Increased dietary levels of α-linoleic acid inhibit mammary tumor growth and metastasis.

OBJECTIVE: The aim of this study was to determine whether α-linolenic acid (ALA ω-3 fatty acid) enriched diet affects growth parameters when applied to a syngeneic model of mammary carcinoma. MATERIALS AND METHODS: BALB/c mice were divided and fed with: 1) a chia oil diet, rich in ALA or 2) a corn oil diet, rich in linoleic acid (LA ω-6 fatty acid). Mice were subcutaneously inoculated with a tumor cell line LM3, derived from a murine mammary adenocarcinoma. RESULTS: After 35 days, tumor incidence, weight, volume and metastasis number were lower in the ALA-fed mice, while tumor latency time was higher, and the release of pro-tumor metabolites derived from ω-6 fatty acids decreased in the tumor. Compared to the control group, a lower number of mitosis, a higher number of apoptotic bodies and higher T-lymphocyte infiltration were consistently observed in the ALA group. An ALA-rich diet decreased the estrogen receptor (ER) α expression, a recognized breast cancer promotor while showing an opposite effect on ERß in tumor lysates. CONCLUSION: These data support the anticancer effect of an ALA-enriched diet, which might be used as a dietary strategy in breast cancer prevention.

Characterization of a fibroblastoid mammary carcinoma cell line (LM2) originated from a mouse adenocarcinoma.

We established and characterized a new mammary tumor cell line, LM2, derived from M2 mammary adenocarcinoma which spontaneously appeared in a BALB/c female mouse. The LM2 cell line has been maintained in culture for more than 40 passages and grows as poorly differentiated elongated cells. Ultrastructural and immunocytochemistry analysis revealed characteristic features of adenocarcinoma. Cytogenetic studies showed that LM2 cells are fundamentally hypotetraploid. They express metalloproteinases (MMP) and show high levels of plasminogen activator type urokinase (uPA). They were sensitive to nitric oxide (NO)-mediated cytotoxicity when NO derived from an exogenous donor. In vivo, although LM2 cells were able to grow in the lungs, they could not metastasize to the same target organ from s.c. primary tumors. The LM2 mouse mammary adenocarcinoma cell line is a suitable model to examine different aspects of tumor biology, in particular those related to the different pathways involved in the metastatic cascade and in the cytotoxicity mediated by NO.

Characterization of F3II, a sarcomatoid mammary carcinoma cell line originated from a clonal subpopulation of a mouse adenocarcinoma.

We characterized a new mammary tumor cell line, F3II, previously established in vitro from a clonal subpopulation of the BALB/c transplantable mammary adenocarcinoma M3, moderately metastatic to lung. The F3II cell line has been passaged > 50 times. It has grown as elongated cells adherent to the bottom of the flask. Cytogenetic studies showed that F3II cultures were nearly triploid. Tumor cells expressed fibronectin and showed high levels of cell-surface urokinase, a key protease in invasion and metastasis. F3II cells grew as poorly differentiated, spindle-cell carcinoma tumors (sarcomatoid carcinomas) with a prominent local invasiveness, a high angiogenic response, and a 90-100% incidence of lung metastases when inoculated s.c. into syngeneic mice. Ultrastructural and immunocytochemical analysis revealed characteristic features of carcinomas. Our data suggest that F3II is less differentiated and more aggressive than the original tumor line, supporting the notion that mammary carcinomas are heterogeneous neoplasms and contain subpopulations with diverse biologic behavior. The F3II mouse mammary sarcomatoid carcinoma line is a suitable model to examine antiinvasive, antiangiogenic, and antimetastatic agents.

Receptors characterization of intraperitoneally N-nitroso-N-methylurea-induced mammary tumors in rats.

Mammary adenocarcinomas induces in female Sprague-Dawley rats by three intraperitoneal injections of N-nitroso-N-methylurea were studied in order to characterize their estrogen (ER), progesterone (PgR), prolactin (PRLR) and epidermal growth factor (EGFR) receptors. All samples evaluated showed the presence of ER and PgR in the cytosol fraction and PRLR amd EGFR in the membrane fraction. Q (fmol/mg) and K(d) (nM) values were as follows: ER, 56 +/- 11 and 0.5 +/- 0.1; PgR, 109 +/- 25 and 2.2 +/- 0.5 and PRLR, 335 +/- 75 and 0.5 +/- 0.2, respectively. In all tumors studied, two specific sites were found for EGFR, one with Q(1) = 22 +/- 9 and K(d1) = 0.6 +/- 0.3, and the other with Q(2) = 125 +/- 33 and K(d2) = 2.1 +/- 0.5. Receptor content was found to be independent of tumor histopathological variety. Displacement index (DI) with estradiol and tamoxifen of [I(3)H]E2-ER binding showed great heterogeneity, with values ranging from 0.01 to1.54. No correlation between ER content and DI values was found. Antiestrogenic binding sites were not found in the microsomal fraction of ten mammary tumors examined. Proliferation of this experimental mammary tumor may be regulated by a complex interaction of steroid and polypeptide hormones, as well as growth factors.

Heparin receptors in two murine mammary adenocarcinomas with different metastatic ability: relationship with growth inhibition.

Binding of heparin to primary cultured cells of two murine mammary adenocarcinomas with low (M3) and high (MM3) lung, metastatic capacity was determined. Heparin binding was rapid, specific and saturable. MM3 cells grown for 24 h in fetal calf serum (FCS)-free medium exhibited a higher number of binding sites for 3H-heparin [(11 +/- 1) x 10(5) sites per cell than M3 cells [(6.9 +/- 0.6) x 10(5) sites per cell]. However, when M3 cells were grown in the presence of 2% FCS, they showed less heparin binding sites [(3.5 +/- 0.4) x 10(5) sites per cell]. In contrast, dissociation constants were very similar for MM3 and M3 cells grown with or without FCS (Kd = 2-4 x 10(-9) M). Furthermore, heparin inhibited MM3 and M3 cell growth both in the absence or presence of FCS. Competition studies showed that chemically modified heparins lacking antiproliferative effect (O-desulfated; O/N-desulfated N-acetylated and N-desulfated heparins) were not able to inhibit 3H-heparin binding. N-desulfated N-acetylated heparin, which had partial antiproliferative effect, partially inhibited 3H-heparin binding, while heparin with a high antiproliferative activity inhibited more than 90% 3H-heparin binding. The antiproliferative effect of heparin and chemically modified heparins seems to be related to their binding ability to the cell membrane.