Original Article
Copyright ©2014 Baishideng Publishing Group Inc.
World J Gastroenterol. Aug 7, 2014; 20(29): 10062-10070
Published online Aug 7, 2014. doi: 10.3748/wjg.v20.i29.10062
Figure 1
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Figure 1 Western blot analysis of calcyclin binding protein/Siah-1 interacting protein expression in gastric cancer cell lines. CacyBP/SIP: Calcyclin binding protein/Siah-1 interacting protein.
Figure 2
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Figure 2 Gastrin can stimulate calcyclin binding protein/Siah-1 interacting protein nuclear translocation. A: Immunofluorescent localization of CacyBP/SIP in cultured gastric cancer cells. a, b and c, unstimulated cells; d, e and f, cells stimulated with gastrin (10-8 mol/L) for 8 h; a and d, immunostained using CacyBP/SIP MAb; b and e, PI stained; c and f, merged images; B: Amounts of CacyBP/SIP in nuclear extracts are shown before and after gastrin (10-8 mol/L) stimulation. C: Cytosolic markers; N: Nuclear markers; PI: Propidium iodide; PARP: Poly ADP-ribose polymerase; CacyBP/SIP: Calcyclin binding protein/Siah-1 interacting protein.
Figure 3
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Figure 3 Calcyclin binding protein/Siah-1 interacting nuclear translocation is able to promote proliferation of gastric cancer cells. A: Effects of calcyclin binding protein/Siah-1 interacting protein (CacyBP/SIP) nuclear translocation on the proliferation of SGC7901 and MKN45 cells. As described in Materials and Methods, the cells were treated with gastrin (10-6-10-10 mol/L) for 24 or 48 h, and their viability was determined by MTT assay. Columns, means; bars, mean ± SD of three separate experiments in which each treatment was done in 5 wells; B: CacyBP/SIP nuclear translocation results in enhanced anchorage-dependent growth. Colony formation in SGC7901 and MKN45 cells with or without 10-8 mol/L gastrin. aP < 0.05 vs control (without gastrin).
Figure 4
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Figure 4 Expression of calcyclin binding protein/Siah-1 interacting protein is evaluated by Western blot in cells stably transfected with calcyclin binding protein/Siah-1 interacting protein siRNAs. β-actin was used as an internal control. CacyBP/SIP: Calcyclin binding protein/Siah-1 interacting protein.
Figure 5
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Figure 5 Localization of calcyclin binding protein/Siah-1 interacting protein nuclear translocation. A: Immunofluorescent localization of CacyBP/SIP in cultured SGC7901-CacyBP/SIPsi1 cells. a, b and c, unstimulated cells; d, e and f, cells 8 h after gastrin (10-8 mol/L) stimulation; a and d, immunostained using anti-CacyBP/SIP MAb; b and e, PI stained; c and f, merged images; B: Amounts of CacyBP/SIP in nuclear extracts are shown before and after gastrin (10-8 mol/L) stimulation. CacyBP/SIP: Calcyclin binding protein/Siah-1 interacting protein; C: Cytosolic markers; N: Nuclear markers.
Figure 6
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Figure 6 Inhibiting calcyclin binding protein/Siah-1 interacting protein nuclear translocation blocks proliferation of gastric cancer cells. A: Effects of gastrin on the viability of SGC7901-CacyBP/SIPsi1 and MKN45-CacyBP/SIPsi1 cells. The cells were treated with gastrin (10-6-10-10 mol/L) for 24 or 48 h, and their viability was determined by MTT assay. Columns, means; bars, mean ± SD of three separate experiments in which each treatment was done in 5 wells; B: Inhibiting CacyBP/SIP expression resulted in the decrease of anchorage-dependent growth and colony formation in SGC7901-CacyBP/SIPsi1 and MKN45-CacyBP/SIPsi1 cells with or without 10-8 mol/L gastrin. CacyBP/SIP: Calcyclin binding protein/Siah-1 interacting protein.