Protein profilling and investigation on l myc c myc and fgf 3 genes in patients with ovarian malignancy vs non malignant individuals with normal ovaries

Abstract

newlineThis study aims at expression of L-MYC, C-MYC and FGF-3 genes in ovarian carcinoma patients conducted using blood samples from 50 ovarian cancer patients and 50 non-malignant females. PCR amplification of exon-1 of L-MYC gene and exon-3 of C-MYC gene were classified as low or highly expressing on the basis of their intensities after electrophoresis. The expression of 165 bp segment of exon-3 of C-MYC gene in cases was low as compared to controls indicating that expression of C-MYC gene is involved in the progression of ovarian cancer. Whereas, the expression of 198 bp segment of exon-1 of L-MYC gene in cases and controls was same. The segment of exon-3 of FGF-3 gene was not amplified in the present study. DNA sequencing of 198 bp segment of L-MYC gene and 165 bp segment of C-MYC gene showed 179 and 185 nucleotide changes in the exon-1 and exon-3 region of L-MYC and C-MYC genes, respectively. The exon-1 and exon-3 regions of L-MYC and C-MYC genes showed 87 and 98 nucleotide changes in controls also. The changes observed in the present study in nucleotide sequences have not been reported as yet. One dimensional gel electrophoresis of 3 selected cases and a control showed distinctive changes in six specific protein fractions of the molecular weight 250 kDa, 150 kDa, 75 kDa, 25 kDa, 10 kDa and lt 10 kDa between cases and control. Two dimensional gel electrophoresis of cases and control showed differential protein fractions. 46 protein spots were visible in control and 56 fractions were found in cases. One distinct protein at 52 kDa was found to be highly expressive in a highly metastatic, non-operated and non-treated stage IV case and less expressive in other two cases and a control. Mass spectrometry of ~52 kDa protein had 80% sequence coverage with chain A of crystal structure of 351 amino acid sequence of alpha-1-antitrypsin. The amino acid sequence of this protein showed changes from positions 39-101 and 244-259 with deamidation and oxidation.