Helix pomatia agglutinin binding in human tumour cell lines: correlation with pulmonary metastases in nude mice

Abstract

Summary The extent of lectin binding by three human melanoma (LOX, FEMX-I and SESX) and two sarcoma lines (MHMX and OHSX) was related to their potential for experimental metastasis formation in athymic nude mice. The Helix pomatia agglutinin (HPA), which recognises the N-acetyl-D-galactosamine ligand, showed differential binding to the cell lines in a manner that correlated with their ability to give lung colonies after i.v. injection in the mice (P<0.005). The degree of HPA binding and lung colony formation of the cell lines studied was ranked in the following order, LOX> MHMX> OHSX> SESX > FEMX-I. Similar patterns were not observed with the other lectins used in this study (WGA, Con A, PNA and UEA-I). The high HPA reacting LOX melanoma line shows extensive pulmonary metastatic formation with no extrapulmonary colonies, whereas the low HPA reacting FEMX-I cells give only extrapulmonary metastases with no detectable colonies in the lungs. Precoating of tumour cells with HPA prior to injection did not reduce the ability of cells to give pulmonary metastases, suggesting that the HPA epitope was not functionally associated with the pulmonary metastatic potential observed in nude mice. These findings support recent human studies of a correlation between HPA binding and incidence of metastasis, however, our data indicate that there is no causal relationship. Further analyses are required to identify the specific HPA-binding glycoconjugates that may be involved.Cancer cells often display aberrant glycosylation of surface membrane proteins and lipids (Hakomori & Kannagi, 1983;Reading & Hutchins, 1985). The altered structure or expression of these sugar-containing molecules on malignant cells is thought to be important for some of the events leading to the formation of metastasis (Nicolson, 1984) (Leathem & Brooks, 1987;Springer, 1989;Brooks & Leathem, 1991;Fukutomi et al., 1991;Schumacher et al., 1992), gastrointestinal tract (Kakeji et al., 1991) and prostate (Shiraishi et al., 1992). Most experimental studies on the role of membrane glyconjugates in the metastatic process have been performed in syngeneic rodent tumour systems (Tao & Burger, 1977;Dennis et al., 1981;Finne et al., 1989;Hagmar et al., 1990). In the present study we have used human melanoma and sarcoma cell lines in nude mice to examine whether a relationship between lectin binding and metastatic potential could be detected. Materials and methods AnimalsCongenitally nude mice (Balb/c) were bred and maintained as previously described (Fodstad et al., 1988a). Animals (4-6 weeks of age) of both sexes were used. (Fodstad et al., 1986) and the MHMX unclassified sarcoma. The cell lines were all established from biopsies of patients at the Norwegian Radium Hospital. The cells were cultured as monolayers and as xenografts in nude mice. The monolayer cultures were maintained at 37°C in RPMI-1640 medium (Gibco, Paisley, UK) supplemented with 10% fetal calf serum (Serva, Heidelberg, Germany), 1,000 IU ml-' penicillin, 1I00 lg ml-' streptomycin and 2 mg I' L-glutam...

Clinically proven markers of metastasis predict metastatic spread of human melanoma cells engrafted in scid mice

“…Also, intensely HPA-positive and metastatic cell line MeWo shows a tumour growth phase of 90 days, which has to be taken into consideration, concerning therapeutic studies. Kjonniksen et al (1994) demonstrated that the metastatic cell line LOX showed strong HPA binding, which is in parallel to our results. Additional results by that group showed, that the HPAnegative cell line FEMX-1 was not metastatic after iv injection, which is in contrast to our results, where all primary FEMX-1 tumours expressed HPA-binding sites and developed metastatic deposits in the lungs of 7/10 mice.…”

“…Furthermore, subcutaneous injection of tumour cells, as presented here, warrants that the metastatic cascade as a whole can be analysed, whereas intravenous (iv) injection of tumour cells, as sometimes described by others, results in dissemination rather than in metastasis (Kjonniksen et al, 1994). For malignant melanoma, in particular, subcutaneous injection has the advantage of being an almost orthotopic model and therefore, reflects the clinical situation from the anatomical point of view more closely than the aforementioned iv injection.…”

“…We have established routine screening for mycoplasma, as about 30% of the permanent cell lines transferred to our facilities are mycoplasma infected. Earlier xenograft experiments with FEMX-1 and MeWo (data not shown) showed that both cell lines were Clinically relevant melanoma model A Thies et al indeed less tumorigenic, did not metastasise into the lungs and were HPA-negative, comparable to the results by Kjonniksen et al (1994). They further did not express CEACAM1 (MeWo) and/or L1 (MeWo, FEMX-1).…”

Human Tumor Antigens Tn and Sialyl Tn Arise from Mutations inCosmc

“…To examine the correlation in LOX cells between loss of Cosmc transcripts and Tn expression, we examined whether LOX cells were stained by HPA, as previously reported (23). LOX cells bound well to HPA and had poor binding to PNA.…”

Melanoma-specific expression in first-generation adenoviral vectors in vitro and in vivo — use of the human tyrosinase promoter with human enhancers

“…[13][14][15][16] The osteosarcoma cell line, OHS, was established from a bone tumor biopsy. 17 SK-MEL-28 (human melanoma), MeWo (human metastatic melanoma), A549 (human lung carcinoma), SK-BR-3 (human breast adenocarcinoma) and HeLa (human cervix epithelial adenocarcinoma) were obtained from the American Type Culture Collection (Rockville, MD).…”