Use of in situ detection of histone mRNA in the assessment of epidermal proliferation: comparison with the Ki67 antigen and BrdU incorporation

Abstract

The labelling index is commonly used as a measure of proliferation. However, the use of tritiated thymidine or BrdU labelling of S-phase cells is limited to prospective samples. We have employed an oligonucleotide cocktail complementary to the mRNA species encoding the replication-dependent histones H2B, H3 and H4 for non-isotopic in situ hybridization (NISH), and have compared the resultant proliferation indices in normal skin with those obtained by bromodeoxyuridine (BrdU) incorporation and by Ki67 immunohistochemistry (IHC) using the monoclonal antibody MIB1. In addition, we compared the staining characteristics of histone NISH and Ki67 IHC in a further 25 samples from a variety of inflammatory dermatoses and neoplastic conditions, as well as from normal skin. In normal skin, S-phase (histone NISH and BrdU) and cycling (Ki67) cells were confined to the basal and low suprabasal layers. The labelling indices determined by histone NISH and BrdU incorporation were similar, whereas that of Ki67 IHC was four times greater. In biopsies from hyperproliferative dermatoses and dysplastic or malignant lesions, the number of histone NISH- and Ki67 IHC-positive cells was generally elevated; in accordance with the differential expression of these two markers during the cell cycle, MIB1 consistently gave higher results. The advantage of histone NISH over Ki67 IHC is that it is a marker of the same part of the cell cycle as BrdU incorporation. However, the combined use of both histone NISH and Ki67 IHC to measure two cell cycle parameters, namely S-phase and the number of cycling cells, allows more detailed retrospective study of epidermal proliferation than has been possible previously.

Proliferation and phenotype regulation in the subventricular zone during experimental allergic encephalomyelitis: In vivo evidence of a role for nerve growth factor

“…2). In other proliferating cell populations, like epithelial cells or different tumor cells, a similar ratio between markers for the S phase alone (i.e., [ 3 H]thymidine incorporation), and proliferation-associated markers, including Ki67, has been described (17). Evaluation of Ki67 immunoreactivity revealed a doubling of the mitotic index during EAE, compared with both control and adjuvant-injected rats (Fig.…”

Predicting tumor metastasis in patients with oral cancer by means of the proliferation marker Ki67.

“…Recent immunohistochemical studies have been useful for the detection of proliferating cells in tissue samples using polyclonal or monoclonal antibodies against the proliferating cell nuclear antigens (1-4), P53 (5,6) and Ki67 (4)(5)(6)(7)(8). The immunohistochemistry of Ki67 was originally limited to frozen sections, however, as a result of recent developments of the MIB-1 and MIB-3 antibodies, it is now available in paraffin-embedded specimens (6,9,10). The antigen retrieval technique using autoclaving or microwave treatment prior to immunostaining also proves beneficial to retrospective studies.…”

Contrasting Expression Patterns of Histone mRNA and microRNA 760 in Patients with Gastric Cancer

“…Histone mRNAs are tightly regulated during the cell cycle, allowing the synthesis of histone proteins to occur coordinately with the replication of DNA. In malignant tumors, upregulation of histone mRNA indicates proliferative activity of tumor cells, and many studies have demonstrated that histone mRNA accumulates in tumors (14)(15)(16). In most previous studies, histone levels have been measured by only considering H3 core histone levels.…”