| Objective: The purpose of this study was to investigatetelomere length, hTRT protein, DNA content and ALT path inorder to explore the role of them in carcinogenesis ofesophageal epithelium.Methods: Telemere length, hTRT content and DNAcontent were analyzed quantitatively by flow cytometry (FCM)combined fluorescent in situ hybridization (FISH),immunofluorescence staining (IFS) and DNA fluorescencestaining in 100 esophageal epithelium cells samples, including18 cases of normal esophageal epithelial cells, 16 cases of milddysplasia, 35 cases of severe dysplasia and 31 cases ofcarcinoma. Immunohistochemistry S-P method was used todetect the expression of hTRT and PML protein in cases of 44formalin-fixed, paraffine-embeded esophageal carcinomasamples. 11 cases of incisal margin normal esophageal tissueand 12 cases of para-tumorous esophageal epithelium dysplasiawere study as control.Results: 1.Value of Q-FISH in normal esophagealepithelium group, mild dysplasia group, severe dysplasia groupand cancinoma group was 50.83±8.86, 49.52±3.16, 36.96±8.02and 27.81±6.59 respectively. Value of Q-FISH in cancinomagroup was higher than that in severe dysplasia group (p<0.01),severe dysplasia group was higher than mild dysplasia group(p<0.01). There was a negative correlation betwen telomerelength and cytologic grade(r=-0.79, P<0.01). According to thestandard: telomere shorting was less than 80% of telomerelength of normal gorup, from esophageal percancerous cells tocarcinomous cells tlomere shorting rate was 56.25%(9/16),62.86%(22/35) and 83.87%(26/31) and showed increasedtendency (P>0.05). 2 FI value of hTRT in normal esophagealepithelium group, mild dysplasia group, severe dysplasia groupand cancinoma group was 0.95±0.14, 1.15±0.22, 1.47±0.22 and1.75±0.19. FI value of hTRT in cancinoma group was higherthan that in severe dysplasia group (p<0.01), severe dysplasiagroup was higher than mild dysplasia group (p<0.01). FI valueof hTRT was positive related with cytologic grade(r=0.84,P<0.01); According to the standard: value of FI>1.0 waspositive, expreesing rate of hTRT portain was 68.75%,91.43%and 96.77% in groups of mild dysplasia, severe dyaplsia andcarcinoma. Expressing rate in cancinoma group and severedysplasia group were higher than that in mild dysplasia group(p<0.01). From normal group to cancinoma group value of DI(DNA Index) was 1.01±0.32, 1.04±0.21, 1.10±0.28 and 1.51±0.36. DI value in cancinoma group was higher than that insevere dysplasia group (p<0.01), severe dysplasia group washigher than mild dysplasia group (p<0.01). There was positivecorrelation betwen value of DI and cytologic grade(r=0.60,P<0.01). With the cytologic grade progressing value of PI(proliferation index) increased, value of PI (29.46±5.50) incancinoma group was higher than that (19.04±5.39) in severedysplasia group. Value of PI was positive related with cytologicgrade (r=0.84, P<0.01). 4. There was show significant positivecorrelation betwen value of DI and PI(r=0.93, P<0.01), andnegative correlation between Q-FISH value of telomere and FIvalue of hTRT(r=-7.34, P<0.01), as well as Q-FISH value oftelomere vs. value of DI(r=-7.34, P<0.01). 5. The expression ofhTRT located in nucleus of esophageal epithelial cells andcancerous cells by Immunohistochemistry staining. The positiverate of hTRT in incisal margin normal tissue was zero, and thosein para-tumorous dysplasia tissues and cancinoma were41.67%(5/12) and 84.09%(37/44), there was instinct differenceof positive expression among them (P<0.01). Expression ofhTRT had no relationgship observed with age, gender,differentation grade, lymph node matastasis and invasive degreeof patients (P>0.05). 6. The expression of PML located innucleus by Immunohistochemistry staining. There was no caseshowing expression of PML in incisal margin normal tissue, andcases in para-tumorous dysplasia tissue and cancinoma were1(8.33%) and 5(11.36%) respectively. With the pathologicaldeveloping, positive case of PML seemed increasing, whereasthere was no instinct difference among them (P>0.05).Conclusion: In the process of pathological developing,telomere length decreased and hTRT, DNA content and PI value...
|
PDF Full Text Request