Transforming growth factor (TGF-B1) has been shown to upregulated (increased exp

Question

Transforming growth factor (TGF-B1) has been shown to upregulated (increased expression) in various forms of cancer. Madine-Darby Canine Kidney (MDCK) cells were cultured in the absence or presence of TGF-?1. In addition, TGF-?1 stimulated cells were treated with drug 'A. Both adherens junction formation as well as cell motility was assessed Control TGF-B1 TGF-1Drug 'A Control TGF-B1 TGF-?1 +Drug A Control TGF-?1 TGF-?1 +DrugA' ? (A) Cultured MDCK kidney cells were treated with control, transforming growth factor-B1 (TGF-B1), or TGF-B1+Drug 'A" for 24 hours. performed by labeling cells for E-cadherin (green) and nuclei (red). (B) Phase- contrast images of cultured cells treated with similar conditions as (A). Cel morphology indicates cells that cluster as a sheet (control) or disperse throughout the culture dish (TGF-?1). (C) Scratch assay of cells grown in the presence of either control or TGF-B1. Phase-contrast images were taken immediately following the formation of the cell-free zone (0 hour) and 24 hours following scratch formation (24 hour) was a. What might be the effect of drug 'A' on cell dispersion when added to TGF-B1 treated cells? Use the data to support your conclusions. b. What might be the effect of drug 'A' on cell motility when added to TGF- B1 treated cells? c, what effect might TGF-?1 have on RhoA activity? Use the data to support your conclusions.

Explanation / Answer

A) The epithelial to mesenchymal transition (EMT) is a fundamental process in tumor metastasis. The most notable hallmark of EMT is the decrease of E-cadherin expression and the proinflammatory cytokine TGF-?1 has been found to be one of the most potent activators of the experimentally induced EMT. In fig. (A) treatment of MDCK cells with TGF-?1 shows reduction of green fluorescence indicating the reduced expression of E-cadherin and restoration of the fluorescence when drug 'A' was also given which inicates that the drug 'A' impairs the activity of TGF-?1 in reducing the expression of E-cadherin or directly upregulates the expression of E-cadherin.

B) Drug 'A' will hinder the cell motility when added to TGF-?1 treated cells.

C) Ras homolog gene family, member A (RhoA) is a small GTPase protein in the Rho family which has a prominent regulatory role in the regulation of cytoskeletal dynamics, transcription, cell cycle progression and cell transformation, cell motility. According to the data given TGF-?1 will promote RhoA activity and thus in turn cytoskeletal rearrangement, another phenomenon having a pivotal role in EMT.

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