Isolation and characterization of two novel colon cancer cell lines from Chinese patients.

Incidence of colon cancer has increased rapidly in China. Although many colon cancer cell lines have been established previously, most of them were derived from patients from western countries. Epidemiological, clinical, cytogenetic, and molecular biological studies showed that there are considerable differences between Chinese and western countries colon cancer patients. Therefore, establishment of novel colon cancer cell line from Chinese is useful for studying the racial difference of this disease and can be important for studying the pathogenesis of colon cancer in China. In our laboratory, two novel continuous human colon cancer cell lines, SHT-1 and SHH-1, have been established in vitro from Chinese patients, and both cell lines have been passaged for 4 yr, and they have been continuously subcultured with more than 800 population doubling and without signs of senescence. Both cell lines were obtained from primary tumor tissues during colon cancer surgery. Cells grew rapidly with a doubling time of 36-39 h and a plating efficiency of 26-28%. These cells exhibited an epithelial morphology and expressed cytokeratin. Tumor developed in severe combined immunodeficient (SCID) mice 4-6 wk after inoculated subcutaneously with the cultured cancer cells. Karyotypic analysis and comparative genomic hybridization (CGH) analysis in SHT-1 cells revealed a hypertriploid modal number of 76 with numerous numerical and structural abnormalities previously linked to colon cancer. In another cell line (SHH-1), CGH analysis revealed that -1p13 was the only cytogenetic anomaly.

Uveal melanocytes produce matrix metalloproteinases-2 and -9 in vitro.

The purpose of the present study was to investigate the expression of matrix metalloproteinase (MMP)-2 and MMP-9 by cultured human uveal melanocytes, and to test the effects of 12-O-tetradecanoyl-phorbol-13-acetate on the expression of these MMPs. Gelatin zymography of conditioned culture medium from four cultures of human uveal melanocytes (two cultures of iridal melanocytes and two cultures of choroidal melanocytes) detected MMP-2 (72 kDa) and a relatively small amount of MMP-9 (92 kDa), both in the latent form. RT-PCR analysis revealed the MMP-2 mRNA and MMP-9 mRNA in cultured uveal melanocytes. Addition of 12-O-tetradecanoyl-phorbol-13-acetate (10 ng/ml) to the culture medium caused an increase of production of MMP-2 and MMP-9 by cultured uveal melanocytes, and also stimulated the transcription of MMP-2 and MMP-9 of these cells.