Effects of THY1 Gene on Growth of Epithelial Ovarian Cancer SKOV3 Cell Lines in Vitro and in Vivo

Abstract

Objective To construct THY1 gene eukaryotic expression plasmid and study its effects on epithelial ovarian cancer SKOV3 cell lines. Methods The gene fragment code for THY1 was obtained from human normal ovarian tissues using RT-PCR, and inserted into the eukaryotic expression plasmid pcDNA3.1(+ ) to construct the recombinant plasmid pcDNA3.1(+ )-THY1, which was transfected into SKOV3 cells from March 2005 to March 2007. Experimental cells were classified into 3 groups: SKOV3-THY1 group, SKOV3-Null group and SKOV3 group. The expression of THY1 mRNA and its protein were examined by RT-PCR and Western blot. The cell proliferation and cell cycles were evaluated by methyl thiazolyl tetrazolium (MTT)assay and flow cytometry. Tumor sizes were measured in vivo tumorigenicity studies. Results The gene fragment of THY1 was correctly inserted into the eukaryotic expression plasmid pcDNA3.1(+ ) as verified by PCR, restriction endonucleases digestion and DNA sequencing, and the pcDNA3.1(+ )-THY1 has been transfected into SKOV3 cells and obtained stable expression by RT-PCR and Western blot methods. The cell inhibitory rate of the SKOV3-THY1 group(56.6% at the 5th day) was higher than the SKOV3-Null group(12.5%), there was significant difference between them(P 0.05). In vivo tumorigenicity studies, tumors grew quite rapidly in the null transfectants compared with the THY1-transfected cells over the next 4 weeks. Conclusion THY1 transfection can inhibit the growth of epithelial ovarian cancer SKOV3 cells in vitro and in vivo. THY1 gene may play an important role in generation and development of ovarian cancers. Key words: gene; THY1; eukaryotic expression plasmid; transfection; ovarian neoplasm