CAJ | 학술논문

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转染入人喉癌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.
목적 탐토침묵E-개점단백(cadherin)기인표체후대인후암Hep-2세포증식、천이능력적영향,위후암적기인치료제공이론의거.방법 근거Genbank수거고중E-cadherin기인서렬,설계、합성3조특이성단발잡RNA (shRNA)；지질체법장shRNA전염입인후암Hep-2세포；이용실시형광정량PCR검측shRNA전염Hep-2세포후대E-cadherin기인적침묵효과；사갑기우담서염법검측전염후Hep-2세포체외증식능력적변화,병계산세포증식솔(생존솔)；용Transwell실험검측전염후Hep-2세포체외적천이능력적변화.결과 shRNA전염후,간우조적E-cadherin기인표체비대조조저；간우조Hep-2세포적증식능력교대조조강；Transwell실험중3개간우조적천막세포수균명현고우음성대조조,차이균유통계학의의[(262±15)、(288±12)、(292±6)비(74 ±8)개,균P＜0.01].결론 RNA간우침묵E-cadherin기인표체후능사Hep-2세포적증식、천이능력명현증강.유망성위후암기인치료적신파점.
Objective To explore the function of human laryngeal carcinoma Hep-2 cell after downregulating 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-cadherin 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.