A Radiation‐induced Murine Ovarian Granulosa Cell Tumor Line: Introduction of v‐ras Gene Potentiates a High Metastatic Ability

Abstract

A non‐metastatic epithelial tumor cell line, OV3121, was established from ovarian granulosa cell tumor in B6C3F1 mouse irradiated with 60Co‐gamma rays. OV3121 cells showed an epithelial morphology and grew in monolayer with a population doubling time of 28–30 h. The production of estradiol and the expression of cytokeratin confirmed the epithelial origin of the line. No pulmonary metastasis was observed from solid tumors after subcutaneous (s.c.) injection or after intravenous (i.v.) injection of a clonal subline, OV3121–1 cells. We examined the experimental metastasis of individual clones of OV3121–1 cells, containing various introduced viral oncogenes: v‐Ha‐ras, v‐Ki‐ras, v‐fms, v‐mos, v‐raf, v‐src, v‐sis, v‐fos and v‐myc. Among them, only OV3121–1 cells with v‐Ha‐MuSV or v‐Ki‐MuSV produced lung colonies at high frequencies. In a more detailed analysis, the v‐Ha‐ras transfectants OV‐ras4 and OV‐ras7 were found to form colonies in various organs by metastasis from tumors after s.c. injection, as well as lung colonies after i.v. injection. Moderately metastatic OV‐ras7 cells showed high gelatinolytic activity at 72 kDa (MMP‐2) and 92 kDa (MMP‐9) as compared with the parental OV3121–1 and OV‐Neo control cells by zymographic analysis. However, more metastatic OV‐ras4 cells produced progressively weaker bands of 72 kDa gelatinolytic activity. No gross alterations in the expression of MMP‐1, MMP‐3, TIMP‐1 and TIMP‐2 transcripts were detected in these cell lines. These results suggest that this ovarian granulosa cell tumor line may provide a useful system for understanding the mechanisms by which oncogenes influence the occurrence of metastasis.

Comprehensive Analysis of Mouse Cancer/Testis Antigen Functions in Cancer Cells and Roles of TEKT5 in Cancer Cells and Testicular Germ Cells

“…We carried out KD of the remaining 84 genes by introducing two different siRNAs corresponding to each CTA gene into the cancer cell lines and estimating the changes in cell number. Those lines included melanoma (B16 [13] and B16C2W [14]), lung tumor (3LL [15] and KLN205 [16]), breast tumor (Ehrlich [17], MM46 [18], and FM3A [19]), liver tumor (Hepa1-6 [20] and MH134-TC [21]), bladder tumor (MBT-2 [22]), ovarian tumor (OV3121 [23] and OV2944-HM-1 [24]), and colon tumor (colon-26 [25]) cells. The combinations of genes and tested cell lines are shown in Table S4.…”

Minimum environmental enrichment is effective in activating antitumor immunity to transplanted tumor cells in mice

“…The tumor cell line OV3121, derived from an ovarian granulosa cell tumor from female B6C3F1 mice after irradiation with high dose-rate gamma rays [37], was obtained from Health Science Research Resources Bank (Osaka, Japan). OV3121 cells cultured in Roswell Park Memorial Institute 1640 medium supplemented with 10% fetal calf serum were trypsinized and suspended in normal saline to obtain 5 × 10 6 cells/ml cell suspensions.…”

The Ras family of GTPases in cancer cell invasion

“…The 92-kDa type IV collagenase/gelatinase B/matrix metalloproteinase-9 (MMP-9) is one MMP most frequently associated with Ras [85 -91]. Overexpression of the 72-kDa MMP-2 has also been linked to this oncogene in vitro [86] and in human malignancy [45]. Laminin induces the expression of MMP-2 through phospholipase D (PLD), an enzyme that cleaves phosphatidylcholine into PA and choline [92].…”