MiR-34b、靶基因CDK6在骨肉瘤组织中的表达及其临床意义

摘要

[目的]探讨miR-34b、靶基因细胞周期蛋白依赖性激酶6(CDK6)在骨肉瘤中的表达水平及临床意义.[方法]收集2016年3月至2017年3月在本院行开放手术切除术的骨肉瘤组织51例及相应的癌旁正常骨组织(距癌组织边缘至少2cm),采用实时荧光定量PCR(qRT-PCR)检测骨肉瘤组织及癌旁正常组织中miR-34b、CDK6mRNA水平,采用免疫组织化学法(S-P)检测骨肉瘤组织及癌旁正常组织中CDK6蛋白表达水平,分析miR-34b、CDK6表达的相关性及二者与骨肉瘤临床病理特征的相关性.[结果]骨肉瘤组织中miR-34bmRNA表达水平为0.34±0.16,明显低于癌旁组织的1.06±0.13,其差异有统计学意义(Z＝24.942,P＝0.000);骨肉瘤组织中CDK6mRNA表达水平为4.37±0.46,明显高于癌旁组织的1.03±0.12(Z＝50.174,P＝0.000),骨肉瘤组织CDK6蛋白阳性表达率较癌旁组织CDK6蛋白阳性表达率高(78.43％vs17.65％,χ2＝37.744,P＝0.000);miR-34b与CDK6表达呈现明显负性相关(r＝－0.483,P＝0.000);miR-34b、CDK6表达与骨肉瘤患者年龄、性别、肿瘤大小、肿瘤位置无明显相关性(P＞0.05),与Enneking分期、肿瘤转移密切相关(Z＝2.509、3.962,P＝0.016、0.000;2＝6.451、4.217,P＝0.040、0.040).[结论]miR-34b可能通过调控骨肉瘤组织中靶基因CDK6表达,参与骨肉瘤的发生发展.%[Objective]The aim of this study was to investigate the expression and clinical significance of miR-34b and target gene cyclin-dependent kinase 6(CDK6)in osteosarcoma,and to provide a new therapeutic approach for gene therapy of osteosarcoma.[Methods]A total of 51 cases of osteosarcoma patients who underwent surgical resection in our hospital from March 12,2016 to March 12,2017 were selected as the subject of study.There were 51 cases of osteosar-coma and corresponding 51 cases of normal bone tissue(At least 2 cm from the edge of the cancer tissue)were collected.The levels of miR-34b and CDK6 mRNA in osteosarcoma and adjacent normal tissues were detected by qRT-PCR.The expression of CDK6 protein in osteosarcoma and adjacent normal tissues was detected by immunohistochecmistry(S-P method).The relationship between the expression of miR-34b and CDK6 was analyzed,and the correlation between the expression of the two genes and their clinicopathological features of osteosarcom was analyzed as well.[Results] The ex-pression level of miR-34b mRNA in osteosarcoma tissue was 0.34 ± 0.16,which was significantly lower than that in adja-cent tissues(1.06 ± 0.13);the difference was statistically significant(Z ＝ 24.942,P ＝0.000).The expression level of CDK6 mRNA in osteosarcoma tissue was 4.37 ± 0.46,which was significantly higher than that in adjacent tissue(1.03 ± 0.12,Z ＝ 50.174,P＝0.000).The positive expression rate of CDK6 protein in osteosarcoma tissue was 78.43％,which was significantly higher than that in adjacent tissue 17.65％(χ2＝37.744,P ＝0.000).The expression of miR-34b and CDK6 was negatively correlated(r ＝ －0.483,P ＝0.000).The expression of miR-34b and CDK6 were not significantly correlated with age,sex,tumor size and tumor location of osteosarcoma patients(Z ＝ 0.203 、0.384 、0.230 、0.135 ;P ＝0.840 、0.703 、0.819 、0.894.χ2＝0.644 、0.051 、2.403 、0.746 ;P ＝0.422 、0.821 、0.121 、0.689),while itwas closely related to Enneking staging and tumor metastasis(Z ＝ 2.509 、3.962,P ＝0.016 、0.000 ;χ2＝6.451 、4.217,P ＝0.040 、0.040).[Conclusion]miR-34b may be involved in the development of osteosarcoma through regulating the expression of target gene CDK6 gene.