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. 2003 Aug;9(8):1786-90.
doi: 10.3748/wjg.v9.i8.1786.

Effect of gastrin on differentiation of rat intestinal epithelial cells in vitro

Zhou Wang  1 , Wei-Wen Chen, Ru-Liu Li, Bin Wen, Jing-Bo Sun
Affiliations

Effect of gastrin on differentiation of rat intestinal epithelial cells in vitro

Zhou Wang et al. World J Gastroenterol. 2003 Aug.

Abstract

Aim: To investigate the effect of gastrin on differentiation of IEC-6 cell line in vitro.

Methods: IEC-6 cells were incubated with gastrin. On day 7 after treatment, cell morphology was examined by light microscope, and on day 20, the cellular ultrastructures were examined by electron microscope. After exposure to gastrin for 6 hours, villin mRNA was analyzed by reverse transcription-polymerase chain reaction, and on day 7, the expression of villin was examined by immunocytochemical analysis with laser confocal microscope.

Results: After exposure to gastrin, IEC-6 cells showed differentiated phenotypes as villas enterocytes and contained an abundance of plasma, small nuclei with nucleoli, and were arranged regularly. There were numerous microvilli around edge of the cells, and several cells showed columnar structures. Villin mRNA expression in cytoplasm was increased in comparison with control.

Conclusion: Differentiated characteristics of villus enterocytes and phenotypic changes of rat intestinal epithelial cells (IEC-6) are induced by gastrin, and the effects of gastrin are correlated to increased villin expression.

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Figures

Figure 1
Figure 1
Morphology of IEC-6 cells. a: Gastrin-treated cells(250×) contained an abundance of plasma, small nuclei with nucleoli, and were arranged regularly. b: One of gastrin-treated cells (400×) showed the tendency to form microvilli on the edge(open arrows), and cytoskeleton-like staining in plasma (solid arrows). c: Control cells (250×) contained sparse plasma, large nuclei without nucleoli, and were in irregular arrangement and immature shape. d: One of control cells (400×) showed no tendency to form microvilli on the edge, and nucleus was relatively larger and had no nucleolus.
Figure 2
Figure 2
Ultrastructural changes of IEC-6 cells. a: Gastrin-treated cells (5000×, bar = 1 μm) showed columnar structures(the nuclei were shown by open arrows) with numerous microvilli on the edge (solid arrows). b: Gastrin-treated cells (12000×, bar = 500 nm)developed numerous microvilli (open arrows) and lots of endocytic vesicles appeared under the apical membrane (solid arrows). c: Control cells (5000×, bar = 1 μm) were thin and flat. Relatively large nuclei (open arrows) and scanty plasma were observed. d: Only sparse microvilli (open arrow) and endocytic vesicles (solid arrow) were seen in control cells (12000×, bar = 500 nm).
Figure 3
Figure 3
RT-PCR products from IEC-6 cells on agarose electrophoresis. a: Marker (brighter band: 500 bp), b: Control, c: Gastrin treated cells, d: β-actin(Control), e: β-actin (Gastrin).
Figure 4
Figure 4
Villin expression in IEC-6 cells. a: Gastrin-treated cells (800×), b: Control cells (800×).
See this image and copyright information in PMC

References

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