Rat colon carcinoma cells that survived systemic immune surveillance are less sensitive to NK-cell mediated apoptosis

Abstract

In order to form distant metastases, cells from the primary tumor have to detach, enter the blood- or lymph-compartment and escape immune surveillance. Here, we describe the selection of rat colon carcinoma cell lines (CC531s-m1 and CC531s-m2) that escaped from systemic immune surveillance; CC531s cells were injected into the v. jugularis of Wag/Rij rats, after three weeks the lung tumors were isolated, the tumor cells were cultured, characterized and injected again. The m1- and m2-cell lines were less susceptible for killing by syngeneic NK cells. Further characterization of this cell line showed a decreased sensitivity towards TRAIL- and CD95L-, but not to granzyme B-mediated apoptosis. In the m1- and m2-cells log-phase growth started earlier as compared to the parental cell line, whereas no changes were found in anchorage-dependent or anchorage-independent growth. After subcapsular injection of the m2-cell line into the liver of rats much more lung metastases were formed in comparison to injection of the parental cell line. In conclusion, the results suggest that the resistance of the m1- and m2-cells to NK cell-mediated apoptosis was associated with their capability to survive systemic immune surveillance and form metastases in vivo.

Disrupted Expression of CXCL5 in Colorectal Cancer Is Associated with Rapid Tumor Formation in Rats and Poor Prognosis in Patients

“…The rat colon carcinoma cell line CC531S was originally developed using dimethylhydrazin in Wag/Rij rats (5). The aggressive CC531M was isolated from CC531S using an in vivo selection protocol (4,16). Cells were cultured at 37jC and 5% CO 2 , in cell culture flasks (Corning) containing culture medium, composed of RPMI1640 (Life Technologies), supplemented with 10% heat-inactivated FCS, 100 Ag/mL streptomycin, 100 IU/mL penicillin, and 2 mmol/L L-glutamine (all Life Technologies).…”

“…In a previous paper, we reported the in vivo selection of an aggressive rat colorectal cell line (CC531M) from the welldescribed CC531S cell line (4,5). The present study was initiated to identify factors that contribute to rapid growth and metastatic capacity of CC531M.…”

An improved method to study NK-independent mechanisms of MTLn3 breast cancer lung metastasis

“…In comparison, implantation of 1 · 10 6 MTLn3 cells in nude mice resulted in 26 lung metastases [5]. Also, intravenous injection of 2 · 10 6 selected, highly metastatic CC531-m2 tumor cells resulted only in several hundred experimental metastases in fully immunocompetent animals [18]. Thus 130 lung metastases after inoculation of no more than 1 · 10 5 MTLn3 cells in NK-cell depleted animals can be regarded as efficient and fully dependent on the lack of NK-cells as in non-treated animals almost no metastases were seen, even after injection of 1 · 10 6 cells.…”

The Tumor Microenvironment: A Pitch for Multiple Players

“…For example, several reports have elucidated the role of NK cells in killing cancer cells in murine models of melanoma, colon cancer, lung cancer, and breast cancer (Azogui et al, 1991; Pham-Nguyen et al, 1999; Velthuis et al, 2003; Carrega et al, 2009; Frings et al, 2011; Kim et al, 2011; Takeda et al, 2011; Vesely et al, 2011; Roberti et al, 2012a; Srivastava et al, 2012). There are several NK receptors, such as DNAM-1, CD155, CD16, CD69, NKp30, and NKp46, whose surface expression is fundamental for maintaining cancer immunosurveillance (Clausen et al, 2003; Garcia-Iglesias et al, 2009; Lakshmikanth et al, 2009; Chan et al, 2010; Levy et al, 2011; Pfeiffer et al, 2011; Gleason et al, 2012; Park et al, 2012a).…”