miR-17-5p及PTEN在弥漫性大B细胞淋巴瘤中的表达及其意义

    Expression and significance of miR-17-5p and PTEN in diffuse large B-cell lymphoma

    • 摘要:
      目的研究弥漫大B细胞淋巴瘤(DLBCL)中微小RNA(miR)-17-5p及磷酸酶张力蛋白同系物(PTEN)表达及其临床意义。
      方法应用荧光实时定量PCR检测82例DLBCL肿瘤组织及30例淋巴结反应性增生组织(对照组)中miR-17-5p及PTEN的表达,统计学分析组间表达差异及两者与临床病理特征之间的关系。根据miR-17-5p序列合成miR-17-5p抑制剂和抑制剂对照,并转染至DLBCL细胞系中,MTT细胞增殖实验及Transwell小室实验观察对肿瘤细胞增殖和侵袭能力的影响。采用Kaplan-Meier生存分析观察不同miR-17-5p及PTEN的表达水平与病人预后之间的差异。
      结果肿瘤组织中miR-17-5p的相对表达量高于正常组织(P < 0.01);肿瘤组织中PTEN的相对表达量低于正常组织(P < 0.01)。Pearson线性相关分析结果表明,肿瘤组织中miR-17-5p与PTEN的表达呈负相关(r=-0.612,P < 0.01)。病人肿瘤原发部位为节内、乳酸脱氢酶(LDH)≤ 245 U/L、临床分期为Ⅰ~Ⅱ期miR-17-5p表达高于节外、LDH>245 U/L、Ⅲ~Ⅳ期,PTEN表达低于节外、LDH>245 U/L、Ⅲ~Ⅳ期,差异均有统计学意义(P < 0.05~P < 0.01)。与转染抑制剂对照相比,转染miR-17-5p抑制剂的SU-DHL-8细胞miR-17-5p表达较低,而PTEN表达较高(P < 0.01)。与转染抑制剂对照相比,转染miR-17-5p抑制剂的SU-DHL-8细胞在第96小时增殖能力均明显下降(P < 0.05);Transwell小室实验中,与转染抑制剂对照组细胞穿膜细胞数比较,转染miR-17-5p抑制剂组明显减少(P < 0.05)。高miR-17-5p组病人3年总生存率低于低miR-17-5p组病人(P < 0.05),低PTEN表达组病人病人3年总生存率低于高PTEN表达组病人(P < 0.05)。
      结论DLBCL肿瘤组织中miR-17-5p表达升高,而PTEN表达降低,两者共同参与DLBCL的发生发展过程,有可能成为新的DLBCL诊断和治疗的肿瘤标志物。

       

      Abstract:
      ObjectiveTo investigate the expression levels of microRNA(miR)-17-5p and phosphotase and tensin homolog(PTEN) in diffuse large B-cell lymphoma(DLBCL), and their clinical significance.
      MethodsThe expression levels of miR-17-5p and PTEN in 82 cases of DLBCL carcinoma tissues and 30 cases of lymph node reactive hyperplasia tissues(control group) were detected using real-time quantitative PCR.The differences of the expression levels of miR-17-5p and PTEN between groups, and their relationship with clinicopathological features were analyzed.According to miR-17-5p sequence, the miR-17-5p inhibitors and inhibitor controls were synthesized, and transfected into the DLBCL cell lines.The MTT cell proliferation assay and Transwell chamber assay were used to observe the effects on the proliferation and invasion ability of tumor cells.The Kaplan-Meier survival analysis was used to analyze the differences between the different expression levels of miR-17-5p and PTEN and prognosis of patients.
      ResultsThe relative expression level of miR-17-5p in tumor tissues was higher than that in normal tissues(P < 0.01).The relative expression level of PTEN in tumor tissue was lower than that in normal tissue(P < 0.01).The results of Pearson linear correlation analysis showed that the expression level of miR-17-5p was negatively correlated with the PTEN expression in tumor tissues(r=-0.612, P < 0.01).The expression level of miR-17-5p in patients with primary tumor site locating intra-ganglia, lactic dehydrogenase(LDH) ≤ 245 U/L and clinical stagesⅠ-Ⅱ were higher than that in patients with primary tumor site locating extra ganglia, LDH>245 U/L and clinical stage Ⅲ-Ⅳ, and the expression level of PTEN in patients with primary tumor site locating intra-ganglia, LDH ≤ 245 U/L and clinical stagesⅠ-Ⅱ was lower than that in extra ganglia, LDH>245 and linical stage Ⅲ-Ⅳ(P < 0.05 to P < 0.01).Compared with the transfection inhibitor control, the expression levels of miR-17-5p and PTEN in SU-DHL 8 cells transfected with miR-17-5p inhibitor were lower and higher, respectively(P < 0.01).Compared with the transfection inhibitor control, the proliferation and invasion ability of DLBCL cells transfected with miR-17-5p inhibitor after 96 h was significantly attenuated(P < 0.05).In Transwell chamber assay, the number of transmembrane cells transfected with miR-17-5p inhibitor was significantly reduced compared with the transfection inhibitor control group(P < 0.05).The 3-year overall survival rate of patients with high miR-17-5p was lower than that of patients with low miR-17-5p(P < 0.05), and that of patients with low PTEN expression was lower than that of patients with high PTEN expression(P < 0.05).
      ConclusionsThe expression level of miR-17-5p increases, while the expression level of PTEN decreases in patients with DLBCL.Both of them are involved in the occurrence and development of DLBCL, and may become a new tumor marker for DLBCL diagnosis and treatment.

       

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