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Knock-Down of the 37kDa/67kDa Laminin Receptor LRP/LR Impedes Telomerase Activity.

Naidoo K, Malindisa ST, Otgaar TC, Bernert M, Da Costa Dias B, Ferreira E, Reusch U, Knackmuss S, Little M, Weiss SF, Letsolo BT - PLoS ONE (2015)

Bottom Line: FLAG® Co-immunoprecipitation assays confirmed an interaction between LRP/LR and hTERT.Knock-down of LRP/LR by RNAi technology significantly reduced telomerase activity.These results suggest for the first time a novel function of LRP/LR in contributing to telomerase activity. siRNAs targeting LRP/LR may act as a potential alternative therapeutic tool for cancer treatment by (i) blocking metastasis (ii) promoting angiogenesis (iii) inducing apoptosis and (iv) impeding telomerase activity.

View Article: PubMed Central - PubMed

Affiliation: School of Molecular and Cell Biology, University of the Witwatersrand, Private Bag 3, Wits, 2050, Johannesburg, Republic of South Africa.

ABSTRACT
Cancer has become a major problem worldwide due to its increasing incidence and mortality rates. Both the 37kDa/67kDa laminin receptor (LRP/LR) and telomerase are overexpressed in cancer cells. LRP/LR enhances the invasiveness of cancer cells thereby promoting metastasis, supporting angiogenesis and hampering apoptosis. An essential component of telomerase, hTERT is overexpressed in 85-90% of most cancers. hTERT expression and increased telomerase activity are associated with tumor progression. As LRP/LR and hTERT both play a role in cancer progression, we investigated a possible correlation between LRP/LR and telomerase. LRP/LR and hTERT co-localized in the perinuclear compartment of tumorigenic breast cancer (MDA_MB231) cells and non-tumorigenic human embryonic kidney (HEK293) cells. FLAG® Co-immunoprecipitation assays confirmed an interaction between LRP/LR and hTERT. In addition, flow cytometry revealed that both cell lines displayed high cell surface and intracellular LRP/LR and hTERT levels. Knock-down of LRP/LR by RNAi technology significantly reduced telomerase activity. These results suggest for the first time a novel function of LRP/LR in contributing to telomerase activity. siRNAs targeting LRP/LR may act as a potential alternative therapeutic tool for cancer treatment by (i) blocking metastasis (ii) promoting angiogenesis (iii) inducing apoptosis and (iv) impeding telomerase activity.

No MeSH data available.


Related in: MedlinePlus

Effect of LRP/LR on telomerase activity in HEK293 and MDA_MB231 cells.The expression level in HEK293 and MDA_MB231 cells was investigated using TRAPEZE Telomerase kit (Merck Millipore) and qPCR. Analysis of the concentrations revealed a significant (*** p < 0.001) reduction in telomerase activity once LRP was knockdown in A) HEK293, B) and C) MDA_MB231 cells, respectively, compared to control non-transfected cells and negative control siRNA transfected cells. Non-significant (ns) at p>0.05.
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pone.0141618.g005: Effect of LRP/LR on telomerase activity in HEK293 and MDA_MB231 cells.The expression level in HEK293 and MDA_MB231 cells was investigated using TRAPEZE Telomerase kit (Merck Millipore) and qPCR. Analysis of the concentrations revealed a significant (*** p < 0.001) reduction in telomerase activity once LRP was knockdown in A) HEK293, B) and C) MDA_MB231 cells, respectively, compared to control non-transfected cells and negative control siRNA transfected cells. Non-significant (ns) at p>0.05.

Mentions: The telomerase activity in response to the siRNA-mediated down regulation of LRP expression in HEK293 and MDA_MB231 cells was assessed using a TRAPeze RT® telomerase detection kit (Merck Millipore). HEK293 and MDA_MB231 cells were transfected with siRNA-LAMR1 and siRNA-TFRC/DICER1 (positive control). The level of telomerase activity in HEK293 and MDA_MB231 cells was significantly reduced (Fig 5) after the knock-down of LRP/LR.


Knock-Down of the 37kDa/67kDa Laminin Receptor LRP/LR Impedes Telomerase Activity.

Naidoo K, Malindisa ST, Otgaar TC, Bernert M, Da Costa Dias B, Ferreira E, Reusch U, Knackmuss S, Little M, Weiss SF, Letsolo BT - PLoS ONE (2015)

Effect of LRP/LR on telomerase activity in HEK293 and MDA_MB231 cells.The expression level in HEK293 and MDA_MB231 cells was investigated using TRAPEZE Telomerase kit (Merck Millipore) and qPCR. Analysis of the concentrations revealed a significant (*** p < 0.001) reduction in telomerase activity once LRP was knockdown in A) HEK293, B) and C) MDA_MB231 cells, respectively, compared to control non-transfected cells and negative control siRNA transfected cells. Non-significant (ns) at p>0.05.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4636255&req=5

pone.0141618.g005: Effect of LRP/LR on telomerase activity in HEK293 and MDA_MB231 cells.The expression level in HEK293 and MDA_MB231 cells was investigated using TRAPEZE Telomerase kit (Merck Millipore) and qPCR. Analysis of the concentrations revealed a significant (*** p < 0.001) reduction in telomerase activity once LRP was knockdown in A) HEK293, B) and C) MDA_MB231 cells, respectively, compared to control non-transfected cells and negative control siRNA transfected cells. Non-significant (ns) at p>0.05.
Mentions: The telomerase activity in response to the siRNA-mediated down regulation of LRP expression in HEK293 and MDA_MB231 cells was assessed using a TRAPeze RT® telomerase detection kit (Merck Millipore). HEK293 and MDA_MB231 cells were transfected with siRNA-LAMR1 and siRNA-TFRC/DICER1 (positive control). The level of telomerase activity in HEK293 and MDA_MB231 cells was significantly reduced (Fig 5) after the knock-down of LRP/LR.

Bottom Line: FLAG® Co-immunoprecipitation assays confirmed an interaction between LRP/LR and hTERT.Knock-down of LRP/LR by RNAi technology significantly reduced telomerase activity.These results suggest for the first time a novel function of LRP/LR in contributing to telomerase activity. siRNAs targeting LRP/LR may act as a potential alternative therapeutic tool for cancer treatment by (i) blocking metastasis (ii) promoting angiogenesis (iii) inducing apoptosis and (iv) impeding telomerase activity.

View Article: PubMed Central - PubMed

Affiliation: School of Molecular and Cell Biology, University of the Witwatersrand, Private Bag 3, Wits, 2050, Johannesburg, Republic of South Africa.

ABSTRACT
Cancer has become a major problem worldwide due to its increasing incidence and mortality rates. Both the 37kDa/67kDa laminin receptor (LRP/LR) and telomerase are overexpressed in cancer cells. LRP/LR enhances the invasiveness of cancer cells thereby promoting metastasis, supporting angiogenesis and hampering apoptosis. An essential component of telomerase, hTERT is overexpressed in 85-90% of most cancers. hTERT expression and increased telomerase activity are associated with tumor progression. As LRP/LR and hTERT both play a role in cancer progression, we investigated a possible correlation between LRP/LR and telomerase. LRP/LR and hTERT co-localized in the perinuclear compartment of tumorigenic breast cancer (MDA_MB231) cells and non-tumorigenic human embryonic kidney (HEK293) cells. FLAG® Co-immunoprecipitation assays confirmed an interaction between LRP/LR and hTERT. In addition, flow cytometry revealed that both cell lines displayed high cell surface and intracellular LRP/LR and hTERT levels. Knock-down of LRP/LR by RNAi technology significantly reduced telomerase activity. These results suggest for the first time a novel function of LRP/LR in contributing to telomerase activity. siRNAs targeting LRP/LR may act as a potential alternative therapeutic tool for cancer treatment by (i) blocking metastasis (ii) promoting angiogenesis (iii) inducing apoptosis and (iv) impeding telomerase activity.

No MeSH data available.


Related in: MedlinePlus