2012Unpublished venueRequires access

Inhibitory effects of RNA interference on expression of matrix metalloproteinase-2 gene and malignancy behaviours of ovarian cancer cells

Qingyuan Song, Xiujie Sheng, Yingqun Zhou, Zhen Li, Man Sun, Zhihui Wang

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Abstract

Objective To investigate the inhibitory effects of RNA interference (RNAi) on the expression of matrix metalloproteinase-2 (MMP-2) gene and growth, adhesion,invasiveness and migration of ovarian cancer cells. Methods One specific target sequence of MMP-2 gone and one non-specific sequence (NC group) were chosen,the medium DMEM as blank group.After transfection of ovarian cancer OVCAR-3 cells, the RT-PCR and Western blot were used to detect mRNA and protein expression of MMP-2 gene, the growth ability was detected by MTT assay, the abilities of adhesion was detected by cell adhesion assay, the invasion and migration were detected by Matrigel invasion assay and wound healing assay. Results By contrast to the NC group,the mRNA expression was decreased by 73.8 %,78.8 % and 78.4 %(P< 0.05) in 24 h,48 h and 72 h after transfection and protein expression was decreased by 72.6 %,81.2 % and 76.4 %(P< 0.05) respectively at the same time. The 48 h group had the most efficient inhibitory effect. Cell growth curve revealed that cell growth was not significantly inhibited (P> 0.05). Adhesion was significantly reduced,the inhibitory rate was 55.0 % at 60 min and 44.8 % at 90 min (P< 0.05),respectively. Invasion and migration were significantly reduced as well,the inhibitory rate on invasion and migration were 29.7 % and 35.8 %(P<0.05), respectively. Conclusion siRNA mediated MMP-2 down-regulation in ovarian OVCAR-3 cells can inhibits its adhesion,invasion and migration,but do not significantly affect its growth,suggesting a important target to ovarian cancer gene-therapies. Key words: Ovarian neoplasms;  RNA interference;  Matris metalloproteinase 2;  Neoplasm metastasis;  Gene therapy

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Objective To investigate the inhibitory effects of RNA interference (RNAi) on the expression of matrix metalloproteinase-2 (MMP-2) gene and growth, adhesion,invasiveness and migration of ovarian cancer cells. Methods One specific target sequence of MMP-2 gone and one non-specific sequence (NC group) were chosen,the medium DMEM as blank group.After transfection of ovarian cancer OVCAR-3 cells, the RT-PCR and Western blot were used to detect mRNA and protein expression of MMP-2 gene, the growth ability was detected by MTT assay, the abilities of adhesion was detected by cell adhesion assay, the invasion and migration were detected by Matrigel invasion assay and wound healing assay. Results By contrast to the NC group,the mRNA expression was decreased by 73.8 %,78.8 % and 78.4 %(P< 0.05) in 24 h,48 h and 72 h after transfection and protein expression was decreased by 72.6 %,81.2 % and 76.4 %(P< 0.05) respectively at the same time. The 48 h group had the most efficient inhibitory effect. Cell growth curve revealed that cell growth was not significantly inhibited (P> 0.05). Adhesion was significantly reduced,the inhibitory rate was 55.0 % at 60 min and 44.8 % at 90 min (P< 0.05),respectively. Invasion and migration were significantly reduced as well,the inhibitory rate on invasion and migration were 29.7 % and 35.8 %(P<0.05), respectively. Conclusion siRNA mediated MMP-2 down-regulation in ovarian OVCAR-3 cells can inhibits its adhesion,invasion and migration,but do not significantly affect its growth,suggesting a important target to ovarian cancer gene-therapies. Key words: Ovarian neoplasms;  RNA interference;  Matris metalloproteinase 2;  Neoplasm metastasis;  Gene therapy

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Available abstract

Objective To investigate the inhibitory effects of RNA interference (RNAi) on the expression of matrix metalloproteinase-2 (MMP-2) gene and growth, adhesion,invasiveness and migration of ovarian cancer cells. Methods One specific target sequence of MMP-2 gone and one non-specific sequence (NC group) were chosen,the medium DMEM as blank group.After transfection of ovarian cancer OVCAR-3 cells, the RT-PCR and Western blot were used to detect mRNA and protein expression of MMP-2 gene, the growth ability was detected by MTT assay, the abilities of adhesion was detected by cell adhesion assay, the invasion and migration were detected by Matrigel invasion assay and wound healing assay. Results By contrast to the NC group,the mRNA expression was decreased by 73.8 %,78.8 % and 78.4 %(P< 0.05) in 24 h,48 h and 72 h after transfection and protein expression was decreased by 72.6 %,81.2 % and 76.4 %(P< 0.05) respectively at the same time. The 48 h group had the most efficient inhibitory effect. Cell growth curve revealed that cell growth was not significantly inhibited (P> 0.05). Adhesion was significantly reduced,the inhibitory rate was 55.0 % at 60 min and 44.8 % at 90 min (P< 0.05),respectively. Invasion and migration were significantly reduced as well,the inhibitory rate on invasion and migration were 29.7 % and 35.8 %(P<0.05), respectively. Conclusion siRNA mediated MMP-2 down-regulation in ovarian OVCAR-3 cells can inhibits its adhesion,invasion and migration,but do not significantly affect its growth,suggesting a important target to ovarian cancer gene-therapies. Key words: Ovarian neoplasms;  RNA interference;  Matris metalloproteinase 2;  Neoplasm metastasis;  Gene therapy

Key concepts: Transfection, Matrigel, Molecular biology, Gentamicin protection assay, Cell adhesion, Matrix metalloproteinase, Biology, Messenger RNA

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