山东医药
山東醫藥
산동의약
SHANDONG MEDICAL JOURNAL
2014年
23期
1-4,110
,共5页
RNA干扰%基因沉默%胰岛素样生长因子1型受体%上皮-间质转化%非小细胞肺癌%表皮生长因子受体-酪氨酸激酶抑制剂%获得性耐药
RNA榦擾%基因沉默%胰島素樣生長因子1型受體%上皮-間質轉化%非小細胞肺癌%錶皮生長因子受體-酪氨痠激酶抑製劑%穫得性耐藥
RNA간우%기인침묵%이도소양생장인자1형수체%상피-간질전화%비소세포폐암%표피생장인자수체-락안산격매억제제%획득성내약
RNA interference%gene silencing%insulin-like growth factor 1 receptor%epithelial-mesenchymal transi-tion%non-small-lung cancer%epidermal growth factor receptor-tyrosine kinase inhibitors%acquired drug resistance
目的：探讨RNA干扰（ RNAi）介导胰岛素样生长因子1受体（ IGF-1R）基因沉默对非小细胞肺癌（NSCLC）细胞PC-9/ZD发生上皮-间质转化（EMT）的影响。方法构建靶向IGF-1R基因的RNAi重组慢病毒，感染NSCLC表皮生长因子受体-酪氨酸激酶抑制剂（ EGFR-TKIs）获得性耐药细胞PC-9/ZD；实时荧光定量PCR法检测IGF-1R表达抑制效应及IGF-1R基因沉默后EMT相关分子的mRNA表达变化，Transwell实验检测细胞侵袭转移能力变化。结果成功构建靶向IGF-1R基因的RNAi重组慢病毒，滴度为1×106 TU/μL，对PC-9/ZD细胞IGF-1R基因的mRNA抑制效率为71．4％。 IGF-1R基因沉默后，PC-9/ZD细胞形态呈现间质向上皮化转变，上皮标志物E-cadherin的mRNA表达量升高（P＜0．05），间质标志物Vimentin的mRNA表达量降低（P＜0．05），且侵袭转移能力减弱（P＜0．05）。结论靶向IGF-1R基因沉默可逆转PC-9/ZD细胞EMT。
目的：探討RNA榦擾（ RNAi）介導胰島素樣生長因子1受體（ IGF-1R）基因沉默對非小細胞肺癌（NSCLC）細胞PC-9/ZD髮生上皮-間質轉化（EMT）的影響。方法構建靶嚮IGF-1R基因的RNAi重組慢病毒，感染NSCLC錶皮生長因子受體-酪氨痠激酶抑製劑（ EGFR-TKIs）穫得性耐藥細胞PC-9/ZD；實時熒光定量PCR法檢測IGF-1R錶達抑製效應及IGF-1R基因沉默後EMT相關分子的mRNA錶達變化，Transwell實驗檢測細胞侵襲轉移能力變化。結果成功構建靶嚮IGF-1R基因的RNAi重組慢病毒，滴度為1×106 TU/μL，對PC-9/ZD細胞IGF-1R基因的mRNA抑製效率為71．4％。 IGF-1R基因沉默後，PC-9/ZD細胞形態呈現間質嚮上皮化轉變，上皮標誌物E-cadherin的mRNA錶達量升高（P＜0．05），間質標誌物Vimentin的mRNA錶達量降低（P＜0．05），且侵襲轉移能力減弱（P＜0．05）。結論靶嚮IGF-1R基因沉默可逆轉PC-9/ZD細胞EMT。
목적：탐토RNA간우（ RNAi）개도이도소양생장인자1수체（ IGF-1R）기인침묵대비소세포폐암（NSCLC）세포PC-9/ZD발생상피-간질전화（EMT）적영향。방법구건파향IGF-1R기인적RNAi중조만병독，감염NSCLC표피생장인자수체-락안산격매억제제（ EGFR-TKIs）획득성내약세포PC-9/ZD；실시형광정량PCR법검측IGF-1R표체억제효응급IGF-1R기인침묵후EMT상관분자적mRNA표체변화，Transwell실험검측세포침습전이능력변화。결과성공구건파향IGF-1R기인적RNAi중조만병독，적도위1×106 TU/μL，대PC-9/ZD세포IGF-1R기인적mRNA억제효솔위71．4％。 IGF-1R기인침묵후，PC-9/ZD세포형태정현간질향상피화전변，상피표지물E-cadherin적mRNA표체량승고（P＜0．05），간질표지물Vimentin적mRNA표체량강저（P＜0．05），차침습전이능력감약（P＜0．05）。결론파향IGF-1R기인침묵가역전PC-9/ZD세포EMT。
Objective To investigate the effect of insulin-like growth factor 1 receptor ( IGF-1R) gene silencing medi-ated by RNA interference ( RNAi) on cell epithelial-mesenchymal transition ( EMT) in non-small-cell lung cancer PC-9/ZD cells.Methods The recombinant lentivirus specific to IGF-1R gene was constructed to be transfected into EGFR-TKIs-resistant non-small-cell lung cancer PC-9/ZD cells.Real-time quantitative RT-PCR was used to detect the inhibition rate of IGF-1R in PC-9/ZD cells;meanwhile, after IGF-1R was inhibited, the mRNA expression of E-cadherin and Vimen-tin was detected , and the invasive and migratory capabilities of cells were determined by Transwell assay .Results The RNAi recombinant lentivirus targeting IGF-1R was successfully constructed .The final titer acquired was 1 ×106 TU/μL, and the inhibition rate of IGF-1R mRNA was 71.4%.After the IGF-1R gene was inhibited, the PC-9/ZD cells displayed epithelial phenotypes, and their capabilities of invasion and migration were significantly decreased (P<0.05).The ex-pression of epithelial cell marker E-cadherin mRNA was increased (P<0.05), accompanied by down-regulation of Vimen-tin mRNA level (P<0.05).Conclusion IGF-1R silencing can reverse cell EMT in non-small-cell lung cancer PC-9/ZD cells.
