【摘要】 目的探讨miR-223是否通过GTP结合蛋白基因SEPT6调控NCK-SOCS7信号通路并影响前列腺癌进展。方法通过miR-223模拟物(miR-MM)及其反义寡核苷酸(miR-AO)分别上调及抑制前列腺癌细胞系DU145中miR-223的表达,并与miR-223空载体组(miR-NC)进行比较。通过SEPT6抑制剂siR-1219及空白载体siR-NC分别与miR-NC共转染DU145细胞,同样再分别与miR-AO共转染DU145细胞。利用QPCR检测miR-223、SEPT6、NCK和SOCS7基因的表达,MTT法及Transwell小室法分别进行细胞增殖和侵袭实验。结果在miR-MM组,提高miR-223表达,能够显著抑制SEPT6、NCK及SOCS7表达水平,并且促进细胞增殖及侵袭力[细胞数为(103±37)个];反之在miR-AO组,则SEPT6、NCK及SOCS7基因表达显著增高,细胞增殖及侵袭力明显减弱[细胞数为(29±11)个]。进一步研究发现,在miR-NC+siR-1219组,miR-223及SEPT6表达分别较高和较低,NCK及SOCS7与SEPT6表达趋势一致,也较低,此时细胞增殖及侵袭能力较强[细胞数为(63±25)个];而在miR-AO+siR-NC组,miR-223及SEPT6表达分别较低和较高,NCK及SOCS7与SEPT6表达趋势也一致,也较高,而细胞增殖及侵袭能力较弱[细胞数为(21±6)个](P<0.05)。结论 miR-223能够抑制SEPT6基因表达,导致下游DNA损伤修复信号通路NCK-SOCS7被抑制,从而增加前列腺癌的恶性程度。本研究可能为前列腺癌的进展及潜在的治疗靶点提供基础研究依据。更多还原

【Abstract】 Objective To investigate whether miR-223 targeting SEPT6 regulates NCKSOCS7 signaling pathway and affects the progression of prostate cancer. Methods MiR-223 mimics(miR-MM)and antisense oligonucleotides(miR-AO)were used to up-regulate and inhibit the expression of miR-223 in prostate cancer cell line DU145,respectively,and compared with control vector group(miR-NC).Then we used SEPT6 inhibitor siR-1219 and control vector siR-NC co-transfected DU145 cells with miR-NC and miR-AO.The expression of miR-223,SEPT6,NCK and SOCS7 genes were detected by QPCR,and cell proliferation assay and invasion assay were performed by MTT assay and Transwell chamber,respectively. Results In the miR-MM group,increased expression of miR-223 significantly inhibited the expression levels of SEPT6,NCK and SOCS7,and promoted cell proliferation rate and invasiveness(103±37).Conversely,in group of miR-AO,SEPT6,NCK and SOCS7 gene expression increased significantly,cell proliferation rate and invasiveness were significantly reduced(29±11).Further studies showed that the expressions of miR-223 and SEPT6 were higher and lower in miR-NC+siR-1219 group,respectively,and the expression trends of NCK and SOCS7 were the same as those of SEPT6.The cell proliferation rate and invasion ability in miR-NC+siR-1219 group were higher(63±25).In the miR-AO+siR-NC group,the expressions of miR-223 and SEPT6 were lower and higher,respectively,and the expression trends of NCK,SOCS7 and SEPT6 were also consistent,while the cell proliferation rate and invasion ability were lower(all P<0.05). Conclusions MiR-223 can inhibit the expression of SEPT6 gene,leading to the inhibition of downstream DNA damage repair signaling pathway NCK-SOCS7,thereby increasing the progression of prostate cancer.This study may provide a basic research for new information of the progression prostate cancer and its potential therapeutic targets.更多还原