ABSTRACT
Suppression of detachment-induced cell death, known as anoikis, is an essential step for cancer metastasis to occur. We report here that expression of KLF12, a member of the Kruppel-like family of transcription factors, is downregulated in lung cancer cell lines that have been selected to grow in the absence of cell adhesion. Knockdown of KLF12 in parental cells results in decreased apoptosis following cell detachment from matrix. KLF12 regulates anoikis by promoting the cell cycle transition through S phase and therefore cell proliferation. Reduced expression levels of KLF12 results in increased ability of lung cancer cells to form tumours in vivo and is associated with poorer survival in lung cancer patients. We therefore identify KLF12 as a novel metastasis-suppressor gene whose loss of function is associated with anoikis resistance through control of the cell cycle.
Subject(s)
Anoikis/genetics , Gene Expression Regulation, Neoplastic , Kruppel-Like Transcription Factors/genetics , Lung Neoplasms/genetics , A549 Cells , Animals , Cell Cycle/genetics , Cell Line, Tumor , Cell Proliferation/genetics , Down-Regulation , Flow Cytometry , Humans , Lung Neoplasms/metabolism , Lung Neoplasms/pathology , Mice, Inbred NOD , Mice, Knockout , Mice, SCID , Neoplasm Metastasis , RNA Interference , Reverse Transcriptase Polymerase Chain Reaction , Survival Analysis , Transplantation, Heterologous , Tumor Suppressor Proteins/geneticsABSTRACT
The stiffness as well as the biodegradation rate of collagen and gelatine products can be modulated by performing a number of crosslinking treatments. In many biomedical applications, an optimal degree of crosslinking seems to exist, depending on the mechanical and/or biosynthesis properties of the host site. The aim of this study was to evaluate the optimal degree of crosslinking of collagen and gelatine films, to be used as sealants for vascular prostheses. Various crosslinking treatments, including exposure to aldehydes, dehydrothemal treatment, carbodiimide crosslinking and combinations of them, were performed on collagen and gelatine films, and the resulting increases in stiffness, degree of crosslinking and denaturation temperature were evaluated. Analogue crosslinking treatments were also performed on sealed prostheses, which were then tested for blood leakage. The experimental results showed that a good blood impermeability of both collagen and gelatine films was obtained for crosslinking density of about 1.2-1.3 x 10(-5) mol/cm(3), which could be yielded by a dehydrothermal crosslinking treatment (DHT). In particular, dehydrothermally treated gelatine-coated prostheses were found to perform better than analogue collagen-coated ones. The presence of glycerol in crosslinked collagen films was found to have plasticizing effects, which are likely to facilitate blood impermeability, and to increase the thermal stability of collagen.
