Chen Jia-ping, Liu Yao, Hu Zhi-bin, Shen Hong-bing
Department of Epidemiology and Biostatistics, Nanjing Medical University, Nanjing, China.
Zhonghua Zhong Liu Za Zhi. 2012 Sep;34(9):664-8. doi: 10.3760/cma.j.issn.0253-3766.2012.09.005.
To investigate the effect of a common polymorphism rs928508(A/G) in flanking region of miR-30c on the expression of pri, pre and mature miR-30c, and discuss the effect of this polymorphism on the maturing process of miR-30c in lung carcinoma.
The pGL3-promoter-miR-30c-A and pGL3-promoter-miR-30c-G luciferase plasmids were created containing A or G allele of miR-30c flanking region. Taqman assay was used to genotype rs928508 polymorphism in 50 lung cancer tissues. RT-PCR was performed to determine the expression of pri-miR-30c, pre-miR-30c, mature miR-30c and miR-30c host gene NFYC in the 50 lung cancer tissues.
The luciferase expression level of the pGL3-promoter-miR-30c-A construct group was not significantly different compared with that in the the pGL3-promoter-miR-30c-G construct group (A549 cells, P = 0.758; 293A cells, P = 0.554; CHO cells, P = 0.175). The results demonstrated that rs928508(A/G) variant had no effect on the transcriptional regulation of pri-miR-30c. In the genotype-phenotype collection analysis of the 50 lung cancer tissues, the expression of pre-miR-30c and mature miR-30c for rs928508 AG/GG genotypes showed significantly lower levels compared with those in the AA genotype (P = 0.009, P = 0.011). However, the expression of pri-miR-30c showed no significant difference between AG/GG genotypes and AA genotype. Similarly, the expression of host NFYC gene was correlated with pri-miR-30c, showed no significant difference between AG/GG genotypes and AA genotype.
The rs928508(A/G) polymorphism in flanking region of miR-30c could influence the processing from pri-miR-30c to mature miR-30c, but does not influence the transcription of pri-miR-30c.
研究miR - 30c侧翼区域常见多态性rs928508（A/G）对pri、pre和成熟miR - 30c表达的影响，并探讨该多态性对肺癌中miR - 30c成熟过程的作用。
构建包含miR - 30c侧翼区域A或G等位基因的pGL3 - promoter - miR - 30c - A和pGL3 - promoter - miR - 30c - G荧光素酶质粒。采用Taqman分析法对50例肺癌组织中的rs928508多态性进行基因分型。运用RT - PCR法检测50例肺癌组织中pri - miR - 30c、pre - miR - 30c、成熟miR - 30c及miR - 30c宿主基因NFYC的表达。
pGL3 - promoter - miR - 30c - A构建体组的荧光素酶表达水平与pGL3 - promoter - miR - 30c - G构建体组相比无显著差异（A549细胞，P = 0.758；293A细胞，P = 0.554；CHO细胞，P = 0.175）。结果表明rs928508（A/G）变异对pri - miR - 30c的转录调控无影响。在50例肺癌组织的基因型 - 表型收集分析中，rs928508 AG/GG基因型的pre - miR - 30c和成熟miR - 30c表达水平与AA基因型相比显著降低（P = 0.009，P = 0.011）。然而，pri - miR - 30c在AG/GG基因型和AA基因型之间的表达无显著差异。同样，宿主NFYC基因的表达与pri - miR - 30c相关，在AG/GG基因型和AA基因型之间无显著差异。
miR - 30c侧翼区域的rs928508（A/G）多态性可影响pri - miR - 30c向成熟miR - 30c的加工过程，但不影响pri - miR - 30c的转录。
