摘要
目的研究miR-33b与高迁移率族蛋白A2(HMGA2)在食管鳞状细胞癌(ESCC)中的表达及意义。方法在ESCC组织及癌旁组织中采用茎环结构的逆转录实时定量PCR检测miR-33b的表达情况,实时荧光定量PCR检测HMGA2 mRNA的水平,免疫组织化学SP法检测HMGA2蛋白表达;将miR-33b模拟物、miR-33b抑制物、对照转染入TE-1细胞和Eca-109细胞;采用茎环结构的逆转录实时定量PCR法检测各组miR-33b的水平;实时荧光定量PCR、Western blot法分别检测各组HMGA2mRNA和蛋白的水平。结果在40例ESCC组织中,miR-33 b mRNA的表达量显著低于癌旁正常组织;HMGA2 mRNA的表达量显著高于癌旁正常组织;HMGA2蛋白在ESCC组织中的阳性率显著高于癌旁组织;HMGA2蛋白阳性表达组的miR-33b的表达量低于HMGA2阴性组;相关性分析显示miR-33b和HMGA2的mRNA在ESCC组织中的表达呈负相关。在转染miR-33b模拟物、miR-33b抑制物转染的食管癌细胞中,转染各组间HMGA2 mRNA水平无显著差异;在miR-33b过表达的细胞中HMGA2蛋白水平明显降低。结论 ESCC组织中miR-33b表达下调,HMGA2表达上调;增加食管癌细胞中miR-33b的表达能抑制HMGA2的蛋白表达水平,但HMGA2 mRNA水平不受影响,提示miR-33 b可能在转录后水平调控HMGA2蛋白的表达。
Objective To investigate the expression and significance of miR-33b and high mobility group AT-hook 2 （HMGA2） in esophageal squamous cell carcinoma （ESCC）. Methods Real-time quantitative PCR （qRT-PCR） was used to detect the expression levels of miR-33b and HMGA2 mRNA from the ESCC and adjacent tissues. performed to test the protein level of HMGA2. After the miR-33b mimics, inhibitor and controls were separately transfected into TE-1 and Eca-109 cells, the expression levels of miR-33b and HMGA2 in each group were again determined by qRT-PCR and Western blotting. Results The expression level of miR-33b from the ESCC tissues of 40 cases were significantly lower than that from the adjacent normal tissues. However, the expression level of HMGA2 protein from the ESCC tissues were significantly higher than that from the adjacent normal tissues. The expression level of miR-33b in the HMGA2 positive group was lower than that in the HMGA2 negative group. The correlation analysis showed that the expression of miR-33b was negatively correlated with the mRNA expression of HMGA2 in the ESCC tissues. There were no significant difference in the mRNA expression of HMGA2 among the cells transfected separately with miR-33b mimics, inhibitor and controls, but Western blotting indicated that the protein expression of HMGA2 decreased significantly in the miR-33b over-expressed cells. Conclusion The expression of miR-33b decreases and HMGA2 increases in the ESCC tissues. The over-expression of miR-33b could suppress the protein expression of HMGA2 in the esophageal cancer cells, but the mRNA expression of HMGA2 would not be affected,which suggests that miR-33b might regulate the protein expression of HMGA2 in the post-transcriptional level.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2016年第9期1238-1242,1247,共6页
Chinese Journal of Cellular and Molecular Immunology
基金
常州市科技计划项目(CJ20140002)
