miR-572通过靶向调控WWOX影响肺癌细胞恶性生物学行为

Influence of miR-572 on the malignant biological behavior of lung cancer cells via targeting WWOX regulation

目的探讨miR-572靶向氧化还原酶的WW结构域(WW domain containing oxidoreductase,WWOX)调控肺癌细胞增殖、凋亡的分子机制。方法选取50例2016年3月—2018年5月十堰市太和医院手术切除肺癌组织及其相应的癌旁组织。利用脂质体转染技术将miR-572 inhibitor和miR-572 mimics转染至肺癌A549和L9981细胞;采用qRT-PCR检测miR-572和WWOX的mRNA表达水平,MTT法检测细胞增殖能力,流式细胞仪检测细胞周期和细胞凋亡情况。通过生物信息学软件Targetscan分析miR-572的靶基因,荧光素酶报告基因实验检验miR-572和WWOX的靶向关系。结果miR-572在肺癌组织和细胞中高表达(均P<0.05)。下调miR-572后,肺癌细胞A549、L9981中miR-572表达水平均降低(均P<0.05),细胞增殖能力也降低(均P<0.05),细胞G0/G1期比例和细胞凋亡率均升高(均P<0.05);而上调miR-572后,肺癌A549、L9981细胞增殖能力升高(均P<0.05),细胞G0/G1期比例和细胞凋亡率则降低(均P<0.05)。WWOX在肺癌组织和细胞中低表达(均P<0.05),且在肺癌组织中WWOX与miR-572表达呈负相关(r=-0.669,P<0.001)。下调WWOX可逆转下调miR-572对肺癌细胞的增殖抑制、周期阻滞和凋亡促进作用。结论下调miR-572可抑制肺癌细胞增殖并诱导细胞凋亡,其作用机制可能与靶向负调控WWOX有关。

Objective To investigate the molecular mechanism of miR-572 targeting WW domain containing oxidoreductase(WWOX)in regulating the proliferation and apoptosis of lung cancer cells.Methods A total of 50 cases of lung cancer tissues and their corre-sponding adjacent tissues of lung cancer patients who underwent surgical resection in Taihe Hospital of Shiyan Gity from March 2016 to May 2018 were collected.miR-572 inhibitor and miR-572 mimics were transfected into lung cancer A549 and L9981 cells by using liposome transfection technology.The expressions of miR-572 and WWOX mRNA were determined by qRT-PCR,the cell proliferation was measured by MTT assay,and the cell cycle and apoptosis were measured by flow cytometry.Targetscan was used to analyze the target genes of miR-572,the luciferase reporter gene experiment confirmed the targeting relationship between miR-572 and WWOX.Results miR-572 was highly expressed in lung cancer tissues and cells(all P<0.05).After down-regulating miR-572,the proliferation ability of lung cancer A549 and L9981 cells decreased(all P<0.05),but the cell G0/G1 phase ratio and apoptosis rate increased(all P<0.05).However,after up-regulating miR-572,the proliferation ability of miR-572 in lung cancer A549 and L9981 cells increased(all P<0.05),while the cell G0/G1 phase ratio and apoptosis rate of cells decreased(all P<0.05).WWOX was low expressed in lung cancer tissues and cells(all P<0.05),and it was negatively correlated with the expression of miR-572 in lung cancer tissues(r=-0.669,P<0.001).Down-regulating the expression of WWOX could reverse the effect of miR-572 on lung cancer cell proliferation inhibition,cycle arrest and apoptosis promotion.Conclusion Down-regulation of miR-572 can inhibit the proliferation and induce apoptosis of lung cancer cells,probably through targeting negative regulation of WWOX.