Metastatic potential of human renal cell carcinoma: experimental model using subrenal capsule implantation in athymic nude mice

Abstract

Summary. The aim of this study was to determine whether subrenal capsule (SRC) implantation is a suitable model for the study of the metastatic potential of our human renal cell carcinoma (HRCC) lines and to establish new sublines with enhanced metastatic ability. NMRI athymic nude mice 7-11 weeks old received SRC implantation of our HRCC lines RC43 and RC21. These lines were not metastatic when implanted s.c. Mice were killed after 4 or 8 weeks, or when moribund. With the RC43 cell line the success rate for implantation was 69 %, with 89% of these showing metastases. The average volume of the implanted tumour fragments was 0.5 mm 3 (range 0.28-0.7), the average volume at the primary site at the time of death was 9087 (9-32000) mm 3. Metastases were found in lymph nodes, liver, spleen, peritoneum, psoas muscle, pancreas, diaphragm and skin. The average volume of the metastases was 4139 (0.5-9000) mm 3. Growing cell lines were established in vivo and in vitro from one splenic, one peritoneal, one diaphragmatic, and one hepatic metastasis. These sublines have faster in vivo and slower in vitro growth rates than the parental lines. With the RC21 cell line the success rate for implantation was 56% and the metastatic rate 78%. The average volume of the implanted tumour was 0.8 mm 3 (0.28-1.2), the average volume at the primary site at the time of death was 2685 mm 3 (1.4-6534) and the average volume of metastases was 7.1 mm 3 (0.5-37.5). Metastases were found in lymph nodes, lung and skin. No establishment was attempted for RC21 because of the small dimensions of these metastases. SRC implantation is thus considered a useful tool for the study of the metastatic ability of our cell lines RC43 and RC21. The establishment of new sublines with a faster growth rate and an enhanced metastatic ability will be useful for further studies on the metastatic process. Key words: Metastatic potential -Subrenal capsule implantation -Human Renal Cell CarcinomaCancer patients rarely die of their primary tumours. Establishment of metastases in various organs and their subsequent growth usually cause the terminal cachexia of neoplastic patients.The non-random pattern of visceral metastases suggested to Paget [1] more than 100 years ago that the growth of metastases was due to an affinity between turnout cells (seeds) and a suitable environment in some organs (soil). Since then, the metastatic process has only been partially clarified, mainly because of the relative scarcity of experimental models that could reliably reproduce the interactions between host and tumour cells.Human renal cell carcinoma (HRCC) is an uncommon malignancy (6 % of all tumours diagnosed each year) with a poor prognosis ranging from 73 to 8 % survival at 5 years depending on the stage at diagnosis [2]. The establishment of HRCC lines growing in vitro or in vivo is not easy. In our laboratory only 10 % of implanted tumours produced a serially transplantable line [3]. Furthermore, the simple model of an s.c.-growing tumour is not very useful for research regarding ...

Progression of renal cell carcinoma is inhibited by genistein and radiation in an orthotopic model

“…We showed that renal implantation of the KCI-18 cell line in the kidney of nude mice caused the formation of a primary kidney tumor that metastasizes to the lungs, liver and mesentery; mimicking the development and progression of metastatic RCC in human [2,3,21]. Our studies and others demonstrate that orthotopic models of heterotransplanted human RCC cells are representative of human RCC progression and preserve the karyotypic and histological characteristics of the human primary tumor [6-9,20]. These models can be used to study human RCC progression and metastasis and responsiveness of human RCC tumors to treatment in vivo .…”

“…Among others, tumor xenograft models established in immunodeficient mice by implantation of RCC cells, isolated from a human tumor specimen, have been valuable to assess responsiveness to therapy [6-9]. To investigate the combination of radiotherapy with other treatment modalities, we have established a xenograft metastatic RCC tumor model by orthotopic renal implantation of a new KCI-18 human RCC cell line in athymic nude mice.…”

Decreased levels of topoisomerase IIα in human renal cell carcinoma lines resistant to etoposide

“…Cell culture and conditions Two non-chemoselected human RCC cell lines, RC2 and RC21, originally established in our laboratory (Grossi et al 1992) and three resistant sublines were examined. All RCC cell lines were cultured in DMEM-F12 medium supplemented with 7.5% fetal calf serum, 1 E/ml penicillin, 1lg/ml streptomycin and 2 mM glutamine (Gibco BRL Europe, Breda, The Netherlands) at 37°C in a humidi®ed atmosphere supplemented with 5% CO 2 .…”

Characterization of 11 human sarcoma cell strains

“…28 Orthotopic sites that most closely resemble the microenvironments in which human tumors develop are the most likely sites for successful tumor implantation. 28 In fact, it has been observed that orthotopically implanted human tumor xenografts have an increased chance of establishment, growth, and metastasis in immunodeficient animals when compared with nonorthotopically implanted xenografts, 28 as is the case with human renal cell carcinoma 29 and breast carcinoma. 30 Additionally, in experiments using the subcutaneous implantation method, only the cell strain MFH-3 consistently developed tumor nodules at the injection site.…”