| PrefaceHepatocellular carcinoma ( HCC) is a very common malignancy in Asian countries and in South Africa, the prognosis of which remains poor despite the development of improved diagnostic modalities and advancement in treatment. With the development of molecular biology techniques, telomeres are found, which are specialized structures at the terminal regions of eukaryotic chromosomes , and are considered to be important in protecting and stabilizing the chromosomal ends. Normal human somatic cells correspondingly have a limited pro-liferative capacity. Telomerase is expressed in germ line cells and in immortal cells, but not in most somatic cells. Since a very high level of telomerase activity has been found in a high percentage of various cancer cells. Generally, malignant tumors have strong telomerase activity, while the activity in surrounding non -cancerous lesions is weak or below detectable level. Thus ,reactivation of telomerase represents an important step toward unlimited proliferation and indefinite survival of cancer cells. Human telomerase reverse transcriptase (hTERT) is a human telomerase catalytic subunit . A significant correlation between hTERT expression and telomerase activity has been reported . Presently, hTERTmRNA was detected by RT - PCR analysis, and hTERT immunohistochemistry are also applicable for evaluation as surrogate markers of telomerase expression at the cellular level and in archival paraffin embedded specimens, we applied to the method of situ hybridization to detect hTERTmRNA expression at the cellularlevel and in archival paraffin embedded tissue in Human Hepatocellular Carcinoma and Hepatocirrhosis in order to study the effect of hTERT during the transformation hepatocirrhosis towards the hepatocellular.Materials1. Sample collectionSixty - nine patients were enrolled in this study. The pathological diagnosis of twenty - eight patients was HCC, and that of forty - one was chronic hepatocirrhosis. The pathological diagnosis is decided by three pathological doctors. All tissues samples were obtained from the pathological department of the first hospital medical university of china. They are paraffin - embedded samples during 2002-2003 years.2. ReagentsDEPC hTERTmRNA situ hybridization kit poly - L - LYSINE DAB 2 SSC 0.2 SSC 0.5 SSCMethods1. Situ hybridizationAccording to the introduces of hTERTmRNA situ hybridization kit, reagents are one by one applied to 4um - thick paraffin - embedded sections.2. AnalysisData were expressed as means SD, and were statistically analyzed by the x~2 - test and F - test . A level of p <0. 05 was accepted as statistical significance.Results22 of 28 HCC were hTERTmRNA positive , positive rate is 78.6% .average grey value is 192. 179 ± 5. 97, and 7of 41 hepatocellular cirrcosis were hTERTmRNA positive , positive rate is 17. 1% , average grey value is 194. 325 ±3.07(p<0.01).DiscussionHepatocellular carcinoma ( HCC) is a very common malignancy, the prognosis of which remains poor despite the development of improved diagnostic modalities and advancement in treatment. With the development of molecular biology techniques, telomeres are found ,which are specialized structures at the terminal regions of eukaryotic chromosomes and are considered to be important in protecting and stabilizing the chromosomal ends. Human telomerase reverse transcriptase (hTERT) is a human telomerase catalytic subunit. A significant correlation between hTERT expression and telomerase activity has been reported. We applied to the method of situ hybridizaion to detect hTERTmRNA expression at the cellular level and archival paraffin embedded tissue in human hepatocellular carcinoma and hepatocirrhosis.The result here indicated that hTERTmRNA was expressed more frequently in HCC tissues than in hepatocellular cirrcosis tissues and the intensities of hTERTmRNA expression was stronger in tumor tissues than in hepatocellular cirrcosis. So reactivation of telomerase represents an important step toward unlimited proliferation and indefinite survival of cancer cells.In remained unclear ,however, whether the increases in the level of telomerase activity with degree of histological differentiation of HCC correlates with the increase in telomerase activity in individual carcinoma cells or with the increase in frequency of telomerase positive carcinoma cells. In the present study, we found that the increase in frequency of hTERTmRNA positive cells plays a key role in the transformation of hepatocirrhosis towards the hepatocellular.We also study the relation between the levels of hTERTmRNA in the HCC tissues and clinical pathological parameter . The result suggests that the correlation between the signal intensity and degree of histological differentiation was obvious. In well -differentiated HCC nodules ,the grey values is 192. 34 ±7.14, in the moderately and poorly differentiated HCC, the grey values respectively is 190. 94 ± 5. 05,184. 07 ± 6. 87. Although the intensity of immunostaining in moderately or poorly differentiated . HCC was often higher than that in well - dif-... |
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