| Abstract |
Continuously growing lines of cancer cells have been invaluable for
studies on various types of cancer. For prostate cancer, only a few
different lines are available. We have derived three new prostate cancer
cell lines from tissues obtained at the time of prostatectomy. The tissues
were cut into small pieces with sterile scissors placed in keratinocyte
serum free medium (KSFM), centrifuged, resuspended in KSFM and layered
onto tissue culture plastic pre-coated with FCS. Growth was achieved by
culturing the disrupted tissue in KSFM supplemented with huEGF and 2% FCS.
When cell proliferation was evident, the cells were exposed to a
replication-defective retrovirus transferring the transforming HPV16 or
HPV18 E6 and E7 genes, as well as neoR, conferring Geneticin-resistance.
Transformed cells were then selected using Geneticin. All primary
cultures were maintained through crisis until a viable permanent cell line
had been established. These lines are termed OPCT-1, OPCT-2 and OPCT-3. In
a similar way, three new immortalized normal prostate epithelial cell
lines have been derived from prostatectomy tissue. These lines are OPCN-1,
OPCN-2 and OPCN-3. The doubling times (Td), calculated from a series of
growth curves, suggest that the tumor lines proliferate more rapidly than
the normal lines with Td of 1.1 +- 0.02 (SEM) days for OPCT-1, 1.8 +- 0.2
days for OPCT-2 and 1.9 +- 0.3 days for OPCT-3 compared to 2.7 +- 0.5 of
OPCN-1, 2.9 +- 0.5 for OPCN-2 and 2.5 +- 0.5 days for OPCN-3.
Immunohistochemistry results indicate that OPCT-1 is positive for
cytokeratins 18 (CK18+), 5 and 8 (CK5/8+), vimentin, MUC-1 and flotillin.
It is negative for desmin, alpha smooth muscle actin, and factor 8. The
other lines have similar profiles for these markers, although OPCT-2 and
OPCN-1 are only weakly positive for flotillin and OPCN-2 appears to be
negative. For MUC-1, OPCN-2 and OPCT-2 are weekly positive. All lines
appear to be negative for androgen receptor and positive for prostate stem
cell antigen (PSCA). OPCT-2 is negative for PSA whereas the other three
lines appear to be weakly positive. OPCT-3 and OPCN-3 have not been
examined yet. These lines should be valuable tools for prostate cancer
research.
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