Evaluation of cell proliferation in rat tissues with BrdU, PCNA, Ki-67(MIB-5) immunohistochemistry and in situ hybridization for histone mRNA
Evaluation of cell proliferation in rat tissues with BrdU, PCNA, Ki-67(MIB-5) immunohistochemistry and in situ hybridization for histone mRNA
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DOI：
10.1177/002215540305101212
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发表时间：
2003-12-01
影响因子：
3.2
通讯作者：
Henderson, EB
中科院分区：
文献类型：
--
作者：
Muskhelishvili, L;Latendresse, JR;Henderson, EB
石蜡包埋组织切片中评估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可以通过IP注射给实验动物，在细胞周期的DNA合成阶段很容易被结合到细胞核中，并可通过抗BrdU抗体进行检测。这种方法有几个缺点，在必要时可以替代BrdU IHC的准确的增殖活性评估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另一种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)组织中抗增殖细胞核抗原、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抗体与组蛋白mRNA Lis、抗BrdU抗体与抗Ki-67Lis抗体的相关性均有统计学意义。抗BrdU抗体与抗PCNALIS抗体之间无明显相关性。这些结果表明，组蛋白mRNA的ISH和MIB-5的IHC是评估大鼠石蜡包埋的更新组织中细胞增殖的较好的技术。他们可以在必要时替代BrdU IHC。
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.