Wen-xin Liu1, Xi-shan Hao. 1. Department of Pelvic Oncology, Cancer Hospital & Institute, Tianjin Medical University, Tianjin 300060, China.
Abstract
OBJECTIVE: To screen, clone and identify the cDNA fragments of human endometrial carcinoma-related genes,and explore the molecular mechanism of endometrial carcinogenesis. METHODS: Pure endometrial glandular epithelial cells and endometrial carcinoma cells were obtained by laser capture microdissection (LCM). RNA from these cells was isolated, and differentially expressed gene fragments that were specialy relevant to endometrial carcingenesis were identified by using fluorescence differential display reverse transcription polymerase chain reaction (FDD-PCR). The selected fragments were cloned, sequenced and verified by reverse Northern blot analysis, and positive fragments were BLAST analysed and compared with those in Genbank. RESULTS: 38 differential fragments were isolated, 3 of which were expressed more abundantly in normal endometrium and 35 were highly expressed in endometrial carcinoma. 10 fragments were recoverd, cloned and sequenced, confirmed by reverse Northern blot analysis, among which 6 fragments were positive. BLAST analysis showed that T1.1 was homologous to cyclin-dependent protein kinase 7 (CDK7, 99%); L1.9 was homologous to protein phosphatase 1 regulatory (inhibitor) subunit 12A (PPP1R12A, 99%); L1.21 and L1.22 were homologous to cellular repressor of E1A-stimulated genes 1 (CREG, 100%); L1.25 and L1.26 were homologous to solute carrier family 39 (zinc transporter) member 10 (SLC39A10, >98%). CONCLUSION: Gene fragments related to endometrial carcinoma have been obtained by applying LCM and FDD-PCR. To our knowledge it is the first time that the correlation between CDK7, PPP1R12A, CREG, SLC39A10 and endometrial carcinoma is discovered at mRNA level, and their role in molecular mechanism of cancinogenesis is discussed. CDK7, CREG, SLC39A10 as new candidate oncogene and PPP1R12A as new candidate anti-oncogene are worthy of being further investigated in the future.
OBJECTIVE: To screen, clone and identify the cDNA fragments of humanendometrial carcinoma-related genes,and explore the molecular mechanism of endometrial carcinogenesis. METHODS: Pure endometrial glandular epithelial cells and endometrial carcinoma cells were obtained by laser capture microdissection (LCM). RNA from these cells was isolated, and differentially expressed gene fragments that were specialy relevant to endometrial carcingenesis were identified by using fluorescence differential display reverse transcription polymerase chain reaction (FDD-PCR). The selected fragments were cloned, sequenced and verified by reverse Northern blot analysis, and positive fragments were BLAST analysed and compared with those in Genbank. RESULTS: 38 differential fragments were isolated, 3 of which were expressed more abundantly in normal endometrium and 35 were highly expressed in endometrial carcinoma. 10 fragments were recoverd, cloned and sequenced, confirmed by reverse Northern blot analysis, among which 6 fragments were positive. BLAST analysis showed that T1.1 was homologous to cyclin-dependent protein kinase 7 (CDK7, 99%); L1.9 was homologous to protein phosphatase 1 regulatory (inhibitor) subunit 12A (PPP1R12A, 99%); L1.21 and L1.22 were homologous to cellular repressor of E1A-stimulated genes 1 (CREG, 100%); L1.25 and L1.26 were homologous to solute carrier family 39 (zinc transporter) member 10 (SLC39A10, >98%). CONCLUSION: Gene fragments related to endometrial carcinoma have been obtained by applying LCM and FDD-PCR. To our knowledge it is the first time that the correlation between CDK7, PPP1R12A, CREG, SLC39A10 and endometrial carcinoma is discovered at mRNA level, and their role in molecular mechanism of cancinogenesis is discussed. CDK7, CREG, SLC39A10 as new candidate oncogene and PPP1R12A as new candidate anti-oncogene are worthy of being further investigated in the future.