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Lentiviral Vector Mediated Claudin1 Silencing Inhibits Epithelial to Mesenchymal Transition in Breast Cancer Cells.

Zhao X, Zou Y, Gu Q, Zhao G, Gray H, Pfeffer LM, Yue J - Viruses (2015)

Bottom Line: Claudin1 (CLDN1) is a member of the tetraspan transmembrane protein family that plays a major role in tight junctions and is associated with tumor metastasis.We found that a CLDN1 short hairpin (shRNA) construct efficiently silenced CLDN1 expression in both breast cancer cell lines, and CLDN1 knockdown resulted in reduced cell proliferation, survival, migration and invasion.Furthermore, silencing CLDN1 inhibited epithelial to mesenchymal transition (EMT) by upregulating the epithelial cell marker, E-cadherin, and downregulating mesenchymal markers, smooth muscle cell alpha-actin (SMA) and Snai2.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Harbin Medical University, Harbin 150086, China. ynzou@sina.com.

ABSTRACT
Breast cancer has a high incidence and mortality rate worldwide. Several viral vectors including lentiviral, adenoviral and adeno-associated viral vectors have been used in gene therapy for various forms of human cancer, and have shown promising effects in controlling tumor development. Claudin1 (CLDN1) is a member of the tetraspan transmembrane protein family that plays a major role in tight junctions and is associated with tumor metastasis. However, the role of CLDN1 in breast cancer is largely unexplored. In this study, we tested the therapeutic potential of silencing CLDN1 expression in two breast cancer (MDA-MB-231 and MCF7) cell lines using lentiviral vector mediated RNA interference. We found that a CLDN1 short hairpin (shRNA) construct efficiently silenced CLDN1 expression in both breast cancer cell lines, and CLDN1 knockdown resulted in reduced cell proliferation, survival, migration and invasion. Furthermore, silencing CLDN1 inhibited epithelial to mesenchymal transition (EMT) by upregulating the epithelial cell marker, E-cadherin, and downregulating mesenchymal markers, smooth muscle cell alpha-actin (SMA) and Snai2. Our data demonstrated that lentiviral vector mediated CLDN1 RNA interference has great potential in breast cancer gene therapy by inhibiting EMT and controlling tumor cell growth.

No MeSH data available.


Related in: MedlinePlus

Silencing CLDN1 inhibits breast cancer cell survival. (A,B) Cell survival in MDA-MB-231 (A) and MCF7 (B) cells transduced with different CLDN1 lentiviral shRNAs and control vectors were examined by cell colony formation assays. Cell colonies were counted after culture (two weeks) in six-well plates and Crystal Violet staining. The number of colonies in CLDN1 lentiviral shRNA transduced cells was compared to that in control cells. Data were analyzed and presented from three independent experiments (** p < 0.01; *** p < 0.001).
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viruses-07-02755-f003: Silencing CLDN1 inhibits breast cancer cell survival. (A,B) Cell survival in MDA-MB-231 (A) and MCF7 (B) cells transduced with different CLDN1 lentiviral shRNAs and control vectors were examined by cell colony formation assays. Cell colonies were counted after culture (two weeks) in six-well plates and Crystal Violet staining. The number of colonies in CLDN1 lentiviral shRNA transduced cells was compared to that in control cells. Data were analyzed and presented from three independent experiments (** p < 0.01; *** p < 0.001).

Mentions: To examine whether CLDN1 affected breast cancer cell survival, we performed colony formation assays in both MDA-MB-231 and MCF7 cells transduced with CLDN1 lentiviral shRNAs and SC control vectors. Cell colonies were counted following a two-week culture period. Silencing CLDN1 significantly reduced cell survival in both MDA-MB-231 (Figure 3A) and MCF7 (Figure 3B) cells transduced with CLDN1 lentiviral shRNA vectors compared to SC transduced control cells.


Lentiviral Vector Mediated Claudin1 Silencing Inhibits Epithelial to Mesenchymal Transition in Breast Cancer Cells.

Zhao X, Zou Y, Gu Q, Zhao G, Gray H, Pfeffer LM, Yue J - Viruses (2015)

Silencing CLDN1 inhibits breast cancer cell survival. (A,B) Cell survival in MDA-MB-231 (A) and MCF7 (B) cells transduced with different CLDN1 lentiviral shRNAs and control vectors were examined by cell colony formation assays. Cell colonies were counted after culture (two weeks) in six-well plates and Crystal Violet staining. The number of colonies in CLDN1 lentiviral shRNA transduced cells was compared to that in control cells. Data were analyzed and presented from three independent experiments (** p < 0.01; *** p < 0.001).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4488722&req=5

viruses-07-02755-f003: Silencing CLDN1 inhibits breast cancer cell survival. (A,B) Cell survival in MDA-MB-231 (A) and MCF7 (B) cells transduced with different CLDN1 lentiviral shRNAs and control vectors were examined by cell colony formation assays. Cell colonies were counted after culture (two weeks) in six-well plates and Crystal Violet staining. The number of colonies in CLDN1 lentiviral shRNA transduced cells was compared to that in control cells. Data were analyzed and presented from three independent experiments (** p < 0.01; *** p < 0.001).
Mentions: To examine whether CLDN1 affected breast cancer cell survival, we performed colony formation assays in both MDA-MB-231 and MCF7 cells transduced with CLDN1 lentiviral shRNAs and SC control vectors. Cell colonies were counted following a two-week culture period. Silencing CLDN1 significantly reduced cell survival in both MDA-MB-231 (Figure 3A) and MCF7 (Figure 3B) cells transduced with CLDN1 lentiviral shRNA vectors compared to SC transduced control cells.

Bottom Line: Claudin1 (CLDN1) is a member of the tetraspan transmembrane protein family that plays a major role in tight junctions and is associated with tumor metastasis.We found that a CLDN1 short hairpin (shRNA) construct efficiently silenced CLDN1 expression in both breast cancer cell lines, and CLDN1 knockdown resulted in reduced cell proliferation, survival, migration and invasion.Furthermore, silencing CLDN1 inhibited epithelial to mesenchymal transition (EMT) by upregulating the epithelial cell marker, E-cadherin, and downregulating mesenchymal markers, smooth muscle cell alpha-actin (SMA) and Snai2.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Harbin Medical University, Harbin 150086, China. ynzou@sina.com.

ABSTRACT
Breast cancer has a high incidence and mortality rate worldwide. Several viral vectors including lentiviral, adenoviral and adeno-associated viral vectors have been used in gene therapy for various forms of human cancer, and have shown promising effects in controlling tumor development. Claudin1 (CLDN1) is a member of the tetraspan transmembrane protein family that plays a major role in tight junctions and is associated with tumor metastasis. However, the role of CLDN1 in breast cancer is largely unexplored. In this study, we tested the therapeutic potential of silencing CLDN1 expression in two breast cancer (MDA-MB-231 and MCF7) cell lines using lentiviral vector mediated RNA interference. We found that a CLDN1 short hairpin (shRNA) construct efficiently silenced CLDN1 expression in both breast cancer cell lines, and CLDN1 knockdown resulted in reduced cell proliferation, survival, migration and invasion. Furthermore, silencing CLDN1 inhibited epithelial to mesenchymal transition (EMT) by upregulating the epithelial cell marker, E-cadherin, and downregulating mesenchymal markers, smooth muscle cell alpha-actin (SMA) and Snai2. Our data demonstrated that lentiviral vector mediated CLDN1 RNA interference has great potential in breast cancer gene therapy by inhibiting EMT and controlling tumor cell growth.

No MeSH data available.


Related in: MedlinePlus