We have characterized the LCC15-MB cell line which was recently derived from a breast carcinoma metastasis resected from the femur of a 29-year-old woman. LCC15-MB cells are vimentin (VIM) positive, exhibit a stellate morphology in routine cell culture, and form penetrating colonies when embedded in three-dimensional gels of Matrigel or fibrillar collagen. They show high levels of activity in the Boyden chamber chemomigration and chemoinvasion assays, and like other invasive human breast cancer (HBC) cell lines, LCC15-MB cells activate matrix-metalloproteinase-2 in response to treatment with concanavalin A. In addition, these cells are tumorigenic when implanted subcutaneously in nude mice and recolonize bone after arterial injection. Interestingly, both the primary lesion and the bone metastasis from which LCC15-MB were derived, as well as the resultant cell line, abundantly express the bone matrix protein osteopontin (OPN). OPN is also expressed by the highly metastatic MDA-MB-435 cells, but not other invasive or noninvasive HBC cell lines. Expression of OPN is retained in the subcutaneous xenograft and intraosseous metastases of LCC15-MB as detected by immunohistochemistry. Both VIM and OPN expression have been associated with breast cancer invasion and metastasis, and their expression by the LCC15-MB cell line is consistent with its derivation from a highly aggressive breast cancer. These cells provide a useful model for studying molecular mechanisms important for breast cancer metastasis to bone and, in particular, the implication(s) of OPN and VIM expression in this process.
Bibliographical noteFunding Information:
Parts of the work were supported by the NIH Grant CA61344, the US Army Medical Research Acquisition Activity (USAMRAA) Grant DAMD17-96-1-6134, the SPORE in breast cancer NIH Grant 2P50-CA58185-04, and the Victorian Breast Cancer Research Consortium. We thank Drs. Larry Fisher and John Stubbs (NIDR, NIH) for their generous gifts of the OPN cDNA and antibodies and the UMR-106-01BSP cell line, Drs. Hynda Kleinman and Steve Akiyama (NIDR, NIH) for the matrix proteins, and Dr. John Newby, Washington County Hospital, Hagerstown, for providing original archival blocks. We also acknowledge the Lombardi Cancer Center Shared Resources FIG. 8. (A) A representative growth curve of subcutaneously implanted LCC15-MB xenografts over a period of 4 weeks. There were a total of ®ve animals per group and the experiment was repeated at least twice. (B) Frequency of organ-speci®c metastasis from a representative LCC15-MB cell intracardiac injection experiment as measured by resultant cell cultures (total number established) obtained following sacri®ce of the animal. Note that the majority of cultures, representing metastases (70%), arose in bony sites (femur, tibia, vertebra). There were a total of four animals per injection group.