LNCaP Cells: Transfection and Cell Culture Protocols
The LNCaP cell line (Lymph Node Carcinoma of the Prostate) is derived from human prostate adenocarcinoma cells from a lymph node metastasis. LNCaP cells are androgen-sensitive adherent epithelial cells that grow in aggregates as well as in single cells. LNCaP cells grow well in vitro and also may be injected into athymic nude mice for cell line derived xenograft (CDX) purposes (laboratory xenograft contract research services are provided by Altogen Labs – see Xenograft Services). These cells also develop tumors at the injection site and have a similar in vivo doubling time. LNCaP cells have an aneuploid chromosome number (76-91) and express Prostate Specific Antigen (PSA).
LNCaP-specific transfection reagent is commercially available from Altogen Biosystems.
Origin of the LNCaP Cell Line
The LNCaP cell line was established in 1977 by J.S. Horoszewicz, et al., from a needle aspiration biopsy of a metastatic lesion in the left supraclavicular lymph node of a 50-year-old Caucasian male with confirmed prostate carcinoma.
Growing LNCaP in vitro and in vivo (xenograft)
LNCaP cells are epithelial cells with adherent (and semi-adherent) qualities when growing in cell culture. The LNCaP cell line is sensitive to the hormones estrogen and androgen, which can be used to modulate growth. LNCaP cells are androgen dependent and resistant to human fibroblast interferon when grown in culture.
In vivo, tumor development is sex specific. Studies have shown male mice injected with LNCaP develop adenocarcinoma earlier and with greater frequency than females. This likely has to do with hormonal differences between the sexes and indeed, tumor development frequency is tightly correlated with androgen levels regardless of sex. Interestingly, once a tumor is established, growth is unrelated to gender or hormonal characteristics of the cancerous mouse.
LNCaP Xenograft
In culture, human prostate cells proliferate at an acceptable rate and number of passages. However, unlike other cancers, prostate cancer xenograft models are lacking due to the inability of immunocompetent mice to be a well-suited environment to grow human prostate tumors. This includes the lack of malignant prostate cell lines that are needed for this highly malignant cancer. Prostate cancer cell lines that are likely candidates for xenograft studies end up losing expression of key proteins (i.e. hPAP, PSA).
Multiple groups have reported on the genetic variation for which LNCaP cells are capable. Typically, even with additional genetic variances observed, cell sublines can be traced back to parental profiles. However, upon analysis, LNCaP cells inoculated in castrated mice underwent dramatic genetic and morphological changes. Further cytogenetic analysis exhibited a loss of the Y chromosome, along with major chromosomal reorganization. In a response to major tumor cell infiltration, stromal cells in the tumor microenvironment may become activated and growth factors at an elevated level. Co-culture experiments to better understand the in vivo interaction of LNCaP and stromal cells.
Characteristics of LNCaP cells
Highly sensitive androgen receptors are present in the cytosol of LNCaP cells both in culture and in tumors, making LNCaP a highly androgen-sensitive cell line. Cells grown in culture and in prostate tissue both secrete phosphatase. The cells have cytosolic estrogen receptors, with cell growth and acid phosphatase production responding to hormonal modulation (the cells are sensitive to 5-alpha-dihydrotestosterone).
LNCaP cells have an hypotetraploid human male karyotype with a modal number 76 to 91 and several marker chromosomes. Most cells have a chromosomes number of 84 (22%), but higher and lower numbers are seen in a lower frequency of cells, with modal number 86 in 20% of cells and 87 in 18% of cells. The rate of cells with ploidies higher than this drops dramatically to roughly 6%. The cell line is fairly stable, and more importantly, the malignant qualities of the cells are well maintained, making LNCaP useful for oncology research, specifically the study of prostatic adenocarcinoma for which they have been widely used.
Functional differentiation is also maintained, and is evidenced by the expression of Prostate Specific Antigen (PSA), and Human Prostatic Acid Phosphatase (hPAP). These are the two major prostate epithelium-specific differentiation antigens which serve to indicate the presence of active cytosolic androgen receptors in prostate cells. LNCaP is the only commercially available cell line that expresses hPAP, making it the only line available to study transcriptional regulation in the hPAP gene. Other prostate cancer cells lines, such as PC-3, are unable to express hPAP for unknown reasons. Due to their sensitivity to androgen and being functionally differentiated, LNCaP is a good model for studying transcriptional regulation in genes of the prostate. This is a result of many gene products in prostate cells being prostate-specific as a result of androgen-dependent processes.
Links
Transcriptional regulation of the human prostatic acid phosphatase gene
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