RNA干扰沉默E-钙黏蛋白表达对人喉癌Hep-2细胞侵袭功能的影响  

Influencing of hep-2 cell function by RNAi silencing E-cadherin expression

作　　者：田军[1] 李成文 吴桂卿[3] 孙静[4] 陈琪[4] 

机构地区：[1]山东省潍坊市人民医院耳鼻咽喉科,261041 [2]山东省潍坊市昌乐县人民医院耳鼻咽喉科 [3]赣南医学院第一附属医院 [4]潍坊医学院

出　　处：《中华医学杂志》2013年第8期627-630,共4页National Medical Journal of China

摘　　要：目的探讨沉默E-钙黏蛋白（cadherin）基因表达后对人喉癌Hep-2细胞增殖、迁移能力的影响，为喉癌的基因治疗提供理论依据。方法根据Genbank数据库中E-cadherin基因序列，设计、合成3条特异性短发卡RNA（shRNA）；脂质体法将shRNA转染人人喉癌Hep．2细胞；利用实时荧光定量PCR检测shRNA转染Hep-2细胞后对E-cadherin基因的沉默效果；四甲基偶氮唑盐法检测转染后Hep-2细胞体外增殖能力的变化，并计算细胞增殖率（生存率）；用Transwell实验检测转染后Hep-2细胞体外的迁移能力的变化。结果shRNA转染后，干扰组的E-cadherin基因表达比对照组低；干扰组Hep-2细胞的增殖能力较对照组强；Transwell实验中3个干扰组的穿膜细胞数均明显高于阴性对照组，差异均有统计学意义[（262±15）、（288±12）、（292±6）比（74±8）个，均P〈0．01]。结论RNA干扰沉默E-cadherin基因表达后能使Hep-2细胞的增殖、迁移能力明显增强。有望成为喉癌基因治疗的新靶点。Objective To explore the function of human laryngeal carcinoma Hep-2 cell after down- regulating the expression of E-cadherin gene to provide theoretical rationales for gene therapy of laryngeal cancer. Methods According to the GenBank database, 3 pairs of shRNA sequences of E-cadherin gene were designed and synthesized, shRNAs were transfected into the cell line Hep-2 by liposome. Reverse transcription-polymerase chain reaction （RT-PCR） was used to detect the silencing effect of E-cadherin expression. The changed capacity of cell proliferation were detected in vitro by methyl thiazalyl tetrazolium （MTT） assay in the transfected Hep-2 cells and the cell proliferation rate （survival rate） was calculated. And Transwell was used to detect the migratory capacity of Hep-2 cells after siRNA transfection. Results The E-eadherin gene expression of RNAi transfected Hep-2 cells significantly decreased in interference group. And the proliferation of interference group became markedly enhanced. In Transwell test, the migrated cell numbers in interference group were significant higher than those in negative control group （262 ± 15, 288± 12, 292±6 vs 74±8, all P 〈 0. 01 ）. Conclusions RNA interference silencing of E-cadherin gene expression can significantly enhance the proliferation and migratory capacity of Hep-2 cells. And E-cadherin may be considered as one of gene therapy targets for laryngeal cancer.

关 键 词：钙黏着糖蛋白类 RNA干扰 细胞增殖 细胞运动 肿瘤 鳞状细胞 

分 类 号：R739.65[医药卫生—肿瘤]