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Published on: July 18, 2019
[Bile salts induce differentiation in cultured human normal esophageal mucosal epithelial cells]
Objective:
To investigate the effects of bile salts on the proliferation and differentiation of human normal esophageal mucosal epithelial cells on cultured.
Methods:
Normal human esophageal mucosa was obtained during operation. The esophageal epithelial cells were isolated, cultured, and treated with 6 different conjugated bile salts [glycocholate (GC), glycochenodeoxycholate (GCDC), glycodeoxycholate (GDC), taurochenodeoxycholate (TCDC), and taurodeoxycholate (TDC), all of the concentration of 50 micromol/L, and taurocholate (TC) of the concentration of 20 micromol/L], and their mixtures the concentration of 50 micromol/L respectively. One, three, and five days later MTT assay was applied to detect the cell proliferation. The cell cycle was assayed by flow cytometry with propidium iodide staining 1 and 3 d after treating with the bile salts. The cytokeratin 13 (CK13) in the differentiated cells and cytokeratin 14 (CK14) in the proliferating cells were detected by immunocytochemical assay. The concentration of intercellular calcium ([Ca(2+)]i) was analyzed by Laser Scanning Confocal Microscope (LSCM) in cells with TC, mixed bile salts and GC.
Results:
The cultured esophageal epithelial cells treated by the bile salts, except those treated by GC for 1 - 3 days, became larger and shuttle-like, with the cell proliferation inhibited, the percentages of cells in G(0)-G(1) phase increased and percentages of cells in S phase decreased. The percentages of CK14 positive cells were increased, but the percentages of CK13 positive cells were decreased time-dependently in cells treated for 1 - 5 days. The most obvious effects were seen in those cells treated with TC. The percentage of CK13 positive cells reached 74% +/- 8% in those cells treated with TC for 5 days, higher significantly than the percentage in the control cell (22% +/- 7%), (P < 0.01). The [Ca(2+)]i increased significantly only several minutes after treatment of TC and mixed bile salts, however, GC failed to cause the increase of [Ca(2+)]i.
Conclusion:
GCDC, GDC, TC, TCDC, TDC and their mixture all induce differentiation of cultured human normal esophageal mucosal epithelial cells, but nor does GC. Increased [Ca(2+)]i is related to the TC-induced differentiation in those esophageal mucosal epithelial cells.
