Protein disulfide isomerase A3: A potential regulatory factor of colon epithelial cells

Yang Li, Z. Huang, Hai-ping Jiang
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Abstract

Aim: This study aimed to investigate the effects of protein disulfide isomerase A3 (PDIA3) on the proliferation and apoptosis of colon epithelial cells, so as to explore its possible role in colon compensation of ultra-short bowel syndrome. Methods: The expression of PDIA3 gene in NCM460 colonic epithelial cells was upregulated and silenced by liposome transient transfection technique. The expression of PDIA3 protein was determined by Western blotting, the proliferation rate of NCM460 cells was detected by CCK8, and the apoptosis rate of NCM460 cells was determined by flow cytometry. Results: DNA sequencing and Western blotting results successfully verified that PDIA3 protein expression in NCM460 cells was upregulated and silenced by liposomal transfection. In the PDIA3 overexpression experiment, the proliferation rate of the experimental group was lower than that of the empty carrier group at 24 h, 48 h, and 72 h, and the apoptosis rate of the experimental group was higher than that of the empty carrier group (P < 0.05, the difference was statistically significant). In the PDIA3-silenced experiment, the proliferation rate of the experimental group was higher than that of the empty carrier group at 24 h, 48 h, and 72 h, and the apoptosis rate of the experimental group was lower than that of the empty carrier group (P < 0.05, the difference was statistically significant). Conclusion: PDIA3 inhibits the proliferation of human colonic epithelial cells (NCM460) and promotes their apoptosis, which may not be a key regulatory protein in colon compensation of ultra-short bowel syndrome.
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蛋白二硫异构酶A3:结肠上皮细胞的潜在调节因子
目的:本研究旨在研究蛋白二硫异构酶A3 (PDIA3)对结肠上皮细胞增殖和凋亡的影响,探讨其在超短肠综合征结肠代偿中的可能作用。方法:采用脂质体瞬时转染技术,上调PDIA3基因在NCM460结肠上皮细胞中的表达并使其沉默。Western blot检测PDIA3蛋白的表达,CCK8检测NCM460细胞的增殖率,流式细胞术检测NCM460细胞的凋亡率。结果:DNA测序和Western blotting结果成功验证了脂质体转染后PDIA3蛋白在NCM460细胞中的表达上调和沉默。在PDIA3过表达实验中,实验组在24 h、48 h、72 h的增殖率低于空载体组,凋亡率高于空载体组(P < 0.05,差异有统计学意义)。pdia3沉默实验中,实验组在24 h、48 h、72 h的增殖率均高于空载体组,凋亡率低于空载体组(P < 0.05,差异有统计学意义)。结论:PDIA3抑制人结肠上皮细胞(NCM460)增殖并促进其凋亡,可能不是超短肠综合征结肠代偿的关键调节蛋白。
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