miR―1203和HOTAIRrs7958904位点对乳腺癌细胞生长影响.docVIP

miR―1203和HOTAIRrs7958904位点对乳腺癌细胞生长影响 　　[摘要] 目的 探?长链非编码RNA HOTAIR多态位点rs7958904的遗传变异与miR-1203的关系及两者在乳腺癌细胞生长中的作用。 方法 分别将HOTAIR rs7958904野生型和突变型克隆进双荧光素酶报告载体和过表达载体，双荧光素酶报告载体分别和miR-1203模拟物共转染细胞，检测荧光素酶的表达。过表达载体、miR-1203模拟物或抑制物单独或共转染乳腺癌细胞，荧光定量PCR检测HOTAIR表达，BrdU检测细胞增殖，Annexin V-FITC/PI检测细胞凋亡，Transwell法检测细胞侵袭。 结果 HOTAIR突变型增强了miR-1203对其的靶向结合；miR-1203可以下调突变型HOTAIR的表达，差异有高度统计学意义（P 0.05）。野生型HOTAIR明显促进乳腺癌细胞的增殖侵袭和抑制其凋亡，突变型HOTAIR的上述作用明显减弱且可以被miR-1203阻断，差异均有高度统计学意义（P 0.01）；miR-1203具有与HOTAIR相反的作用，抑制miR-1203表达后，突变型HOTAIR具有与野生型HOTAIR类似的效果。 结论 HOTAIR rs7958904的突变型能下调HOTAIR的功能，此作用可能是增加了miR-1203的结合位点导致的。此发现将为lncRNA SNP功能的研究探讨提供新的研究方向。 　　[关键词] 长链非编码RNA；HOTAIR；乳腺癌；miR-1203；单核苷酸多态性 　　[中图分类号] R737.9 [文献标识码] A [文章编号] 1673-7210（2018）01（c）-0008-05 　　[Abstract] Objective To investigate the association between genetic variation of lncRNA HOTAIR SNP rs7958904 and miR-1203， as well as to explore their roles in the growth of breast cancer （BC） cells. Methods The wild type and mutant HOTAIR SNP rs-7958904 were both cloned into the Dual-Luciferase Reporter Vector and Over-expression vector， respectively. The Dual-Luciferase Reporter Vectors and miR-1203 mimics were co-transfected into the cells， and then the luciferase expression was detected. Furthermore， BC cells were transfected with the overexpression vector and/or miR-1203 mimics/inhibitors. And then， HOTAIR expression was detected by fluorescent quantitative PCR， cell proliferation was assessed by BrdU， cell apoptosis was tested by Annexin V-FITC/PI， and cell invasion was examined by Transwell Assay. Results The binding of miR-1203 and HOTAIR was enhanced by mutant HOTAIR； miR-1203 could down-regulate the expression of mutant HOTAIR （P 0.05）. The wild-type HOTAIR promoted the proliferation and invasion of BC cells significantly but inhibit their apoptosis （P 0.01）； whereas such effect of the mutant HOTAIR declined obviously and could be blocked by miR-1203. miR-1203 had the opposite effect of HOTAIR； when the expression of miR-1203 was in