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目的:探讨microRNA-150(miR-150)对NK/T细胞淋巴瘤组织和NK-92细胞辐射敏感性的影响。方法:选取2010年1月至2016年1月在南方医科大学珠江医院收治的NK/T细胞淋巴瘤患者36例为研究对象,并收集患者的组织标本。所有患者均接受放疗并采用淋巴瘤国际工作组标准((IWG)评价患者的近期疗效,根据疗效分为完全缓解(CR)组和未完全缓解组(NonCR)。实时荧光定量PCR检测患者肿瘤组织及NK/T细胞淋巴瘤细胞系中miR-150的表达量,向淋巴瘤NK-92细胞转染miR-150 mimics,利用MTT法和集落形成实验分析过表达miR-150对NK-92细胞辐射敏感性的影响,流式细胞术检测转染过表达miR-150对NK-92细胞辐射致凋亡的影响,Western blotting实验检测miR-150对凋亡相关蛋白Caspase3和PARP表达的影响。结果:根据IWG标准,12例患者CR,24例患者为Non-CR;与CR组相比,Non-CR组患者miR-150表达量普遍下调(P<0.05)。与正常对照s CD3-CD56~+NK细胞相比,NK/T细胞淋巴瘤组织(9.10±0.19 vs 4.01±0.22,P<0.01)和5种NK/T细胞淋巴瘤细胞系中miR-150呈明显低表达(P<0.05)。过表达miR-150可明显降低辐射后NK-92细胞的增殖能力(P<0.01)和集落形成能力(P<0.01),明显提高NK-92细胞的辐射增敏比(10 Gy时辐增敏比为5.375),显著促进辐射诱导的NK-92细胞的凋亡[(37.3±1.24)%vs(28.3±2.34)%,P<0.05],可促进激活的Caspase 3和PARP蛋白表达。结论:miR-150在NK/T细胞淋巴瘤组织标本及体外培养细胞系中的表达呈显著低水平,miR-150表达量低的患者放疗后缓解率低;转染miR-150 mimics能够增强辐射对NK-92细胞增殖的抑制作用和促进辐射诱导致凋亡作用,对NK/T细胞淋巴瘤有辐射增敏作用。
Objective: To investigate the effect of microRNA-150 (miR-150) on the radiosensitivity of NK / T cell lymphoma and NK-92 cells. Methods: Thirty-six patients with NK / T cell lymphoma who were admitted to Zhujiang Hospital of Southern Medical University from January 2010 to January 2016 were selected as the research object and the tissue samples of the patients were collected. All patients underwent radiotherapy and the IWG criteria were used to evaluate the short-term efficacy of the treatment and were divided into complete remission (CR) group and non-complete remission group (NonCR) according to the efficacy.Fast real-time fluorescence quantitative PCR was used to detect the tumor tissue And miR-150 in NK / T cell lymphoma cell lines, miR-150 mimics were transfected into NK-92 cells. MTT assay and colony formation assay were used to analyze the effect of miR-150 on NK-92 cells 150 cells were treated with miR-150 to detect the effect of miR-150 on the apoptosis of NK-92 cells.Western blotting was used to detect the effect of miR-150 on the expression of apoptosis related proteins Caspase3 and PARP.Results: According to the IWG criteria, CR was found in 12 patients and Non-CR in 24 patients. The expression of miR-150 in Non-CR group was generally lower than that in CR group (P <0.05) (P <0.01), and NK-T cell lymphoma (9.10 ± 0.19 vs 4.01 ± 0.22, P <0.01) and NK / T cell lymphoma cell lines. Overexpression of miR-150 significantly reduced the proliferation of NK-92 cells (P <0.01) and colony forming ability (P <0.01), and significantly enhanced the activity of NK-92 (Radiation sensitive ratio was 5.375 at 10 Gy), significantly promoted the apoptosis of NK-92 cells ([(37.3 ± 1.24)% vs (28.3 ± 2.34)%, P <0.05] Could promote the expression of activated Caspase 3 and PARP.Conclusion: The expression of miR-150 in NK / T cell lymphoma tissue samples and in vitro cultured cell lines was significantly lower, and the remission rate of patients with low miR-150 expression MiR-150 mimics can enhance the inhibitory effect of radiation on the proliferation of NK-92 cells and promote the radiation-induced apoptosis, and have a radiosensitizing effect on NK / T cell lymphomas.