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Knockdown of LRP/LR Induces Apoptosis in Breast and Oesophageal Cancer Cells.

Khumalo T, Ferreira E, Jovanovic K, Veale RB, Weiss SF - PLoS ONE (2015)

Bottom Line: This knockdown of LRP/LR resulted in a significant decrease of viability in the breast and oesophageal cancer cells as determined by an MTT assay.Transfection of MDA-MB 231 cells with esiRNA-RPSA directed against a different region of the LRP mRNA had similar effects on LRP/LR expression and cell viability compared to siRNA-LAMR1, excluding an off-target effect of siRNA-LAMR1.These observations indicate that LRP/LR is crucial for the maintenance of cellular viability of breast and oesophageal cancer cells and recommend siRNA technology targeting LRP expression as a possible novel alternative technique for breast and oesophageal cancer treatment.

View Article: PubMed Central - PubMed

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

ABSTRACT
Cancer is a global burden due to high incidence and mortality rates and is ranked the second most diagnosed disease amongst non-communicable diseases in South Africa. A high expression level of the 37kDa/67kDa laminin receptor (LRP/LR) is one characteristic of cancer cells. This receptor is implicated in the pathogenesis of cancer cells by supporting tumor angiogenesis, metastasis and especially for this study, the evasion of apoptosis. In the current study, the role of LRP/LR on cellular viability of breast MCF-7, MDA-MB 231 and WHCO1 oesophageal cancer cells was investigated. Western blot analysis revealed that total LRP expression levels of MCF-7, MDA-MB 231 and WHCO1 were significantly downregulated by targeting LRP mRNA using siRNA-LAMR1. This knockdown of LRP/LR resulted in a significant decrease of viability in the breast and oesophageal cancer cells as determined by an MTT assay. Transfection of MDA-MB 231 cells with esiRNA-RPSA directed against a different region of the LRP mRNA had similar effects on LRP/LR expression and cell viability compared to siRNA-LAMR1, excluding an off-target effect of siRNA-LAMR1. This reduction in cellular viability is as a consequence of apoptosis induction as indicated by the exposure of the phosphatidylserine protein on the surface of breast MCF-7, MDA-MB 231 and oesophageal WHCO1 cancer cells, respectively, detected by an Annexin-V/FITC assay as well as nuclear morphological changes observed post-staining with Hoechst. These observations indicate that LRP/LR is crucial for the maintenance of cellular viability of breast and oesophageal cancer cells and recommend siRNA technology targeting LRP expression as a possible novel alternative technique for breast and oesophageal cancer treatment.

No MeSH data available.


Related in: MedlinePlus

The effect of siRNA-mediated downregulation of LRP expression on the cellular viability of WHCO1, MCF-7 and MDA-MB 231 cells.The viability of WHCO1, MCF-7 and MDA-MB 231 cells was analysed 72 h post-transfection using an MTT assay. siRNA-LAMR1 treated WHCO1, MCF-7 and MDA-MB 231 as well as esiRNA-RPSA treated MDA-MB 231 cells revealed a significant reduction in cellular viability compared to untreated cells set to 100%. 8mM PCA and siRNA-scr were used as positive and negative controls, respectively. The error bars represent the standard deviation (n = 3) and a significant difference (* p < 0.05, ** p < 0.01, *** p <0.001) between the untreated control and the treated samples is indicated by an asterisk.
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pone.0139584.g004: The effect of siRNA-mediated downregulation of LRP expression on the cellular viability of WHCO1, MCF-7 and MDA-MB 231 cells.The viability of WHCO1, MCF-7 and MDA-MB 231 cells was analysed 72 h post-transfection using an MTT assay. siRNA-LAMR1 treated WHCO1, MCF-7 and MDA-MB 231 as well as esiRNA-RPSA treated MDA-MB 231 cells revealed a significant reduction in cellular viability compared to untreated cells set to 100%. 8mM PCA and siRNA-scr were used as positive and negative controls, respectively. The error bars represent the standard deviation (n = 3) and a significant difference (* p < 0.05, ** p < 0.01, *** p <0.001) between the untreated control and the treated samples is indicated by an asterisk.

Mentions: In this study the MTT results suggest that LRP/LR plays a role in the viability of cancer cells. No significant decrease in viability of MDA-MB-231 was observed 24 h or 48 h after transfection (Fig 3), however the cell viability of the MCF-7, MDA-MB 231 and WHCO1 cancer cells was significantly decreased by 52%, 45% and 72%, respectively (Fig 4) 72 h following the downregulation of LRP/LR expression. 8mM PCA (a known apoptosis-inducer) was used as a positive control and cells transfected with the non-targeting siRNA-scr (or esiRNA-RLUC) posed as a negative control. A significant decrease in cellular viability was also noted in the 8mM PCA treated cells and the non-targeting siRNAs had no effect (Fig 4).


Knockdown of LRP/LR Induces Apoptosis in Breast and Oesophageal Cancer Cells.

Khumalo T, Ferreira E, Jovanovic K, Veale RB, Weiss SF - PLoS ONE (2015)

The effect of siRNA-mediated downregulation of LRP expression on the cellular viability of WHCO1, MCF-7 and MDA-MB 231 cells.The viability of WHCO1, MCF-7 and MDA-MB 231 cells was analysed 72 h post-transfection using an MTT assay. siRNA-LAMR1 treated WHCO1, MCF-7 and MDA-MB 231 as well as esiRNA-RPSA treated MDA-MB 231 cells revealed a significant reduction in cellular viability compared to untreated cells set to 100%. 8mM PCA and siRNA-scr were used as positive and negative controls, respectively. The error bars represent the standard deviation (n = 3) and a significant difference (* p < 0.05, ** p < 0.01, *** p <0.001) between the untreated control and the treated samples is indicated by an asterisk.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4591328&req=5

pone.0139584.g004: The effect of siRNA-mediated downregulation of LRP expression on the cellular viability of WHCO1, MCF-7 and MDA-MB 231 cells.The viability of WHCO1, MCF-7 and MDA-MB 231 cells was analysed 72 h post-transfection using an MTT assay. siRNA-LAMR1 treated WHCO1, MCF-7 and MDA-MB 231 as well as esiRNA-RPSA treated MDA-MB 231 cells revealed a significant reduction in cellular viability compared to untreated cells set to 100%. 8mM PCA and siRNA-scr were used as positive and negative controls, respectively. The error bars represent the standard deviation (n = 3) and a significant difference (* p < 0.05, ** p < 0.01, *** p <0.001) between the untreated control and the treated samples is indicated by an asterisk.
Mentions: In this study the MTT results suggest that LRP/LR plays a role in the viability of cancer cells. No significant decrease in viability of MDA-MB-231 was observed 24 h or 48 h after transfection (Fig 3), however the cell viability of the MCF-7, MDA-MB 231 and WHCO1 cancer cells was significantly decreased by 52%, 45% and 72%, respectively (Fig 4) 72 h following the downregulation of LRP/LR expression. 8mM PCA (a known apoptosis-inducer) was used as a positive control and cells transfected with the non-targeting siRNA-scr (or esiRNA-RLUC) posed as a negative control. A significant decrease in cellular viability was also noted in the 8mM PCA treated cells and the non-targeting siRNAs had no effect (Fig 4).

Bottom Line: This knockdown of LRP/LR resulted in a significant decrease of viability in the breast and oesophageal cancer cells as determined by an MTT assay.Transfection of MDA-MB 231 cells with esiRNA-RPSA directed against a different region of the LRP mRNA had similar effects on LRP/LR expression and cell viability compared to siRNA-LAMR1, excluding an off-target effect of siRNA-LAMR1.These observations indicate that LRP/LR is crucial for the maintenance of cellular viability of breast and oesophageal cancer cells and recommend siRNA technology targeting LRP expression as a possible novel alternative technique for breast and oesophageal cancer treatment.

View Article: PubMed Central - PubMed

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

ABSTRACT
Cancer is a global burden due to high incidence and mortality rates and is ranked the second most diagnosed disease amongst non-communicable diseases in South Africa. A high expression level of the 37kDa/67kDa laminin receptor (LRP/LR) is one characteristic of cancer cells. This receptor is implicated in the pathogenesis of cancer cells by supporting tumor angiogenesis, metastasis and especially for this study, the evasion of apoptosis. In the current study, the role of LRP/LR on cellular viability of breast MCF-7, MDA-MB 231 and WHCO1 oesophageal cancer cells was investigated. Western blot analysis revealed that total LRP expression levels of MCF-7, MDA-MB 231 and WHCO1 were significantly downregulated by targeting LRP mRNA using siRNA-LAMR1. This knockdown of LRP/LR resulted in a significant decrease of viability in the breast and oesophageal cancer cells as determined by an MTT assay. Transfection of MDA-MB 231 cells with esiRNA-RPSA directed against a different region of the LRP mRNA had similar effects on LRP/LR expression and cell viability compared to siRNA-LAMR1, excluding an off-target effect of siRNA-LAMR1. This reduction in cellular viability is as a consequence of apoptosis induction as indicated by the exposure of the phosphatidylserine protein on the surface of breast MCF-7, MDA-MB 231 and oesophageal WHCO1 cancer cells, respectively, detected by an Annexin-V/FITC assay as well as nuclear morphological changes observed post-staining with Hoechst. These observations indicate that LRP/LR is crucial for the maintenance of cellular viability of breast and oesophageal cancer cells and recommend siRNA technology targeting LRP expression as a possible novel alternative technique for breast and oesophageal cancer treatment.

No MeSH data available.


Related in: MedlinePlus