Murine precision-cut liver slices (PCLS): a new tool for studying tumor microenvironments and cell signaling ex vivo.
BACKGROUND
One of the most insidious characteristics of cancer is its spread to and ability to compromise distant organs via the complex process of metastasis. Communication between cancer cells and organ-resident cells via cytokines/chemokines and direct cell-cell contacts are key steps for survival, proliferation and invasion of metastasized cancer cells in organs. Precision-cut liver slices (PCLS) are considered to closely reflect the in vivo situation and are potentially useful for studying the interaction of cancer cells with liver-resident cells as well as being a potentially useful tool for screening anti-cancer reagents. Application of the PCLS technique in the field of cancer research however, has not yet been well developed.


RESULTS
We established the mouse PCLS system using perfluorodecalin (PFD) as an artificial oxygen carrier. Using this system we show that the adherence of green fluorescent protein (GFP) labeled MDA-MB-231 (highly invasive) cells to liver tissue in the PCLS was 5-fold greater than that of SK-BR-3 (less invasive) cells. In addition, we generated PCLS from THOC5, a member of transcription/export complex (TREX), knockout (KO) mice. The PCLS still expressed Gapdh or Albumin mRNAs at normal levels, while several chemokine/growth factor or metalloprotease genes, such as Cxcl12, Pdgfa, Tgfb, Wnt11, and Mmp1a genes were downregulated more than 2-fold. Interestingly, adhesion of cancer cells to THOC5 KO liver slices was far less (greater than 80% reduction) than to wild-type liver slices.


CONCLUSIONS
Mouse PCLS cultures in the presence of PFD may serve as a useful tool for screening local adherence and invasiveness of individual cancer cells, since single cells can be observed. This method may also prove useful for identification of genes in liver-resident cells that support cancer invasion by using PCLS from transgenic liver.
Institut fuer Biochemie, OE4310, Medizinische Hochschule Hannover, Carl-Neuberg-Str. 1, D-30623, Hannover, Germany. koch.alexandra@MH-Hannover.de.
Alexandra
A
Koch
Institut fuer Biochemie, OE4310, Medizinische Hochschule Hannover, Carl-Neuberg-Str. 1, D-30623, Hannover, Germany. saran.shashank@MH-Hannover.de.
Shashank
S
Saran
Institut fuer Biochemie, OE4310, Medizinische Hochschule Hannover, Carl-Neuberg-Str. 1, D-30623, Hannover, Germany. Tran.Doan@MH-Hannover.de.
Doan Duy Hai
DD
Tran
Institut fuer Biochemie, OE4310, Medizinische Hochschule Hannover, Carl-Neuberg-Str. 1, D-30623, Hannover, Germany. Klebba-Faerber.Sabine@MH-Hannover.de.
Sabine
S
Klebba-Färber
Institut fuer Biochemie, OE4310, Medizinische Hochschule Hannover, Carl-Neuberg-Str. 1, D-30623, Hannover, Germany. Hauke_Thiesler@web.de.
Hauke
H
Thiesler
Fraunhofer Institut für Toxikologie und Experimentelle Medizin Atemwegspharmakologie, Nikolai-Fuchs-Str.1, D-30625, Hannover, Germany. Katherina.sewald@item.fraunhofer.de.
Katherina
K
Sewald
Fraunhofer Institut für Toxikologie und Experimentelle Medizin Atemwegspharmakologie, Nikolai-Fuchs-Str.1, D-30625, Hannover, Germany. Susann.Schindler@item.fraunhofer.de.
Susann
S
Schindler
Fraunhofer Institut für Toxikologie und Experimentelle Medizin Atemwegspharmakologie, Nikolai-Fuchs-Str.1, D-30625, Hannover, Germany. Armin.braun@item.fraunhofer.de.
Armin
A
Braun
Institute of Veterinary Pathology, Freie Universitaet Berlin, Robert-von-Ostertag- Str. 15, D-14163, Berlin, Germany. robert.klopfleisch@fu-berlin.de.
Robert
R
Klopfleisch
Institut fuer Biochemie, OE4310, Medizinische Hochschule Hannover, Carl-Neuberg-Str. 1, D-30623, Hannover, Germany. tamura.teruko@MH-Hannover.de.
Teruko
T
Tamura
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D005466:Fluorocarbons; D049452:Green Fluorescent Proteins; D006801:Humans; D008099:Liver / Q000378:metabolism*; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009361:Neoplasm Invasiveness; D009369:Neoplasms / Q000378:metabolism / Q000473:pathology*; D017208:Rats, Wistar; D015398:Signal Transduction; D014280:Triglycerides / Q000378:metabolism; D059016:Tumor Microenvironment
D005466:Fluorocarbons; D014280:Triglycerides; D049452:Green Fluorescent Proteins; C008805:perfluorodecalin; D000255:Adenosine Triphosphate