median with interquartile range of 2−ΔΔCt, *P = 0.015.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5121591&req=5

f1: RPL34 mRNA levels in 11 pairs of human OS tissues and adjacent tissues.(A) The interleaved bars show the fold changes of RPL34 mRNA levels in the OS tissues, as compared to those in the adjacent tissues. (B) The scatter plot displays a significant up-regulation of RPL34 mRNA in the OS tissues compared with the adjacent tissues, where the longer lines represent the median with interquartile range of 2−ΔΔCt, *P = 0.015.
Mentions: To investigate the effect of RPL34 expression on OS, we first determined the transcription level of RPL34 in human OS tissues and adjacent tissues by RT-qPCR, where RPL34 mRNA was shown to be up-regulated in 7/11 (63.64%) of the OS tissues at more than 3-fold higher levels (Fig. 1A). Compared with the adjacent tissues, the mRNA level of RPL34 was significantly increased in the OS tissues (Fig. 1B, P = 0.015). To confirm this result, RPL34 expression was further evaluated in 95 OS tissues and 60 normal bone tissues by immunohistochemistry analysis. Our data showed that the expression of RPL34 protein was detected in cytoplasm. According to the category standard, 75/95 (78.95%) of the OS tissues showed strongly positive staining with immunoreactive scores ranged from more than 4 to 12 and the other 20/95 (21.05%) OS tissues showed weakly positive staining with immunoreactive scores ranged from more than 0 to 4, whereas all of the normal bone tissues were weakly stained. The results revealed that RPL34 expression was also significantly up-regulated in the OS tissues, compared to the normal bone tissues (Fig. 2, P = 0.000).

View Article: PubMed Central - PubMed

ABSTRACT

Osteosarcoma has devastating health implications on children and adolescents. However, due to its low incidence and high tumor heterogeneity, it is hard to achieve any further improvements in therapy and overall survival. Ribosomal protein L34 (RPL34) has been increasingly recognized to promote the proliferation of malignant cells, but its role in osteosarcoma has not been investigated. In this study, real-time quantitative PCR (RT-qPCR) and immunohistochemistry revealed that RPL34 was highly expressed in osteosarcoma tissues when compared to adjacent tissues and normal bone tissues. Survival analysis showed that high expression of RPL34 predicted a poor prognosis for osteosarcoma patients. Knockdown of RPL34 in Saos-2 cells via lentivirus-mediated small interfering RNA (siRNA) significantly inhibited cell proliferation, induced cell apoptosis and G2/M phase arrest. Moreover, screening of transcription factors using University of California Santa Cruz (UCSC) Genome Browser, protein-protein interaction (PPI) network analysis, Gene Ontology (GO) and pathway enrichment analysis revealed that MYC participates in the transcriptional regulation of RPL34, which interacts with the subunits of eukaryotic translation initiation factor 3 (eIF3) and probably involves the translational control of growth-promoting proteins. Our findings suggest that RPL34 plays an important role in the proliferation of osteosarcoma cells.

No MeSH data available.