Cells from a LPB cell line, a methylcholanthrene-induced C57Bl/6 mouse sarcoma cell line [27], were cultured using standard procedures in MEM (Gibco BRL, Cer-gy-Pontoise, France) supplemented with 100 U.ml−1 penicillin, 100 mg.ml−1 streptomycin (Sarbach, France) and 8% foetal calf serum (Gibco).
The S-180 mouse sarcoma cell line was obtained from the China Cell Culture Centre, grown and maintained in RPMI-1640 medium (Gibco) supplemented with 10% FBS.
Further work by other researchers in the 1980s supported this theory with studies of metastatic subclones from a mouse sarcoma cell line [ 26].
The mouse sarcoma cell line S-180 and macrophage cell line RAW264.7 were cultured in Eagle's Minimum Essential Medium and Dulbecco's modified Eagle's medium supplemented with 10% heat-inactivated fetal bovine serum, respectively.
The LPB cell line, a methylcholanthrene-induced C57Bl/6 mouse sarcoma cell line (Belehradek et al, 1972), and the B16 F0 melanoma cells (ATCC CRL 6322) were cultured using classical procedures and minimum essential medium culture medium (Gibco BRL, Cergy-Pontoise, France) supplemented with 100 U ml−1 penicillin, 100 mg ml−1 streptomycin (Sarbach, France) and 8% fetal calf serum (Gibco).
Mouse sarcoma cell lines were established from 2 Kras;p16p19 null mouse RMS tumors (T14-R, SMP-01) and one Kras;p16p19 null mouse NMS tumor (Sca1-01).
For the Kd studies using flow cytometry, mouse sarcoma cell lines engineered to express full-length human SAIL were incubated with increasing concentrations of antibodies.
Restoration of these downregulated miRNAs in mouse primary sarcoma cell lines showed that miR-16, but not other downregulated miRNAs, was able to significantly suppress both migration and invasion in vitro, without altering cell proliferation.
Restoration of miR-16 expression in mouse primary sarcoma cell lines significantly suppressed both migration and invasion in vitro and metastasis to the lung of sarcoma cells transplanted into the muscle of immunocompromised mice (an orthotopic mouse model of STS).
Experiments were performed using the mouse mammary sarcoma cell line, EMT-6 (obtained originally from Dr R Sutherland, University of Rochester, Rochester, NY, USA) and the human mammary carcinoma, MCF-7 obtained from American Type Culture Collection (Rockville, MD, USA).
