miR-195调控NEK2对前列腺癌细胞增殖的影响

发布时间：2018-05-29 11:10

  本文选题：miR- + NEK　； 参考：《广东医学》2017年07期

【摘要】：目的研究miR-195调控中心体相关激酶2(NEK2)对前列腺癌细胞增殖的影响。方法 Western blot检测在稳定表达和抑制表达miR-195的LNCaP细胞株中NEK2蛋白的表达水平;构建克隆编码miR-195的序列片段慢病毒载体和表达NEK2的质粒载体,对转染空载体和NEK2,以及同时转染了miR-195模拟物和NEK2、miR-195模拟物和空载体的LNCaP细胞株进行CCK8细胞增殖实验;采用shRNA方法构建干扰NEK2基因表达质粒,NEK2抑制成功和miR-195过表达的LNCaP细胞进行细胞增殖实验,测定si-NC、si-NEK2、si-195的LNCaP细胞的生长速度。结果在过表达miR-195的LNCaP细胞株中NEK2蛋白的表达水平比阴性对照miR-NC的LNCaP细胞株中蛋白表达水平明显降低(P0.01);相反,在miR-195被抑制表达的LNCaP细胞株中NEK2蛋白的表达水平明显增加(P0.01)。细胞增殖实验显示转染NEK2基因的LNCaP细胞在72、96h生长速度显著快于转染空载体的细胞(P0.01);转染miR-195模拟物和NEK2的LNCaP细胞在72、96h生长速度显著慢于单纯转染NEK2的LNCaP细胞(P0.01);si-NEK2的LNCaP细胞在72、96h生长速度明显慢于siNC,且与miR-195过表达的LNCaP细胞相似。结论 miR-195负性调控相关下游基因NEK2的表达,抑制前列腺癌细胞的增殖,初步验证了miRNA-195-NEK2信号轴在前列腺癌中的作用。
[Abstract]:Objective to study the effect of miR-195 on the proliferation of prostate cancer cells. Methods Western blot was used to detect the expression level of NEK2 protein in the LNCaP cell line expressing miR-195 stably and to inhibit the expression of NEK2 protein. The sequence fragment lentivirus vector encoding miR-195 and the plasmid vector expressing NEK2 were constructed. The proliferation of LNCaP cell lines transfected with empty vector and NEK2, as well as miR-195 mimics and NEK2 miR-195 mimics and empty vectors were studied. ShRNA method was used to construct LNCaP cells which interfered with the expression of NEK2 gene and miR-195 overexpression. The growth rate of LNCaP cells of si-NCCnsi-NEK2 si-195 was measured. Results the expression level of NEK2 protein in LNCaP cell line with overexpression of miR-195 was significantly lower than that in LNCaP cell line with negative control miR-NC, but the expression level of NEK2 protein was significantly increased in LNCaP cell line with inhibited miR-195 expression. Cell proliferation test showed that the growth rate of LNCaP cells transfected with NEK2 gene was significantly faster than that of empty vector transfected LNCaP cells at 72 ~ 96 h, and the growth rate of LNCaP cells transfected with miR-195 mimics and NEK2 at 72 ~ 96 h was significantly slower than that of LNCaP cells transfected with NEK2 alone at 72 ~ 96 h. The growth rate of LNCaP cells transfected with miR-195 mimics and NEK2 was significantly slower than that of LNCaP cells transfected with NEK2 alone. The growth rate of SNC cells was significantly slower than that of siNCcells at 72 ~ 96 h, and was similar to that of LNCaP cells overexpressed by miR-195. Conclusion miR-195 negatively regulates the expression of downstream gene NEK2 and inhibits the proliferation of prostate cancer cells, which preliminarily verifies the role of miRNA-195-NEK2 signaling axis in prostate cancer.
【作者单位】： 广州医科大学附属第五医院泌尿外科;广州医科大学附属广州市第一人民医院泌尿外科;广州医科大学附属广州市第一人民医院麻醉科;
【基金】：广东省自然科学基金资助项目(编号:2014A030310501) 广州市医药卫生科技项目(编号:20141A011006)
【分类号】：R737.25

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