中华临床医师杂志（电子版）
中華臨床醫師雜誌（電子版）
중화림상의사잡지（전자판）
CHINESE JOURNAL OF CLINICIANS(ELECTRONIC VERSION)
2014年
22期
3957-3961
,共5页
朱圣韬%李鹏%王青釭%郭水龙%邢洁%程芮%郭庆东%孙秀梅%谷俊朝%张澍田
硃聖韜%李鵬%王青釭%郭水龍%邢潔%程芮%郭慶東%孫秀梅%穀俊朝%張澍田
주골도%리붕%왕청강%곽수룡%형길%정예%곽경동%손수매%곡준조%장주전
环氧化酶2%慢病毒属%癌,鳞状细胞%食管%细胞增殖
環氧化酶2%慢病毒屬%癌,鱗狀細胞%食管%細胞增殖
배양화매2%만병독속%암,린상세포%식관%세포증식
Cyclooxygenase 2%Lentivirus%Carcinoma,squamous cell%Esophagus%Cell proliferation
目的：制备环氧化酶-2（COX-2）的RNA干扰（RNAi）重组慢病毒（lentivirus），观察 COX-2表达沉默对人食管鳞癌（ESCC）细胞增殖的影响。方法将目的基因短发卡 RNA （shRNA）载体与包装载体共转染293T细胞，收集上清液后浓缩纯化、滴度测定、感染ESCC细胞；应用荧光显微镜观察包装效率及感染效率，实时荧光定量PCR及Western blot评价COX-2基因沉默效应，MTS 法检测细胞的增殖。结果成功制备了沉默 COX-2的重组慢病毒，该病毒感染 ESCC细胞后，COX-2 mRNA和蛋白的表达量分别降低了80%和50%左右，细胞的增殖明显减慢。结论慢病毒介导的shRNA干扰能有效沉默ESCC细胞COX-2基因，COX-2表达沉默抑制ESCC细胞增殖。
目的：製備環氧化酶-2（COX-2）的RNA榦擾（RNAi）重組慢病毒（lentivirus），觀察 COX-2錶達沉默對人食管鱗癌（ESCC）細胞增殖的影響。方法將目的基因短髮卡 RNA （shRNA）載體與包裝載體共轉染293T細胞，收集上清液後濃縮純化、滴度測定、感染ESCC細胞；應用熒光顯微鏡觀察包裝效率及感染效率，實時熒光定量PCR及Western blot評價COX-2基因沉默效應，MTS 法檢測細胞的增殖。結果成功製備瞭沉默 COX-2的重組慢病毒，該病毒感染 ESCC細胞後，COX-2 mRNA和蛋白的錶達量分彆降低瞭80%和50%左右，細胞的增殖明顯減慢。結論慢病毒介導的shRNA榦擾能有效沉默ESCC細胞COX-2基因，COX-2錶達沉默抑製ESCC細胞增殖。
목적：제비배양화매-2（COX-2）적RNA간우（RNAi）중조만병독（lentivirus），관찰 COX-2표체침묵대인식관린암（ESCC）세포증식적영향。방법장목적기인단발잡 RNA （shRNA）재체여포장재체공전염293T세포，수집상청액후농축순화、적도측정、감염ESCC세포；응용형광현미경관찰포장효솔급감염효솔，실시형광정량PCR급Western blot평개COX-2기인침묵효응，MTS 법검측세포적증식。결과성공제비료침묵 COX-2적중조만병독，해병독감염 ESCC세포후，COX-2 mRNA화단백적표체량분별강저료80%화50%좌우，세포적증식명현감만。결론만병독개도적shRNA간우능유효침묵ESCC세포COX-2기인，COX-2표체침묵억제ESCC세포증식。
ObjectiveTo construct a recombinant lentivirus for RNA interference (RNAi) of COX-2 gene and observe its effect on esophagealsquamous-cell carcinoma (ESCC) cell proliferation. MethodsThe COX-2 short hairpin RNA (shRNA) and packaging vectors were cotransfected to 293T cells to produce the recombinant lentivirus, which were further ultra-centrifuged, tittered and used to infect ESCC cells. The transfection efficiency and infection efficiency were evaluated with fluorescence microscope. Real-time PCR and Western blot were used to confirm the silencing of COX-2. MTS assay was used to study the effect of COX-2 on cell proliferation.ResultsWe successfully prepared the recombinant lentivirus for RNAi of COX-2 gene. The expression of COX-2 mRNA and protein were effectively silenced and the proliferation of COX-2 silenced cells was significantly inhibited.ConclusionLentivirus mediated shRNA effectively inhibited the expression of COX-2 and the proliferation ability of human ESCC cells.
