中国实用医药
中國實用醫藥
중국실용의약
CHINA PRACTICAL MEDICAL
2009年
19期
76-78
,共3页
肝癌%survivin%RNA干涉%凋亡%抗肿瘤
肝癌%survivin%RNA榦涉%凋亡%抗腫瘤
간암%survivin%RNA간섭%조망%항종류
Osteosarcoma%Survivin%RNA interference%Apoptosis%Antitumor
目的 探讨针对人 survivin基因的 siRNA对肝癌细胞 HepG2的体内外抑制作用.方法 合成 survivin基因的RNA干涉 (RNA interferance,RNAi)特异性片段 ,构建 survivin特异性的 RNA干涉载体 pSiS,转染 HepG2细胞 , G418筛选稳定转染的细胞系.细胞计数检测细胞生长情况;Western blotting检测细胞中 survivin蛋白表达的变化;通过 Annexin V法染色和流式细胞术检测细胞凋亡;观察裸鼠皮下 HepG2移植瘤的形成.结果 成功构建了 survivin基因 siRNA真核表达载体 pSiS,获得了稳定转染的细胞 HepG2/pSiS.与 HepG2、HepG2/pSi(空质粒对照细胞 )相比 ,HepG2/pSiS细胞生长曲线十分平缓 ,差异有统计学意义 (P< 0. 01);Western blotting显示 ,HepG2/pSiS细胞中 survivin蛋白表达减少 ,细胞凋亡率增加 (P< 0. 01).HepG2/pSiS移植瘤形成明显受到抑制.结论 Survivin特异性 siRNA可明显促进肝癌细胞 HepG2的凋亡, 抑制该肿瘤细胞体外生长和体内移植成瘤.
目的 探討針對人 survivin基因的 siRNA對肝癌細胞 HepG2的體內外抑製作用.方法 閤成 survivin基因的RNA榦涉 (RNA interferance,RNAi)特異性片段 ,構建 survivin特異性的 RNA榦涉載體 pSiS,轉染 HepG2細胞 , G418篩選穩定轉染的細胞繫.細胞計數檢測細胞生長情況;Western blotting檢測細胞中 survivin蛋白錶達的變化;通過 Annexin V法染色和流式細胞術檢測細胞凋亡;觀察裸鼠皮下 HepG2移植瘤的形成.結果 成功構建瞭 survivin基因 siRNA真覈錶達載體 pSiS,穫得瞭穩定轉染的細胞 HepG2/pSiS.與 HepG2、HepG2/pSi(空質粒對照細胞 )相比 ,HepG2/pSiS細胞生長麯線十分平緩 ,差異有統計學意義 (P< 0. 01);Western blotting顯示 ,HepG2/pSiS細胞中 survivin蛋白錶達減少 ,細胞凋亡率增加 (P< 0. 01).HepG2/pSiS移植瘤形成明顯受到抑製.結論 Survivin特異性 siRNA可明顯促進肝癌細胞 HepG2的凋亡, 抑製該腫瘤細胞體外生長和體內移植成瘤.
목적 탐토침대인 survivin기인적 siRNA대간암세포 HepG2적체내외억제작용.방법 합성 survivin기인적RNA간섭 (RNA interferance,RNAi)특이성편단 ,구건 survivin특이성적 RNA간섭재체 pSiS,전염 HepG2세포 , G418사선은정전염적세포계.세포계수검측세포생장정황;Western blotting검측세포중 survivin단백표체적변화;통과 Annexin V법염색화류식세포술검측세포조망;관찰라서피하 HepG2이식류적형성.결과 성공구건료 survivin기인 siRNA진핵표체재체 pSiS,획득료은정전염적세포 HepG2/pSiS.여 HepG2、HepG2/pSi(공질립대조세포 )상비 ,HepG2/pSiS세포생장곡선십분평완 ,차이유통계학의의 (P< 0. 01);Western blotting현시 ,HepG2/pSiS세포중 survivin단백표체감소 ,세포조망솔증가 (P< 0. 01).HepG2/pSiS이식류형성명현수도억제.결론 Survivin특이성 siRNA가명현촉진간암세포 HepG2적조망, 억제해종류세포체외생장화체내이식성류.
Objective To explore the inhibitory effect of survivin-targeted siRNA on liver cancer cell line HepG2 in vivo and in vitro. Methods According to the survivin cDNA sequence, the specific RNA interference (RNAi) fragmentswere designed and synthesized, which were then cloned into pSilencer 3. 02H1 neo plasmid vector to construct pSiS vector. HepG2 cellswere transfected with RNAi vectors and negative control vector separately;the stably transfected cell strains were selected by G418;and cell growth was assessed by cell counting. Expression of survivin protein was investigated byWestern blotting. Apoptosis analysiswas examined byAnnexinⅤmethod. Tumorigenesis assaywas conducted in nudemice. Results The specific survivin-targeted siRNA eukaryotic vectorpSiSwas successfully constructed, and transfectants were obtained. Compared with blank vector transfected cells and untransfected cells, HepG2/pSiS cells had a slow growth (P<0.01). Expression of survivin protein was significantly inhibited inHepG2/pSiS cells, which had an increased apoptotic rate (P<0.01). The tumorigenesis of transplanted HepG2/pSiS was greatly inhibited. Conclusion Survivin-targeted siRNA can promote the apoptosis of HepG2 cells, and inhibit the in vitro growth and in vivo tumor formation in nude mice.
