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RNA interference-mediated Gal-3 knockdown inhibits proliferation and promotes apoptosis of pancreatic cancer cells

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Author:
No author available
Journal Title:
World Chinese Journal of Digestology
Issue:
11
DOI:
10.11569/wcjd.v26.i11.648
Key Word:
胰腺癌;半乳糖凝集素-3;增殖;凋亡

Abstract: 目的 探讨RNA干扰半乳糖凝集素-3(galectin-3,Gal-3)表达对胰腺癌细胞增殖和凋亡的影响及其机制.方法 将培养的Panc-1细胞随机分为对照组(未处理)、NC组(转染control-siRNA)和Gal-3干扰组(转染Gal-3-siRNA),以小干扰RNA(Small interfering RNA,siRNA)技术干扰Panc-1细胞中Gal-3表达后,RT-PCR和Western blot检测干扰效果,CCK-8法检测细胞增殖,流式细胞仪检测细胞凋亡,Western blot检测细胞中ki67、细胞周期蛋白Dl(CyclinD1)、活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved caspase-3)抗体和β-连环蛋白(β-catenin)蛋白表达.结果 与对照组相比,NC组中Gal-3 mRNA(0.99±0.08vs 1.01±0.06)和蛋白(0.36±0.03 vs 0.34±0.05)的表达差异不显著(P>0.05),而Gal-3干扰组中Gal-3mRNA(0.38±0.02 vs 1.01±0.06)和蛋白(0.10±0.01vs 0.34±0.05)的表达均显著降低(P<0.05);Gal-3干扰组细胞增殖能力减弱(24 h:0.55±0.03 vs 0.71±0.05;48 h:0.76±0.05 vs 0.97±0.06;72 h:1.08±0.06 vs1.32±0.09),G0/G1期细胞百分比升高(79.48±1.32 vs71.52±1.15),S期(14.26±1.08 vs 18.24±1.06)和G2/M期(6.21±0.78 vs 10.19±1.52)降低,凋亡能力增强(13.26±2.28 vs 5.82±0.35),与对照组相比,差异均有统计学意义(P<0.05);与对照组相比,Gal-3干扰组中ki67(0.24±0.02 vs 0.96±0.07)、CyclinD1(0.26±0.03vs 0.88±0.09)和β-catenin(0.42±0.05 vs 0.87±0.05)蛋白的表达水平均明显降低,而Cleaved Caspase-3(0.70±0.06 vs 0.32±0.03)蛋白表达水平明显升高,差异有统计学意义(P<0.05);NC组与对照组间各指标差异均无统计学意义(P>0.05).结论 RNA干扰Panc-1细胞中Gal-3表达,可抑制细胞增殖并促进细胞凋亡,其机制可能与抑制Wnt/β-catenin信号通路有关.

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