Sunil K. Chatterjee

Elevated Expression of Ferritin H-Chain mRNA in Metastatic Ovarian Tumor

To identify genes associated with tumor metastasis, we prepared 5 cDNA libraries using mRNA from normal ovaries, paired primary and metastatic ovarian tumors, as well as paired cultured ovarian tumor cells. By differential screening, we identified 12 clones, which can be divided into 3 classes based on hybridization to various probes. Class 1 clones showed no reaction with the normal probe, slight or no reaction with the primary probe, but high reaction with the metastatic probe. Class 2 clones showed some reaction with normal and primary probes, but showed stronger reaction with the metastatic probe. Class 3 clones showed strong hybridization to the normal probe, slight or no reaction with the primary probe, and did not hybridize with the metastatic clone. These clones were further analyzed by determination of DNA sequence. One of the class 1 clones (clone 1) was identified as ferritin heavy chain. Northern blot analysis showed higher expression of ferritin H-chain in metastatic samples compared to primary tumor in 16/23 pairs of samples analyzed so far.

Analysis of the sequences of human β-1,4-galactosyltransferase cDNA clones

UDP-galactose:beta-1,4 N-acetyl glucosamine galactosyltransferase (4 beta GT) is a promising tumor marker for ovarian cancer. To study the role of 4 beta GT in malignant transformation at the molecular level human 4 beta GT cDNA and genomic clones were isolated and analyzed. For the isolation of 4 beta GT cDNA and genomic clones, a human fetal liver cDNA library in lambda gt11 and a human genomic library in EMBL-3B vectors respectively were screened using a 4 beta GT cDNA insert as the probe. Complete sequence of the cDNA clones were determined by subcloning in plasmid vectors, and compared with the published sequence of human liver 4 beta GT. Presence of various 4 beta GT exons in the genomic clones were determined by Southern blot analysis using specific oligodeoxynucleotide probes. Among the 5 cDNA clones isolated, 2 clones GTN 6 and GTN 17 were sibling clones and had a nucleotide sequence identical to the published 4 beta GT cDNA sequence, except at the 3-end, where these clones had 7 unique nucleotide sequences. One cDNA clone, GTN2 also had a nucleotide sequence identical to that of 4 beta GT, except for 3 G residues at the 5-end. One cDNA clone, GTN 1, had a unique sequence at the 5-end comprising of 74 nucleotides. Another clone, GTN 20, was unrelated to 4 beta GT. Analysis of genomic clones showed that 4 beta GT exons 3, 4, 5 and 6 were present in a 14 kb genomic clone, EMGT-4. Exon 1 was present in a separate 16 kb clone, EMGT-6.(ABSTRACT TRUNCATED AT 250 WORDS)