Title PCNA immnolabelling in colorectal c. Citation Acta Medica Nagasakiensia. 1993, 38

Transcription

1 NAOSITE: Nagasaki University's Ac Title Author(s) A Study sequential demonstra immnolabellg c Yoshida, Kazuya; Tomita, Masao itation Acta Medica Nagasakiensia. 1993, 38 Issue Date URL Right This document is downloaded

2 Acta Med. Nagasaki 38:15-20 A Study sequential demonstration nucleolar organizer regions immnolabellg carcomas Kazuya Yoshida (Director: Pr. Masao Tomita) The First Department Surgery, Nagasaki University School Medice, 7-1 Sakamoto 1-chome, Nagasaki 852, Japan Summary: The possibility a sequential demonstration nucleolar organizer regions (NORs) proliferatg cell nuclear antigen () by argyrophil () method immunostag, respectively, was explored carcomas. Both argyrophilic protes NORs (s) were clearly visualized when immunostag by labelled streptavide-biot (LSAB) method was performed before method. A comparative study revealed that labellg dex determed sections staed for alone was 1.8 to 6% greater than that determed by sequential double stag method. The difference number per nucleus was not statistically significant, suggestg that double stag method can be used for determg both number labellg dex. This double-stag method was used to study s 12 patients cancer. The mean numbers for positive negative were , respectively. The former number was significantly greater than later (p < ), dicatg that terphase number determed by double-stag method reflects proliferatg activity. Separate enumeration NORs by this method dividg non-dividg cancer is expected to provide important formation for analysis cancer cell ketics. In this study, validity double staig was defed by result that more than 10 counts positive double stag method dicate poor prognosis, on contrary, less than 4 counts negative imply fair prognosis patients cancer. marker proliferatg. In order to demonstrate association between number proliferation cancer carcomas, followg basic study on sequential demonstration s Materials Materials was performed. Methods olorectal carcomas removed by surgery this department were studied. The specimens were fixed 10% formal embedded paraff. The blocks were cut to slices 3 u m thick staed. Immunohistological stag Anti- mouse-monoclonal antibody (P10:DAO) was diluted 100 times used as primary antibody. To sta, DAKO LSAB Kit, Alkale Phosphatase or DAKO LSAB Kit. Peroxidase was used for labelled streptavide-biot (LSAB) method. The LSAB immunoalkale phosphatase method was used New fuchs, Fast red or Fast blue as chromogenic substrate. For LSAB immunoperoxidase method, DAB or AE was used. Introduction The nucleolar organizg regions (NORs) are looped structures rdna occurrg chromosomes Nos. 13, 14, 15, 21, 22. The argyrophilic protes associated NORs are referred to as s. The number, size distribution have been correlated cell's ability to multiply or divide,, as a result, biological malignancy carcomas. The proliferatg cell nuclear antigen (), also called cycl, is a non-histone prote measurg 36kD, which accumulates nuclei durg late G1 to S phase cell cycle. This prote has been known to function as a auxiliary prote DNA polymerase 8, has reby attracted attention as a stag The method Ploton et al." was modified as described below used to sta s before after immunohistochemical stag. Briefly, tissue section was washed distilled water. The reagent prepared dark by combg 1 volume 2% gelat solution 1% formic acid solution 2 volumes 50% silver nitrate solution was spread over section, which was n left at room temperature for varyg times between m to sta s, n washed distilled water.

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5 K YoshidaetaL AStudysequentialdemonstration 18 a positive s red contag numerous black particles b nuclei Fig 1 s negatlve particles staed by double stag method1n s paraff embedded are pale brown nuclei which several seen tissues carcomas x100 の 脚rま5 nqμhεオ8nornμ励εr np瑚ρ 謝yε Table5 Significance results double stag terms survival time 12patients cε1捨 γ髭hご叛π πpハひ4nε8 z距レεcε1捨 n孟hε12pα 8nむ 瞬hD庇5qfcolor8c 01cαncεr n ε q々hε5μ v加1 12patients cluded dead cases 5years after 6 poorprognosissurvivedover5yearsaftersurgely or6 compared lg by dex results ir as not shown l 50f stag number prognoses behavior dex double cancer Over did DNA contrast 1 O analysis negative represented nosis In prognosis biologic parameters comparison fair necessarily reflect positive showed fair labe1 made than5years after surgeσ dead 5 yea岱after 2 poor prog prognosis fair meant ir positive cancel less than40f negative prognosis prognoses NORs number く4 prognosis The patients 10 surge琢 Discussion 50 Survivormore However results double stag coπelated more closely ir prognosis for 一 dex Table5 patients dicated was was lobellg cancer who surgery 枷ε In cancer More than implied lo poor are chromosomes looped structures rdna occurrg Nos The rdna is

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