ABSTRACT
<p><b>OBJECTIVE</b>To investigate the inhibitory effects of an antisense PC cell derived growth factor (PCDGF) vector on proliferation and invasion of highly malignant ovarian cancer cell lines Sw626 and A2780 cells, and preliminarily explore the related mechanisms.</p><p><b>METHODS</b>MTT assay and Boyden chamber in vitro invasion assay were employed to detect the changes of proliferation and invasion ability in the Sw626 and A2780 cells transfected with anti-sense PCDGF. The expression levels of cyclin D1 and CDK4 proteins before and after transfection were detected by Western blotting. The effects on the expression and activity of MMP-2 were evaluated by quantitative RT-PCR and zymography, respectively.</p><p><b>RESULTS</b>Comparing with the blank group, the proliferation inhibition rate of the Sw626 and A2780 cells transfected with anti-sense PCDGF was 72.9% and 70.9%, respectively, and the invasion ability was inhibited by 62.9% and 59.0%, respectively. The levels of cyclin D1 and CDK4 protein expression in antisense PCDGF transfected cells were 0.38 +/- 0.08 and 0.37 +/- 0.13, respectively, all significantly lower than 0.84 +/- 0.11 and 0.64 +/- 0.11, respectively, in the blank group (P < 0.01). The MMP-2 mRNA expression level in antisense PCDGF transfected cell group was 0.66 +/- 0.11, not significantly decreased in comparison with 0.89 +/- 0.09 in the blank group (P > 0.05), but the activity of MMP-2 was inhibited significantly.</p><p><b>CONCLUSION</b>The antisense PCDGF vector may inhibit markedly the proliferation and invasion of highly malignant ovarian cancer cells, and partially reverses their malignant phenotype. It seems to be related with down-regulating the expression of cyclin D1 and CDK4 and inhibiting the activity of MMP-2. Our findings indicate that PCDGF may become a new target for antisense gene therapy of ovarian cancer.</p>
Subject(s)
Female , Humans , Cell Adhesion , Cell Line, Tumor , Cell Proliferation , Cyclin D1 , Metabolism , Cyclin-Dependent Kinase 4 , Metabolism , DNA, Antisense , Down-Regulation , Genetic Vectors , Intercellular Signaling Peptides and Proteins , Genetics , Metabolism , Matrix Metalloproteinase 2 , Genetics , Metabolism , Neoplasm Invasiveness , Ovarian Neoplasms , Metabolism , Pathology , RNA, Messenger , Metabolism , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To investigate the frequency of microsatellite instability (MSI) and loss of heterozygosity (LOH) at human leucocyte antigen(HLA) class I loci in cervical carcinoma and the detailed deletion mapping in this region.</p><p><b>METHODS</b>LOH and MSI of HLA class I genes were analyzed in 30 paired blood and tumor samples by PCR based single-stranded length polymorphism (PCR-SSLP).</p><p><b>RESULTS</b>Of the 30 cases, 23(76.7%) showed LOH at one or more loci. Higher frequencies of LOH were found at four loci: C3_2_11 (50%), C1_4_4 (37%), C1_2_5 (36.7%), D6S276 (48.3%). MSI was found in 20 out of 30 cases (66.7%).</p><p><b>CONCLUSION</b>The data suggest that the LOH and MSI of HLA class I gene might participate in the carcinogenesis of cervical carcinoma. Meanwhile, the minimal deletion region might be defined between C1_2_5 and C3_2_11, thus providing the evidence for cloning the tumor suppressor genes associated with cervical carcinoma.</p>