| Objective To establish a human ovarian carcinoma cells with directional highly lymphatic metastasis and study of their biological characteristics and of differential expression related gene of metastasis.Methods The clone cells of ovarian carcinoma, SKOV3,were inoculated into the hind foot pad of nude mice.The cancer cells of lymph node metastasis foci were transplanted into nude mice again when the metastasis nude mice of were observed.After repetition of this procedure for 3 cycles,the metastasis rate and the metastasis paths were observed in nude mice of every passage. Using limited dilution method to separate and select colonial cells with directional highly lymphatic metastasis potentials from the lymphatic metastasis of human ovarian carcinoma cell line SKOV3.The cells with biological characteristics were assayed by series of growth curve,HE staining,cerotype analysis,nude mice transplantation and immunohistochemistry.Utilizing gene array technology to study the differential expression related gene of metastasis among every generation cells.Results Established a series of cell lines from lymph node metastasis and designated them as SKOV3-PM1,SKOV3-PM2 and SKOV3-PM3 cell strain.When the cells were injected into the hind footpad of nude mice producing 100%(10/10) spontaneous Lymphatic metastasis.The lymphatic metastasis rates were stable.The metastasis paths were single;the proliferation ability was increased in cancer ceils of every passage in vivo and vitro after passaging cancer cells of lymphatic metastatic loci for 3 cycles in nude mice.Cytogenesis study showed the karyotins of SKOV3-PM3 had mode ranged from 83 to 89,and the SKOV3 from 91 to 105.The former grows more quickly than the latter,their cell population doubling time being 22.7 hours and 49.9 hours, respectively.Immnocytochemical study demonstrated all cell lines were still maintained polygonal epithelial cells,exhibiting features of human carcinoma.Flow cytometry revealed the proportion of cells in DNA synthesis and mitosis was higher in SKOV3-PM than in SKOV3.In this study,there are 60 genes difference expression among 90 genes related to metastasis.To compare SKOV3-PM2,SKOV3-PM3 with SKOV3,there are 31 genes up-regulated,1 gene down-regulated at the same time.Conclusions The ovarian carcinoma sublimes with directional highly lymphatic metastasis potential have been established.The cells could be provided a good experimental material for further investigation the possibilities mechanism of metastasis and invasion ability of ovarian carcinoma.Screening out different expression genes related to directional highly lymphatic metastasis. |
