Limits...
Novel LIMK2 Inhibitor Blocks Panc-1 Tumor Growth in a mouse xenograft model.

Rak R, Haklai R, Elad-Tzfadia G, Wolfson HJ, Carmeli S, Kloog Y - Oncoscience (2014)

Bottom Line: We recently reported the development and validation of a novel LIMK inhibitor, referred to here as T56-LIMKi, using a combination of computational methods and classical biochemistry techniques.We found that T56-LIMKi decreases phosphorylated cofilin (p-cofilin) levels and thus inhibits growth of several cancerous cell lines, including those of pancreatic cancer, glioma and schwannoma.Because the most promising in-vitro effect of T56-LIMKi was observed in the pancreatic cancer cell line Panc-1, we tested the inhibitor on a nude mouse Panc-1 xenograft model.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurobiology, The George S. Wise Faculty of Life Sciences, Tel-Aviv University, Tel Aviv, Israel.

ABSTRACT
LIM kinases (LIMKs) are important cell cytoskeleton regulators that play a prominent role in cancer manifestation and neuronal diseases. The LIMK family consists of two homologues, LIMK1 and LIMK2, which differ from one another in expression profile, intercellular localization, and function. The main substrate of LIMK is cofilin, a member of the actin-depolymerizing factor (ADF) protein family. When phosphorylated by LIMK, cofilin is inactive. LIMKs play a contributory role in several neurodevelopmental disorders and in cancer growth and metastasis. We recently reported the development and validation of a novel LIMK inhibitor, referred to here as T56-LIMKi, using a combination of computational methods and classical biochemistry techniques. Here we report that T56-LIMKi inhibits LIMK2 with high specificity, and shows little or no cross-reactivity with LIMK1. We found that T56-LIMKi decreases phosphorylated cofilin (p-cofilin) levels and thus inhibits growth of several cancerous cell lines, including those of pancreatic cancer, glioma and schwannoma. Because the most promising in-vitro effect of T56-LIMKi was observed in the pancreatic cancer cell line Panc-1, we tested the inhibitor on a nude mouse Panc-1 xenograft model. T56-LIMKi reduced tumor size and p-cofilin levels in the Panc-1 tumors, leading us to propose T56-LIMKi as a candidate drug for cancer therapy.

No MeSH data available.


Related in: MedlinePlus

HeLa cells were stably transfected with the PC-vector or with HA-LIMK1 or HA-LIMK2 expression vector(A) Representative blots of the transfected cells obtained with anti-HA or anti-LIMK1 antibodies. (B) Results of the corresponding experiment using anti-LIMK2 antibodies.
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Figure 1: HeLa cells were stably transfected with the PC-vector or with HA-LIMK1 or HA-LIMK2 expression vector(A) Representative blots of the transfected cells obtained with anti-HA or anti-LIMK1 antibodies. (B) Results of the corresponding experiment using anti-LIMK2 antibodies.

Mentions: In our previous report using the NF1-depleted MEFs we were unable to tell whether T56-LIMKi inhibits LIMK1 or LIMK2 or both. To obtain this information, in the present work we performed in-vitro experiments with HeLa cells stably expressing the vehicle (control), LIMK1, or LIMK2. These cells were chosen because they express low levels of p-cofilin [41] and undergo transfection with high efficiency. We transfected HeLa cells with pcDNA3 vectors containing HA-tagged LIMK1 or LIMK2, or with an empty vector (Fig 1). We found that the cells transfected with HA-LIMK1 or HA-LIMK2 indeed expressed the corresponding enzymes (Fig. 1 A and B). The empty vector control exhibited only low basal levels of the LIMK1 and LIMK2 enzymes.


Novel LIMK2 Inhibitor Blocks Panc-1 Tumor Growth in a mouse xenograft model.

Rak R, Haklai R, Elad-Tzfadia G, Wolfson HJ, Carmeli S, Kloog Y - Oncoscience (2014)

HeLa cells were stably transfected with the PC-vector or with HA-LIMK1 or HA-LIMK2 expression vector(A) Representative blots of the transfected cells obtained with anti-HA or anti-LIMK1 antibodies. (B) Results of the corresponding experiment using anti-LIMK2 antibodies.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4295757&req=5

Figure 1: HeLa cells were stably transfected with the PC-vector or with HA-LIMK1 or HA-LIMK2 expression vector(A) Representative blots of the transfected cells obtained with anti-HA or anti-LIMK1 antibodies. (B) Results of the corresponding experiment using anti-LIMK2 antibodies.
Mentions: In our previous report using the NF1-depleted MEFs we were unable to tell whether T56-LIMKi inhibits LIMK1 or LIMK2 or both. To obtain this information, in the present work we performed in-vitro experiments with HeLa cells stably expressing the vehicle (control), LIMK1, or LIMK2. These cells were chosen because they express low levels of p-cofilin [41] and undergo transfection with high efficiency. We transfected HeLa cells with pcDNA3 vectors containing HA-tagged LIMK1 or LIMK2, or with an empty vector (Fig 1). We found that the cells transfected with HA-LIMK1 or HA-LIMK2 indeed expressed the corresponding enzymes (Fig. 1 A and B). The empty vector control exhibited only low basal levels of the LIMK1 and LIMK2 enzymes.

Bottom Line: We recently reported the development and validation of a novel LIMK inhibitor, referred to here as T56-LIMKi, using a combination of computational methods and classical biochemistry techniques.We found that T56-LIMKi decreases phosphorylated cofilin (p-cofilin) levels and thus inhibits growth of several cancerous cell lines, including those of pancreatic cancer, glioma and schwannoma.Because the most promising in-vitro effect of T56-LIMKi was observed in the pancreatic cancer cell line Panc-1, we tested the inhibitor on a nude mouse Panc-1 xenograft model.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurobiology, The George S. Wise Faculty of Life Sciences, Tel-Aviv University, Tel Aviv, Israel.

ABSTRACT
LIM kinases (LIMKs) are important cell cytoskeleton regulators that play a prominent role in cancer manifestation and neuronal diseases. The LIMK family consists of two homologues, LIMK1 and LIMK2, which differ from one another in expression profile, intercellular localization, and function. The main substrate of LIMK is cofilin, a member of the actin-depolymerizing factor (ADF) protein family. When phosphorylated by LIMK, cofilin is inactive. LIMKs play a contributory role in several neurodevelopmental disorders and in cancer growth and metastasis. We recently reported the development and validation of a novel LIMK inhibitor, referred to here as T56-LIMKi, using a combination of computational methods and classical biochemistry techniques. Here we report that T56-LIMKi inhibits LIMK2 with high specificity, and shows little or no cross-reactivity with LIMK1. We found that T56-LIMKi decreases phosphorylated cofilin (p-cofilin) levels and thus inhibits growth of several cancerous cell lines, including those of pancreatic cancer, glioma and schwannoma. Because the most promising in-vitro effect of T56-LIMKi was observed in the pancreatic cancer cell line Panc-1, we tested the inhibitor on a nude mouse Panc-1 xenograft model. T56-LIMKi reduced tumor size and p-cofilin levels in the Panc-1 tumors, leading us to propose T56-LIMKi as a candidate drug for cancer therapy.

No MeSH data available.


Related in: MedlinePlus