目的探讨超声微泡介导miR-545-3p调节HOXA3对结直肠癌(colorectal cancer,CRC)细胞增殖、凋亡、侵袭的影响。方法采用实时荧光定量聚合酶链式反应(quantitative real time polymerase chain reaction,qRT-PCR)检测SW620、SW480、LoVo、C...目的探讨超声微泡介导miR-545-3p调节HOXA3对结直肠癌(colorectal cancer,CRC)细胞增殖、凋亡、侵袭的影响。方法采用实时荧光定量聚合酶链式反应(quantitative real time polymerase chain reaction,qRT-PCR)检测SW620、SW480、LoVo、CACO-2及正常结肠上皮细胞系NCM460中miR-545-3p、同源盒基因3(homeobox geneA3,HOXA3)mRNA表达水平;将生长较好的SW480细胞分为空白组、微泡组、miR组、过表达miR-545-3p组、微泡+miR组、微泡+过表达miR-545-3p组、微泡+过表达miR-545-3p+空载体(pcDNA6.2/nLumio-DEST,pcDNA)组、微泡+过表达miR-545-3p+HOXA3组,采用qRT-PCR法检测各组miR-545-3p和HOXA3表达,采用MTT检测细胞增殖,采用流式细胞术检测细胞凋亡,采用Transwell实验检测细胞侵袭,采用Western blotting检测HOXA3、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)、金属基质蛋白酶9(matrix metalloproteinase,MMP-9)、(Bcl-2-associated X protein,Bax)蛋白表达水平。通过皮下注射SW480细胞悬液,建立裸鼠移植瘤模型,并分为对照组、微泡组、miR微泡组、miR-545-3p微泡组,每组10只,检测裸鼠移植瘤瘤重。结果CRC细胞中miR-545-3p表达下调,HOXA3 mRNA表达上调(P<0.05),以SW480细胞变化最为显著,故作为后续研究对象。过表达miR-545-3p与HOXA3-WT共转染的荧光素酶活性低于miR NC与HOXA3-WT共转染(P<0.05)。过表达miR-545-3p组可上调miR-545-3p表达,下调HOXA3表达,抑制细胞增殖、侵袭数及PCNA、MMP-9蛋白表达,促进细胞凋亡及Bax蛋白表达(P<0.05),微泡+过表达miR-545-3p组上调miR-545-3p表达,下调HOXA3表达,抑制细胞增殖、侵袭数及PCNA、MMP-9蛋白表达,促进细胞凋亡及Bax蛋白表达效果优于过表达miR-545-3p组(P<0.05)。miR-545-3p微泡组裸鼠移植瘤瘤重显著低于对照组、微泡组、miR微泡组(P<0.05)。结论超声微泡介导miR-545-3p可通过下调HOXA3抑制CRC细胞增殖、侵袭,诱导其凋亡,为临床治疗CRC提供新的给药方式及潜在靶点,为CRC分子靶向治疗技术提供依据。展开更多
The intricacies of Alzheimer’s disease pathogenesis are being increasingly illuminated by the exploration of epigenetic mechanisms,particularly DNA methylation.This review comprehensively surveys recent human-centere...The intricacies of Alzheimer’s disease pathogenesis are being increasingly illuminated by the exploration of epigenetic mechanisms,particularly DNA methylation.This review comprehensively surveys recent human-centered studies that investigate whole genome DNA methylation in Alzheimer’s disease neuropathology.The examination of various brain regions reveals distinctive DNA methylation patterns that associate with the Braak stage and Alzheimer’s disease progression.The entorhinal cortex emerges as a focal point due to its early histological alterations and subsequent impact on downstream regions like the hippocampus.Notably,ANK1 hypermethylation,a protein implicated in neurofibrillary tangle formation,was recurrently identified in the entorhinal cortex.Further,the middle temporal gyrus and prefrontal cortex were shown to exhibit significant hypermethylation of genes like HOXA3,RHBDF2,and MCF2L,potentially influencing neuroinflammatory processes.The complex role of BIN1 in late-onset Alzheimer’s disease is underscored by its association with altered methylation patterns.Despite the disparities across studies,these findings highlight the intricate interplay between epigenetic modifications and Alzheimer’s disease pathology.Future research efforts should address methodological variations,incorporate diverse cohorts,and consider environmental factors to unravel the nuanced epigenetic landscape underlying Alzheimer’s disease progression.展开更多
文摘The intricacies of Alzheimer’s disease pathogenesis are being increasingly illuminated by the exploration of epigenetic mechanisms,particularly DNA methylation.This review comprehensively surveys recent human-centered studies that investigate whole genome DNA methylation in Alzheimer’s disease neuropathology.The examination of various brain regions reveals distinctive DNA methylation patterns that associate with the Braak stage and Alzheimer’s disease progression.The entorhinal cortex emerges as a focal point due to its early histological alterations and subsequent impact on downstream regions like the hippocampus.Notably,ANK1 hypermethylation,a protein implicated in neurofibrillary tangle formation,was recurrently identified in the entorhinal cortex.Further,the middle temporal gyrus and prefrontal cortex were shown to exhibit significant hypermethylation of genes like HOXA3,RHBDF2,and MCF2L,potentially influencing neuroinflammatory processes.The complex role of BIN1 in late-onset Alzheimer’s disease is underscored by its association with altered methylation patterns.Despite the disparities across studies,these findings highlight the intricate interplay between epigenetic modifications and Alzheimer’s disease pathology.Future research efforts should address methodological variations,incorporate diverse cohorts,and consider environmental factors to unravel the nuanced epigenetic landscape underlying Alzheimer’s disease progression.
