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[Effect of hepatocyte growth factor on proliferation of cultured human eccrine sweat gland epithelial cells]
Xia Lei1, Jinjin Wu, Yuangang Lu
1Department of Dermatology, Daping Hospital, Third Military Medical University, Chongqing, 400042, P.R. China.
Objective:
To investigate the effect of hepatocyte growth factor (HGF) on proliferation of cultured human eccrine sweat gland epithelial cells (hESGc) and the involvement of phosphorylation of ERK1/2.
Methods:
hESGc were cultured in keratinocyte serum free medium (KSFM) and the first generation of hESGc was harvested. The expression of C-met was detected by immunocytochemistry. MTT assay was used to detect the effect of HGF on the proliferation of hESGc. The cells were divided into blank group, control group and experimental group. The culture density was 2 x 10(3) cells/hole in control group and experimental group. Two hundred microL KSFM with HGF in different levels was added to every hole. hESGc were cultured in KSFM with HGF at different levels (2, 20, 40 and 80 ng/mL) in experimental group, in KSFM without HGF in control group, and in KSFM without HGF and no hESGc in blank group. The cell proliferation was observed in experimental group 2 and 4 days later. Western blot was used to detect the expression of phosphorylated ERK1/2 at 40 ng/mL HGF after 0, 5, 30, 90 and 120 minutes.
Results:
The results were positive for anti-C-met staining in the cytoplasm. HGF (40 ng/mL and 80 ng/mL) significantly improved the proliferation of hESGc (P < 0.05). When cultured in the KSFM with 40 ng/mL HGF, the cell proliferation rate and the absorbance were 74.2%, 0.2393 +/- 0.0709 at 2 days and 74.8%, 0.2878 +/- 0.0743 at 4 days; showing significant differences when compared with control group (P < 0.05). When cultured in KSFM with 80 ng/mL HGF, the cell proliferation rate and the absorbance were 54.5%, 0.2123 +/- 0.0592 at 2 days and 40.3%, 0.2310 +/- 0.0567 at 4 days; showing significant differences when compared with control group (P < 0.05). The expression of p-ERK1/2 reached to the maximum after stimulation of 40 ng/mL HGF for 5 minutes, and relative integral absorbance (RIA) was 0.5932 +/- 0.1922, increased 8.1 times compared with instant stimulation (P < 0.01).
Conclusion:
HGF could induce the proliferation of hESGc and activate the phosphorylation of ERK 1/2 protein.
