摘要
目的探讨赖氨酸特异性组蛋白去甲基化酶1（lysine-specific demethylase 1, LSD1）对结肠癌细胞侵袭转移能力的影响及其机制。方法利用RNAi技术和单胺氧化酶抑制剂（Pargyline）在体外抑制结肠癌细胞（SW620）中LSD1的表达，采用MTT细胞增殖实验、Transwell细胞侵袭实验、FCM细胞凋亡实验检测抑制LSD1的表达对结肠癌细胞SW620增殖、侵袭、凋亡的影响。结果体外合成了三段siRNA序列（siRNA-1554、siRNA-705、siRNA-1973），经检测siRNA-705的沉默效率最高。利用siRNA-705和单胺氧化酶抑制剂体外干预SW620细胞，能明显抑制SW620细胞的增殖、侵袭和转移能力，并能诱导细胞凋亡。同时我们发现，使用siRNA-705和单胺氧化酶抑制剂处理72 h后，结肠癌细胞的E钙黏蛋白的表达水平明显上调，N钙黏蛋白的表达水平明显下调（P〈0.05）。结论体外抑制LSD1的表达能抑制结肠癌细胞的增殖、侵袭转移能力，诱导细胞凋亡，并能上调E钙黏蛋白的表达，下调N钙黏蛋白的表达，三者可能在结肠癌的侵袭转移过程中发挥了重要作用。
ObjectiveTo investigate the effect and molecular mechanisms of LSD1 on proliferation and metastasis of colon cancer.
MethodsThe influence of down-regulated LSD1 expression on proliferation, invasion and apoptosis of colon cancer cells were detected by transwell invasion assay, cell proliferation assay and apoptosis assay, respectively.
ResultsThree independent siRNAs targeting LSD1 （siRNA-1554, siRNA-705, and siRNA-1973） were transfected to SW620 cells to detect gene-silencing efficiency, and the result showed that the knockdown effect of siRNA-705 were better than the other two siRNAs at both mRNA and protein levels. Using siRNA-705 and pargyline （2.5 mmol/L）, we performed transwell invasion assay, cell proliferation assay and apoptosis assay in SW620 cell lines, and found the significant suppression of invasion and growth. Cell apoptosis were induced by siRNA and pargyline （P〈0.05）. Interestingly, up-regulation of E-cadherin and down-regulation of N-cadherin were observed after treated with siRNA-705 and pargyline for 72 hours.
ConclusionInhibition of LSD1 could impair proliferation and invasiveness, and induce apoptosis of colon cancer cells in vitro. It leads to up-regulation of E-cadherin and down-regulation of N-cadherin. All of above may play important roles in invasion and metastasis of colon cancer.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2017年第10期743-748,共6页
National Medical Journal of China
基金
国家自然科学基金（81360366,81302169）
贵州省科学技术基金（黔科合J字[2013]2178号）
贵州省社会发展攻关项目（黔科合SY字[2014]3023号）
