miR-223通过SEPT6抑制NCK-SOCS7信号通路促进前列腺癌细胞增殖及侵袭

MiR-223 targeting SEPT6 inhibits proliferation and invasion of prostate cancer cells through NCK-SOCS7 signaling pathway

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。

Objective To investigate whether miR-223 targeting SEPT6 regulates NCK- SOCS7 signaling pathway and affects the progression of prostate cancer. Methods MiR 223 mimics （miR MM） and antiscnsc oligonucleotides （miR At）） were used to up regulate and inhibit the ex pression of miR 223 in prostate cancer cell line DU145, respectively, and compared with control vec tot group （miR NC）. Then we used SEPT6 inhibitor sir 1219 and control vector sir NC co trans fcctcd 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 SOCSV, 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 gone, 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.