摘要
目的研究降低DNMT1表达对结直肠癌细胞增殖、迁移及侵袭的影响。方法应用反转录实时荧光定量PCR(qRT-PCR)检测DNMT1在结直肠癌细胞株HCT-8和正常肠上皮细胞株NCM460的表达水平。通过LV-hDNMT1-shRNA慢病毒转染结直肠癌细胞株HCT-8,下调DNMT1在结直肠癌细胞系中表达;细胞功能学实验检测细胞增殖、迁移和侵袭能力;qRT-PCR、Western blot和亚硫酸氢盐甲基化特异性PCR(MSP)方法检测各组细胞中增殖、迁移和侵袭相关指标基因表达水平和甲基化水平。结果结直肠癌细胞HCT-8 DNMT1 mRNA表达水平略高于NCM460。降低HCT-8细胞DNMT1表达,细胞增殖、迁移和侵袭能力增强,Ki-67表达增加,MMP9基因甲基化程度降低其基因表达水平增加、TIMP3基因甲基化程度升高其基因表达降低,差异均具有统计学意义(P<0.05)。结论DNMT1表达降低可促进结直肠癌细胞的HCT-8增殖、迁移和侵袭,其机制可能通过调控MMP9、TIMP3基因甲基化水平影响其蛋白表达,进而对肿瘤细胞侵袭转移产生影响。
Objective To investigate the effect of down-regulating DNMT1 expression on the proliferation,migration and invasion of colorectal cancer cells.Methods The expression of DNMT1 in colorectal cell line HCT-8 and normal intestinal epithelial cell line NCM460 was detected by reverse transcription real-time PCR(qRT-PCR).Colorectal cancer cell line HCT-8 was transfected with LV-hDNMT1-shRNA lentivirus to down-regulate the expression of DNMT1 in colorectal cancer cell line.The cell proliferation,migration and invasion ability were detected by cell function test.qRT-PCR,Western blot and bisulfite methylation-specific PCR(MSP)were used to detect the expression and methylation levels of genes related to proliferation,migration and invasion in each group.Results The expression level of DNMT1 mRNA in HCT-8 cells was slightly higher than that in NCM460 cells.When DNMT1 expression was decreased,the proliferation,migration and invasion ability of HCT-8 cells were enhanced,Ki-67 expression was increased,the methylation degree of MMP9 gene was decreased,the gene expression level was increased,and the methylation degree of TIMP3 gene was increased,and the gene expression was decreased;the differences were statistically significant(P<0.05).Conclusion Decreased expression of DNMT1 can promote the proliferation,migration and invasion of HCT-8 in colorectal cancer cells,and its mechanism may affect the protein expression of MMP9 and TIMP3 by regulating the methylation levels of MMP9 and TIMP3 genes,which may affect the invasion and metastasis of tumor cells.
作者
赵欢
程波
许琳
丁姗姗
吴西钊
刘珊
李志宇
孙锁柱
ZHAO Huan;CHENG Bo;XU Lin;DING Shan-shan;WU Xi-zhao;LIU Shan;LI Zhi-yu;SUN Suo-zhu(Department of Pathology,Rocket Force Special Medical Center,PLA,Beijing100088,China;Department of Basic Medicine Hebei North University,Zhangjiakou 075000,China)
出处
《诊断病理学杂志》
2023年第2期105-109,114,共6页
Chinese Journal of Diagnostic Pathology
基金
国家自然科学基金(31571164)
关键词
DNA甲基转移酶1
结直肠癌
侵袭转移
DNA methyltransferase 1
Colorectal cancer
Invasion and metastasis
