1991 Fiscal Year Final Research Report Summary
Analysis of the progression mechanism of human hepatocellular
| Project/Area Number |
02670572
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Digestive surgery
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| Research Institution | Yamanashi Medical College |
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Principal Investigator |
NAGAHORI Kaoru Yamanashi Medical College, 医学部, 助手 (00137035)
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| Co-Investigator(Kenkyū-buntansha) |
MIURA Kazuo Yamanashi Medical College, 医学部, 助手 (00221599)
YAMAMOTO Masayuki Yamanashi Medical College, 医学部, 講師 (30158307)
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| Project Period (FY) |
1990 – 1991
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| Keywords | Hepatocellular carcinoma / mito chondrial gene / nm23 gene / topoisomerase gene / RT-PcR / RTーPCR法 / トポイソメレ-ス遺伝子 |
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| Research Abstract |
We analyzed the changes of topoisomerase gene, mitochondrial gene and anti-metastatic gene nm 23 in human HCC and noncancerous tissues. <Specimens> Both malignant and noncancer ous tissues of HCC patients were used. <Kethods> Oligonucleotides homologous to the genes were synthesized using the DNA synthesizer. They were used as probes for the hybridization and as primers for RT-PCR reaction. Total cellular DNA was isolated by phenol/chloroform extraction and total cellular RNA was isolated by density centrifugation in a cecium chloride gradient. Amplification of topoisomerase gene and nm 23 gene were assayed using Southern blot hybridization technique. The region coding cytochrome C oxidase in mitochondrial DNA were studied by RFLP after PCR amplification. Expression of nm 23 MRNA was semiquantitated by primeri-directed reverse transcription (RT) and subsequent polymerase chain reaction (PCR) with densitometry. beta-actin mRNA was used as the internal control. <Results> Amplification of topoisomerase gene and nm 23 gene were not shown. The difference of restriction fragments in mitochond-rial DNA between cancerous and noncancerous portion was not detected. Expression of nm23 mRNA was shown as relative density unit (RDU) following normalization to beta-actin expression. RDU of normal liver, liver cirrhosis and HCC were 1.23<plus-minus>0.35 (n=4), 0.78<plus-minus>0.44 (n=6) and 1.38<plus-minus>0.59 (n=7), respectively. RDU of HCC from patients with intrahepatic metastasis were lower than that of HCC without intrahepatic metastasis (not significant). <Discussion> Despite the lack of knowledge about the true role of nm 23/NDP kinase, it might be useful to have advanced information regarding the prognosis of patients sufering fro
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