Chemistry: molecular biology and microbiology – Animal cell – per se ; composition thereof; process of... – Rodent cell – per se
Reexamination Certificate
2000-07-11
2004-10-26
Helms, Larry R. (Department: 1642)
Chemistry: molecular biology and microbiology
Animal cell, per se ; composition thereof; process of...
Rodent cell, per se
C435S325000
Reexamination Certificate
active
06808924
ABSTRACT:
FIELD OF THE INVENTION
The present invention relates to mouse mammary adenocarcinoma cell lines expressing estrogen and progesterone receptors as a tool to study the effect of hormones, pharmacological compounds, and environmental agents. The present invention also relates to methods in vitro and in vivo for testing hormones or other related molecules by cell proliferation or tumor proliferation.
BACKGROUND OF THE INVENTION
Epithelial breast malignant neoplasms of the mammary gland is one of the most common form of cancer among women in North America, South America, Europe, and Australia, and they account for 15-18% of the deaths in this population (Lynn and Reiss, 1995 J. Natl Cancer Inst. 87: 867; Moller Jensen et al., 1990 Eur. J. Cancer 26: 1167-1256; Boring et al., 1993 Cancer J. Clin. 43: 7-26). Despite the development of different therapeutic approaches, the mortality has continued to rise over the past thirty years: the incidence of breast cancer is increasing by about 1% per year in almost all populations in both industrialized and developing countries (Miller et al., 1991 Cancer Causes Control 2:67-74; Levi F., 1993 Eur J Cancer 29:2315-2319), and it is estimated that the disease will affect 5 million women in the next decade (Corry J F. and Lonning P E., 1994 Pharmacol. Econom 5: 198-212). Obviously, within the aging female population, prevention and treatment of breast cancer will continue to represent a major challenge.
The etiology of breast cancer remains largely unknown, and the dilemma of mammary tumor development and the related mechanisms have been studied with different experimental approaches. The virus-induced mouse mammary tumor (MMTV) model had different drawbacks; most of the tumors induced by MMTV are not hormone responsive or are pregnancy-responsive (Welsch C W and Nagasawa H., 1977 Cancer Res 37:951-963; Sluyser M. and Van Nie R., 1974 Cancer Res 34:3253-3257), and no definite involvement was demonstrated for a virus in the etiology of breast tumors in humans (Michalides R. et al., Current Topics in Microbiology and Immunology, Vol. 106, Eds: P K Vogt y H Koprowski, Springer Verlag, Berlin, pp57-78; Pogo B G. et al., 1997 Medicina (Buenos Aires)57 Suppl 2:75-80). The chemical carcinogen models allowed the dissection of initiators and promoters; the tumors originated in both the MNU and the DMBA rat models were hormone-responsive, and they expressed estrogen receptors (ER) (Gullino P. et al., 1975 J Natl Cancer Inst. 54:401-404; Russo I H. et al., 1982 Breast Cancer Res and Treat 2:5-73). They did not, however, give rise to metastasis, and they harbored point mutations in certain oncogenes not found in the human disease that are probably related to the chemical carcinogen (Ip C., 1996 J. Mammary Gland Biol Neopl 1: 37-47).
Other approaches involve the establishment of human cell lines. The most widely used human breast cancer cell lines are the MCF-7 (Soule H D. et al., 1973 J Natl Cancer Inst 51:1409-1416); the T-47-D (Engel L W. et al., 1978 Cancer Res. 38: 3352-3364); and the ZR-75-31 (Keydar I. et al., 1979 Cancer 51: 659-670). These lines express ER and progesterone receptors (PR) and are hormone responsive. In the last years, other cell lines have been established: PMC42 (Whitehead R H. et al., 1984 J National Cancer Institute 73:643-648); YMB-1 (Yamane M. et al., 1984 Hiroshima J Med Sci 33:715-720); VHB (Vandewalle B. et al., 1987 J Cancer Res Clin Oncol 113:550-558); IBEP-1, IBEP-2 and IBEP-3 (Siwek B. et al., 1998 Int J Cancer 76:677-683); BrCa-MZ-01 and BrCa-MZ-02 (Mobus V J. et al., 1998 Int J Cancer 77:415-423). A part of our knowledge on hormone regulation of cell growth has been deduced from experiments performed using these lines. Approximately twenty cell lines, which express hormone receptors, are available and have been used in other types of experiments. It is well-known that valuable initial information can be gathered from in vitro studies. However, cancer occurs in the context of a complex interaction with its surrounding environment, e.g., neighboring tissues, the immune system, hormones, and environmental factors. Thus, there is a trend to study either xenografts in immune-suppressed mice (Clarke R. 1996 Breast Cancer Res and Treat 39: 69-86) or transgenic or knock-out mice (Amundadottir L T. Et al. 1996 Breast Cancer Res and Treat 39:119-135). A problem with most cell lines is that they are not metastatic unless they are transfected with different growth factors, and also the high costs of immuno-suppressed mice. In mice there are few models which have been used to study hormone regulation in tumor growth, among than them MXT model and the GR mice. The former is a mammary tumor which is maintained by syngeneic transplantation and expresses high levels of ER and PR (Kiss R. et al. 1989 Cancer Res 49:2945-2951). The latter is a E) strain of mice which develops pregnancy-dependent tumors (Sluyser M. and Van Nie R. 1974 Cancer Res 34:3253-3257). No cell lines have yet been developed from mouse tumors which maintain steroid receptor expression except for a tumor induced in c-erbB2 transgenic mice (Sacco M G. Et al 1998 Breast Cancer Res and Treat 47:171-180).
The inventors have developed an experimental model in which ductal metastatic, progestin-dependent (PD) mammary adenocarcinomas are induced by the continuous administration of MPA to BALB/c female mice; these tumors express high levels of ER and PR (Lanari C. et al 1986 Cancer Letters 33: 215-223., Molinolo A A. et al 1987 JNCI 79:1341-1350), and are maintained through syngeneic serial passages in MPA-treated mice. By transplantation into untreated mice, the inventors have been able to generate progestin-independent (PI) tumor lines that retain the expression of ER and PR. In in vitro studies, using primary and secondary cultures of one of these PD tumor lines (C4-HD), the inventors were able to demonstrate that MPA stimulates directly cell proliferation and that antiprogestins inhibit cell growth even at very low concentrations (Dran G. et. al. 1995 Breast Cancer Res and Treat 35: 173-186).
Earlier in the characterization of the experimental model, with the aim of further dissecting certain aspects of the hormonal response, the inventors developed a technique to obtain purified epithelial or fibroblastic primary cultures from these tumors (Dran G. et al. 1995 Breast Cancer Res and Treat 35:173-186). Although primary cultures are an excellent tool to study the direct effect of hormones on cell proliferation, the approach is time consuming, e.g., many different controls need to be performed in parallel, to be able to bypass the inherent heterogeneity of every primary culture, and standardize the findings.
To overcome these shortcomings, in the setting of the characterization of the different isoforms of the PR expressed by the tumors, the inventors decided to attempt to develop continuous cell lines in which different parameters regarding hormone dependence could be studied. The inventors were able to develop four cell lines derived from one PD tumor and one from a PI tumor. This is the first description of mouse mammary adenocarcinoma cell lines obtained in non transgenic animals which express ER and PR.
The inventors provided a cell line panel for screening hormones and related molecules.
SUMMARY OF THE INVENTION
In accordance with the present invention, cell lines and methods for screening hormones are provided.
The present invention relates two mouse mammary adenocarcinoma cell lines MC4-L1 and MC4-L3, which are derived from a murine progestin-dependent CC4-HD tumor, wherein the cell lines express ER and PR and are deposited in the American Type Culture Collection (ATCC) as Accession number PTA-889 and PTA-891.
The present invention also relates a cell line, MC4-L2, obtained by subcloning of cell line MC4-L1 deposited with the ATCC as Accession number PTA-892
The present invention also relates to a mouse mammary adenocarcinoma cell line MC7-L1 which is derived from the murine progestin-independent C7-H1 tumor, wherein the cell line expresses ER and PR and have been depo
Lanari Claudia
Luthy Isabel
Molinolo Alfredo
Fulbright & Jaworski LLP
Helms Larry R.
Yu Misook
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