摘要
目的探讨lnc RNA-ANCR与组蛋白甲基转移酶EZH2的表达及其与结直肠癌细胞侵袭与转移的关系。方法培养结肠癌SW620细胞系,转染ANCR-siRNA至细胞,采用实时荧光定量PCR(RT-PCR)检测ANCR和EZH2表达,Western blot检测EZH2蛋白表达水平,采用RNA pull-down和免疫共沉淀的方法检测ANCR与EZH2的相互作用关系,Transwell实验和划痕实验检测细胞侵袭和转移能力。结果结肠癌SW620细胞中ANCR和EZH2的表达均显著高于正常结肠上皮细胞(P<0.05)。转染ANCR-siRNA的SW620细胞中ANCR的表达较转染阴性对照质粒的SW620细胞显著降低,且EZH2的mRNA和蛋白表达水平均显著降低(P<0.05)。RNA pull-down与免疫共沉淀的结果显示ANCR可与EZH2特异性结合。而Transwell实验和划痕实验显示转染ANCR-siRNA的SW620细胞的侵袭和迁移能力均显著低于转染阴性对照质粒的SW620细胞(P<0.05)。结论结直肠癌细胞中ANCR与EZH2的表达均上调,ANCR可能通过特异性结合EZH2调节结直肠癌细胞的侵袭和转移。
Objective To investigate the expression of anti-differentiation noncoding RNA （lncRNA-ANCR） and histone methyhrans- ferase enhancer of zeste homolog 2 （EZH2） and their relationship with the invasion and metastasis of coloreetal cancer cells. Methods Colorectal cancer cell line SW620 was cultured and transfected with ANCR-siRNA. The ANCR and EZH2 expression were detected by RT-PCR （real-time PCR） and the expression level of EZH2 protein was detected by Western blot. RNA pull-down and co-immunopre- cipitation were used to detect the interaction between ANCR and EZH2. The Transwell assay and scratch test were applied to detect the cell ability of invasion and metastasis. Results The expression of ANCR and EZH2 were higher in SW620 cell than in normal colonic epithelial cell （P〈 0.05）. Compared with SW620 cells transfeeted with negative control plasmid, the expression level of ANCR decreased in SW620 cells transfected with ANCR-siRNA. Moreover, the mRNA and protein expression of EZH2 were decreased （all P〈 0.05）. RNA pull-down and eo-immunoprecipitation results indicated that ANCR could specifically bind to EZH2. Transwell assay and scratch test showed that the invasion and migration ability of SW620 ceils transfected with ANCR-siRNA were significantly attenuated （P 〈 0.05）. Conclusion The ex- pression levels of ANCR and EZH2 in colorectal cancer were significantly upregulated, and ANCR can specifically bind to EZH2 to regulate the invasion and metastasis process of colorectal cancer cells.
出处
《肿瘤药学》
CAS
2017年第2期169-174,共6页
Anti-Tumor Pharmacy
