Muskhelishvili Levan, Latendresse John R, Kodell Ralph L, Henderson Eric B
Charles River Laboratories, Jefferson, Arkansas 72079, USA.
J Histochem Cytochem. 2003 Dec;51(12):1681-8. doi: 10.1177/002215540305101212.
The standard method for assessment of cell proliferation in paraffin-embedded tissue sections is 5-bromodeoxyuridine (BrdU) immunohistochemistry (IHC). BrdU can be administered to laboratory animals via IP injections, is readily incorporated into nuclei during the DNA synthetic phase of the cell cycle, and is detected with an anti-BrdU antibody. This method has several disadvantages, and an accurate method for evaluation of proliferative activity that can substitute for BrdU IHC, when necessary, is of great interest to investigators. Alternative methods for detection of proliferating cells in tissue sections are proliferating cell nuclear antigen (PCNA) IHC, Ki-67 IHC, and in situ hybridization (ISH) for histone mRNA. To determine the optimal choice, we analyzed the correlation of anti-PCNA, anti-Ki-67(MIB-5), and histone mRNA labeling indices (LIs) with anti-BrdU LI in rat highly replicative (renewing) tissues. The correlation between anti-BrdU and histone mRNA LIs, as well as the correlation between anti-BrdU and anti-Ki-67 LIs, was statistically significant. There was no significant correlation between anti-BrdU and anti-PCNA LIs. These results suggest that both ISH for histone mRNA and IHC with MIB-5 are preferable techniques for assessment of cell proliferation in rat paraffin-embedded renewing tissues compared to PCNA IHC. They can substitute for BrdU IHC when necessary.
评估石蜡包埋组织切片中细胞增殖的标准方法是5-溴脱氧尿苷（BrdU）免疫组织化学（IHC）。BrdU可通过腹腔注射给予实验动物，在细胞周期的DNA合成期很容易掺入细胞核，并使用抗BrdU抗体进行检测。该方法有几个缺点，一种在必要时可替代BrdU IHC的准确评估增殖活性的方法引起了研究人员的极大兴趣。检测组织切片中增殖细胞的替代方法是增殖细胞核抗原（PCNA）IHC、Ki-67 IHC和组蛋白mRNA的原位杂交（ISH）。为了确定最佳选择，我们分析了大鼠高复制（更新）组织中抗PCNA、抗Ki-67（MIB-5）和组蛋白mRNA标记指数（LIs）与抗BrdU LI的相关性。抗BrdU与组蛋白mRNA LIs之间的相关性以及抗BrdU与抗Ki-67 LIs之间的相关性具有统计学意义。抗BrdU与抗PCNA LIs之间无显著相关性。这些结果表明，与PCNA IHC相比，组蛋白mRNA的ISH和MIB-5的IHC都是评估大鼠石蜡包埋更新组织中细胞增殖的更优技术。必要时，它们可以替代BrdU IHC。
