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. 2003 Sep;163(3):969-78.
doi: 10.1016/S0002-9440(10)63456-6.

"VSports手机版" Differential expression of cyclin D1 in the human hair follicle

Xiaowei Xu  1 , Stephen Lyle, Yaping Liu, Benjamin Solky, George Cotsarelis
Affiliations

Differential expression of cyclin D1 in the human hair follicle

Xiaowei Xu et al. Am J Pathol. 2003 Sep.

Abstract

The proliferation of keratinocytes in the hair follicle varies from slowly cycling, intermittently proliferating stem cells in the bulge to rapidly proliferating, transient cells in the bulb. To better understand the biological differences between these two compartments, we sought to identify differentially expressed genes using cDNA macroarray analysis. Cyclin D1 was one of 13 genes increased in the bulge compared to the bulb, and its differential expression was corroborated by quantitative real-time polymerase chain reaction (PCR) on the original samples. Using immunohistochemical staining, laser-capture microdissection (LCM) and quantitative real-time PCR, we localized cyclin D1 to the suprabasal cells of the telogen bulge and anagen outer root sheath (ORS). Surprisingly, cyclin D1, D2, and D3 were not detectable by immunohistochemistry in the rapidly proliferating hair-producing cells of the anagen bulb (matrix cells), while these cells were strongly positive for Ki-67 and retinoblastoma protein. In contrast, pilomatricoma, a tumor thought to be derived from matrix cells, was positive for cyclin D1, D2, and D3. Our results suggest that cyclin D1 may mediate the proliferation of stem cells in the bulge to more differentiated transient amplifying cells in the suprabasal ORS. In contrast, non-cyclin D1-proteins appear to control cell division of the highly proliferative bulb matrix cells VSports手机版. This non-cyclin D1-mediated proliferation may provide a protective mechanism against tumorigenesis, which is overridden in pilomatricomas. Our data also demonstrate that the combination of DNA macroarray, LCM and quantitative real-time PCR is a powerful approach for the study of gene expression in defined cell populations with limited starting material. .

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Figures

Figure 1.
Figure 1.
Images of entire epithelial component of telogen (A) and anagen (B) hair follicles released from human scalp tissue at the basement membrane by incubation with the enzyme dispase (×20).
Figure 2.
Figure 2.
Quantitative real-time PCR demonstrates increased expression of cyclin D1 in telogen follicle. A: Concentration curve of GAPDH. B: Concentration curve of cyclin D1. C: Comparison of cyclin D1 gene expression in telogen bulge and anagen bulb.
Figure 3.
Figure 3.
Immunohistochemical detection of cyclin D1 and Ki67 in telogen bulge (A and B) and anagen ORS (C and D). A: Cyclin D1-positive cells are located in the suprabasal layer of the telogen bulge (arrow, ×100). B: Majority of Ki67-positive cells are located in the suprabasal layer of the telogen bulge (arrowhead, ×100). Cyclin D1-positive (arrow, C) and Ki67-positive cells (arrowheads, D) are located in the suprabasal layer of the anagen ORS (×630).
Figure 4.
Figure 4.
Immunohistochemical detection of cyclin D1, Ki67, and RB in the anagen hair bulb. A: H&E-stained tissue section of anagen hair bulb (DP, dermal papilla; M, matrix; ×200). Higher magnification view of boxed area is shown in D. B: Immunohistochemical staining for cyclin D1 in anagen bulb (×400). The anagen bulb is rich in pigment-laden melanocytes. No true nuclear staining is present. The boxed area is shown in E. C: Negative control of consecutive section of anagen bulb shown in B. The pigment-laden melanocytes are easily identified (×400). The boxed area is shown in F. Arrows in D, E, and F point to pigment-laden cells. G: At anagen onset, the suprabasal cells of the telogen bulge are positive for cyclin D1 (arrowheads). However, the newly formed anagen bulb is negative for cyclin D1 (arrow, ×200). H: Ki67 staining shows that cells around the dermal papilla are strongly positive and proliferating (×200). E: Immunostaining for Rb shows that the matrix cells retain expression of Rb (×200). Immunohistochemical sections are counterstained with hematoxylin.
Figure 5.
Figure 5.
Laser-capture microdissection for isolation of hair follicle basal cells (A to C) and suprabasal cells (D to F). A and D: Overview of the tissue section before the dissection (×40 and ×200). B and E: Overview of the tissue section after the microdissection (×200). C and F: Microdissected tissue on the transfer film carrier (×200). The sections were stained with H&E.
Figure 6.
Figure 6.
Differential expression of cyclin D1 mRNA in ORS. A: Concentration curve of ribosomal 18s. B: Concentration curve of cyclin D1. C: Concentration of cyclin D1 in basal and suprabasal cells relative to concentration in basal layer cells.
Figure 7.
Figure 7.
Pilomatricomas express cyclins D1, D2, and D3. A: H&E stain of pilomatricoma. Arrow indicates shadow cells; arrowhead indicates basoloid cells (×200), Tumor cells are focally positive for cyclin D1 (B), cyclin D2 (C), and cyclin D3 (D) (arrows, ×200).
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References

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