ICG-001对食管鳞癌细胞增殖的抑制作用及其分子机制

Inhibitory effect of ICG-001 on proliferation of esophageal squamous carcinoma cells and its molecular mechanisms

目的:检测小分子抑制剂ICG-001对食管鳞癌细胞增殖的抑制作用并探讨其分子机制。方法:使用50和100μmol/L的小分子抑制剂ICG-001处理食管鳞癌细胞系KYSE410和KYSE450,以溶剂DMSO处理细胞作为对照,进行细胞增殖活力、集落形成能力、细胞周期分布及凋亡检测;分别使用实时荧光定量PCR(qPCR)和Western blot检测ICG-001处理细胞中相关分子(SKP2、Survivin和TCF4)mRNA和蛋白水平的表达变化。结果:与对照组比较,ICG-001处理可显著抑制食管鳞癌细胞系KYSE410和KYSE450的增殖活力和集落形成能力,并可显著诱导细胞发生G0/G1期阻滞(P<0.01);SKP2、Survivin和TCF4的m RNA和蛋白表达水平在ICG-001处理组均显著降低(P<0.01)。结论:ICG-001可显著抑制食管鳞癌细胞的增殖活力和集落形成能力,并可诱导细胞发生G0/G1期阻滞,该作用可能与其抑制β-catenin/TCF4的转录活性及其下游分子Survivin和SKP2的表达有关。

OBJECTIVE:To investigate effects of small molecule inhibitor ICG-001 on malignant phenotypes of esophageal squamous carcinoma(ESCC)cells and to explore its molecular mechanisms.METHODS:The ESCC cell lines KYSE410 and KYSE450 were treated with ICG-001 of 50 and 100μmol/L,or with solvent DMSO as control.Treated cells were evaluated for cell proliferation viability,colony formation ability,cell cycle distribution and apoptosis.mRNA and protein expression of related molecules(SKP2,Survivin,and TCF4)were detected by real-time PCR(qPCR)and Western blot.RESULTS:Compared with the control group,ICG-001 treatment significantly inhibited the proliferation and colony formation ability of KYSE410 and KYSE450,and significantly induced G0/G1 phase arrest(P<0.01).In addition,expression levels of SKP2、Survivin and TCF4 were significantly decreased(P<0.01).CONCLUSION:ICG-001 significantly inhibited the proliferation viability and colony formation ability,and induced G0/G1 phase arrest of the two ESCC cell lines.The mechanisms may be related to the inhibition of b-catenin/TCF4 transcriptional activity and the expression of downstream molecules:Survivin and SKP2.