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Noncoding human Y RNAs are overexpressed in tumours and required for cell proliferation.

Christov CP, Trivier E, Krude T - Br. J. Cancer (2008)

Bottom Line: In particular, hY1 and hY3 RNAs are overexpressed in carcinomas (and adenocarcinomas) of the bladder, cervix, colon, kidney, lung and prostate with extremely high statistical significance (ANOVA, between groups, P<10e-22).A functional requirement of all four hY RNAs for cell proliferation was investigated in a systematic survey for loss-of-function by RNA interference (RNAi).Degradation of hY1 and hY3 RNAs in human cell lines resulted in a significant cytostatic inhibition of cell proliferation.

View Article: PubMed Central - PubMed

Affiliation: Department of Zoology, University of Cambridge, Downing Street, Cambridge CB2 3EJ, UK.

ABSTRACT
Noncoding Y RNAs have recently been identified as essential factors for chromosomal DNA replication in human cell nuclei. Here, we investigate the expression of human Y RNAs in tumours and test their requirement for cell proliferation. Relative expression levels of all four human Y RNAs (hY1, hY3, hY4 and hY5 RNA) were determined by quantitative RT-PCR in extracts from human solid tumours, corresponding nonmalignant normal tissues and derived cultured cells. On average, all four hY RNAs are significantly overexpressed in solid tumours between 4- and 13-fold, compared to the corresponding normal tissues. In particular, hY1 and hY3 RNAs are overexpressed in carcinomas (and adenocarcinomas) of the bladder, cervix, colon, kidney, lung and prostate with extremely high statistical significance (ANOVA, between groups, P<10e-22). A functional requirement of all four hY RNAs for cell proliferation was investigated in a systematic survey for loss-of-function by RNA interference (RNAi). Degradation of hY1 and hY3 RNAs in human cell lines resulted in a significant cytostatic inhibition of cell proliferation. We conclude that noncoding hY RNAs have potential both as new cancer biomarkers and as molecular targets for anti-proliferative intervention.

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Depletion of hY3 RNA is cytostatic and does not cause cell death. (A) Cell viability assay. RNAi was performed on WI38 lung fibroblasts and HeLa cervical carcinoma cells. At 0 h and 48 h post transfection with the indicated siRNAs, percentages of viable cells were determined by measuring exclusion of the dye, trypan blue. (B) Cell morphology. Representative phase contrast micrographs of cells are shown at 48 h after transfection. The mitotic index (%M) for each of these cell populations was measured by counting >800 cells per sample and it is indicated at the bottom left of each field. Scale bar, 10 μm.
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fig5: Depletion of hY3 RNA is cytostatic and does not cause cell death. (A) Cell viability assay. RNAi was performed on WI38 lung fibroblasts and HeLa cervical carcinoma cells. At 0 h and 48 h post transfection with the indicated siRNAs, percentages of viable cells were determined by measuring exclusion of the dye, trypan blue. (B) Cell morphology. Representative phase contrast micrographs of cells are shown at 48 h after transfection. The mitotic index (%M) for each of these cell populations was measured by counting >800 cells per sample and it is indicated at the bottom left of each field. Scale bar, 10 μm.

Mentions: In the next set of experiments, we investigated whether degradation of hY RNAs in proliferating human cells leads to an inhibition of cell proliferation. All four hY RNAs were expressed in several cell lines investigated (Supplementary Figure S1), in agreement with earlier reports (Hendrick et al, 1981; Pruijn et al, 1993). Our previous experiments have already established that RNA interference (RNAi) against hY1 RNAs is feasible in human cells, and we reported that degradation of hY1 RNA by two separate siRNAs results in a reduced proportion of replicating cells (Christov et al, 2006). We have therefore extended this analysis and conducted a systematic survey of RNAi on all four hY RNAs and analysed the consequences on cell proliferation (Figure 4) and cell viability (Figure 5).


Noncoding human Y RNAs are overexpressed in tumours and required for cell proliferation.

Christov CP, Trivier E, Krude T - Br. J. Cancer (2008)

Depletion of hY3 RNA is cytostatic and does not cause cell death. (A) Cell viability assay. RNAi was performed on WI38 lung fibroblasts and HeLa cervical carcinoma cells. At 0 h and 48 h post transfection with the indicated siRNAs, percentages of viable cells were determined by measuring exclusion of the dye, trypan blue. (B) Cell morphology. Representative phase contrast micrographs of cells are shown at 48 h after transfection. The mitotic index (%M) for each of these cell populations was measured by counting >800 cells per sample and it is indicated at the bottom left of each field. Scale bar, 10 μm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2266855&req=5

fig5: Depletion of hY3 RNA is cytostatic and does not cause cell death. (A) Cell viability assay. RNAi was performed on WI38 lung fibroblasts and HeLa cervical carcinoma cells. At 0 h and 48 h post transfection with the indicated siRNAs, percentages of viable cells were determined by measuring exclusion of the dye, trypan blue. (B) Cell morphology. Representative phase contrast micrographs of cells are shown at 48 h after transfection. The mitotic index (%M) for each of these cell populations was measured by counting >800 cells per sample and it is indicated at the bottom left of each field. Scale bar, 10 μm.
Mentions: In the next set of experiments, we investigated whether degradation of hY RNAs in proliferating human cells leads to an inhibition of cell proliferation. All four hY RNAs were expressed in several cell lines investigated (Supplementary Figure S1), in agreement with earlier reports (Hendrick et al, 1981; Pruijn et al, 1993). Our previous experiments have already established that RNA interference (RNAi) against hY1 RNAs is feasible in human cells, and we reported that degradation of hY1 RNA by two separate siRNAs results in a reduced proportion of replicating cells (Christov et al, 2006). We have therefore extended this analysis and conducted a systematic survey of RNAi on all four hY RNAs and analysed the consequences on cell proliferation (Figure 4) and cell viability (Figure 5).

Bottom Line: In particular, hY1 and hY3 RNAs are overexpressed in carcinomas (and adenocarcinomas) of the bladder, cervix, colon, kidney, lung and prostate with extremely high statistical significance (ANOVA, between groups, P<10e-22).A functional requirement of all four hY RNAs for cell proliferation was investigated in a systematic survey for loss-of-function by RNA interference (RNAi).Degradation of hY1 and hY3 RNAs in human cell lines resulted in a significant cytostatic inhibition of cell proliferation.

View Article: PubMed Central - PubMed

Affiliation: Department of Zoology, University of Cambridge, Downing Street, Cambridge CB2 3EJ, UK.

ABSTRACT
Noncoding Y RNAs have recently been identified as essential factors for chromosomal DNA replication in human cell nuclei. Here, we investigate the expression of human Y RNAs in tumours and test their requirement for cell proliferation. Relative expression levels of all four human Y RNAs (hY1, hY3, hY4 and hY5 RNA) were determined by quantitative RT-PCR in extracts from human solid tumours, corresponding nonmalignant normal tissues and derived cultured cells. On average, all four hY RNAs are significantly overexpressed in solid tumours between 4- and 13-fold, compared to the corresponding normal tissues. In particular, hY1 and hY3 RNAs are overexpressed in carcinomas (and adenocarcinomas) of the bladder, cervix, colon, kidney, lung and prostate with extremely high statistical significance (ANOVA, between groups, P<10e-22). A functional requirement of all four hY RNAs for cell proliferation was investigated in a systematic survey for loss-of-function by RNA interference (RNAi). Degradation of hY1 and hY3 RNAs in human cell lines resulted in a significant cytostatic inhibition of cell proliferation. We conclude that noncoding hY RNAs have potential both as new cancer biomarkers and as molecular targets for anti-proliferative intervention.

Show MeSH
Related in: MedlinePlus