微小RNA-374a靶向脾酪氨酸激酶对卵巢癌细胞放射敏感性及迁移、侵袭的影响

MiR-374a affects radiosensitivity and migration and invasion of ovarian cancer cells by targeting Syk

目的探讨微小RNA-374a(miR-374a)靶向脾酪氨酸激酶(Syk)对卵巢癌细胞迁移、侵袭的影响及其与放射敏感性的关系。方法选取2017年2月至2018年1月宜宾市第二人民医院接受手术切除治疗的卵巢癌病人63例,术中切除卵巢癌组织及癌旁组织,以癌旁组织为对照,采用实时荧光定量逆转录聚合酶链反应(qRT-PCR)检测卵巢癌组织及癌旁组织中miR-374a的表达水平;体外培养卵巢癌A2780细胞,采用随机数字表法分成anti-miR-NC组、anti-miR-374a组、anti-miR-374a+si-NC组、anti-miR-374a+si-Syk组。采用MTT法检测各组细胞增殖活性;克隆形成实验检测各组细胞存活分数及放射敏感性;Tran-swell实验检测各组细胞迁移及侵袭数;miR-374a与Syk的靶向调控作用通过双荧光素酶报告基因检测验证。结果与癌旁组织比较,卵巢癌组织中miR-374a的表达水平升高[(0.28±0.03)比(0.92±0.09)](P<0.05);与anti-miR-NC组比较,anti-miR-374a组细胞存活分数降低[(0.37±0.08)比(0.07±0.02)](P<0.05),增敏比(1.816)增加,迁移[(139±14)个比(66±7)个]及侵袭细胞数[(127±13)个比(58±6)个]减少(P<0.05),细胞增殖能力降低[24 h(0.65±0.06)比(0.30±0.03);48 h(1.04±0.10)比(0.45±0.04);72 h(1.63±0.16)比(0.60±0.06)](P<0.05);miR-374a可靶向结合Syk,并可负向调控Syk的表达与活性;抑制Syk表达可减弱miR-374a抑制对细胞生物学行为及放射敏感性的影响。结论miR-374a低表达可通过上调Syk表达抑制卵巢癌细胞增殖、迁移及侵袭,提高细胞对放射治疗的敏感性。

Objective To investigate the effect of microRNA-374a(miR-374a)targeting spleen tyrosine kinase(Syk)on migration and invasion of ovarian cancer cells and its relationship with radiosensitivity.Methods A total of 63 patients with ovarian cancer who underwent surgical resection from February 2017 to January 2018 in Yibin Second People's Hospital were selected.The ovarian cancer tissues and adjacent tissues were removed during the operation.The adjacent tissues were used as controls,qRT-PCR was used to de-tect the expression level of miR-374a in ovarian cancer tissues and adjacent tissues.Ovarian cancer cells A2780 were cultured in vitro and divided into anti-miR-NC group,anti-miR-374a group,anti-miR-374a+si-NC group,anti-miR-374a+si-Syk group by random num-ber table method.The proliferation of each group was detected by MTT assay.The colony formation assay was used to detect the surviv-al fraction and radiosensitivity of each group.Cell migration and invasion numbers was calculated using Transwell test.The targeted regulation of miR-374a and Syk was determined using dual luciferase reporter assays.Results Compared with adjacent tissues,the ex-pression level of miR-374a in ovarian cancer tissues was increased[(0.28±0.03)vs.(0.92±0.09)](P<0.05).Compared with anti-miR-NC group,the survival fraction of anti-miR-374a group decreased[(0.37±0.08)vs.(0.07±0.02)](P<0.05),the sensitization ratio(SER)(1.816)increased,the number of migration[(139±14.18)vs.(66±6.83)]and invasion cells[(127±13.21)vs.(58±5.86)]decreased(P<0.05),and the cell proliferation ability decreased significantly[24 h(0.65±0.06)vs.(0.30±0.03);48 h(1.04±0.10)vs.(0.45±0.04);72 h(1.63±0.16)vs.(0.60±0.06)](P<0.05).miR-374a could target Syk and negatively regulated the expression and activity of Syk.Inhibiting Syk expression reversed the effect of miR-374a inhibition on cell biological behaviour and radiosensitivity.Conclusion Low expres-sion of miR-374a inhibits cell proliferation,migration and invasion in ovarian cancer cells through up-regulating Syk,and increases the sensitivity of cells to radiotherapy.