中华神经外科杂志
中華神經外科雜誌
중화신경외과잡지
Chinese Journal of Neurosurgery
2008年
4期
259-262
,共4页
于如同%孙卫东%马衍刚%黄冠程
于如同%孫衛東%馬衍剛%黃冠程
우여동%손위동%마연강%황관정
RNA干扰%缺氧诱导因子-1α%人端粒酶逆转录酶%神经胶质瘤%基因
RNA榦擾%缺氧誘導因子-1α%人耑粒酶逆轉錄酶%神經膠質瘤%基因
RNA간우%결양유도인자-1α%인단립매역전록매%신경효질류%기인
RNA interference%Hypoxia-induced factor-1α%Human telomerase reverse transcriptase%Glioma%Gene
目的 研究靶向缺氧诱导因子-1α(HIF-1α)和人端粒酶逆转录酶(hTERT)的短发夹RNA(shRNA),对裸鼠皮下人脑胶质瘤中HIF-1α、hTERT基因表达的抑制作用,及其对肿瘤增殖和细胞凋亡的影响.方法 体外培养人脑胶质瘤U251细胞;建立人脑胶质瘤裸鼠皮下接种模型;将HIF-1α shRNA、hTERT shRNA质粒转染入移植瘤中,观察肿瘤生长情况;HE染色法观察肿瘤组织的病理改变;Western blot检测HIF-1α、hTERT蛋白表达;Annexin V+PI双染流式细胞仪检测凋亡情况.结果 成功建立稳定的裸鼠U251皮下移植瘤模型;治疗5周后HIF-1α shRNA、hTERT shRNA组双基因干扰组肿瘤体积较单基因干扰组及对照组明显减小,抑瘤率为84.2%;病理结果显示双基因干扰组肿瘤生长受到抑制,微血管散在稀疏,大量肿瘤细胞坏死及凋亡;Western blot显示治疗组相应蛋白表达受抑制,双基因干扰组影响二者表达;流式细胞仪检测双基因干扰组肿瘤细胞凋亡率明显高于其他组(P＜0.01).结论 HIF-1α shRNA和hTERT shRNA均可在裸鼠移植瘤体内抑制相应蛋白表达及胶质瘤增殖生长,促进细胞凋亡,而双基因干扰作用更强.
目的 研究靶嚮缺氧誘導因子-1α(HIF-1α)和人耑粒酶逆轉錄酶(hTERT)的短髮夾RNA(shRNA),對裸鼠皮下人腦膠質瘤中HIF-1α、hTERT基因錶達的抑製作用,及其對腫瘤增殖和細胞凋亡的影響.方法 體外培養人腦膠質瘤U251細胞;建立人腦膠質瘤裸鼠皮下接種模型;將HIF-1α shRNA、hTERT shRNA質粒轉染入移植瘤中,觀察腫瘤生長情況;HE染色法觀察腫瘤組織的病理改變;Western blot檢測HIF-1α、hTERT蛋白錶達;Annexin V+PI雙染流式細胞儀檢測凋亡情況.結果 成功建立穩定的裸鼠U251皮下移植瘤模型;治療5週後HIF-1α shRNA、hTERT shRNA組雙基因榦擾組腫瘤體積較單基因榦擾組及對照組明顯減小,抑瘤率為84.2%;病理結果顯示雙基因榦擾組腫瘤生長受到抑製,微血管散在稀疏,大量腫瘤細胞壞死及凋亡;Western blot顯示治療組相應蛋白錶達受抑製,雙基因榦擾組影響二者錶達;流式細胞儀檢測雙基因榦擾組腫瘤細胞凋亡率明顯高于其他組(P＜0.01).結論 HIF-1α shRNA和hTERT shRNA均可在裸鼠移植瘤體內抑製相應蛋白錶達及膠質瘤增殖生長,促進細胞凋亡,而雙基因榦擾作用更彊.
목적 연구파향결양유도인자-1α(HIF-1α)화인단립매역전록매(hTERT)적단발협RNA(shRNA),대라서피하인뇌효질류중HIF-1α、hTERT기인표체적억제작용,급기대종류증식화세포조망적영향.방법 체외배양인뇌효질류U251세포;건립인뇌효질류라서피하접충모형;장HIF-1α shRNA、hTERT shRNA질립전염입이식류중,관찰종류생장정황;HE염색법관찰종류조직적병리개변;Western blot검측HIF-1α、hTERT단백표체;Annexin V+PI쌍염류식세포의검측조망정황.결과 성공건립은정적라서U251피하이식류모형;치료5주후HIF-1α shRNA、hTERT shRNA조쌍기인간우조종류체적교단기인간우조급대조조명현감소,억류솔위84.2%;병리결과현시쌍기인간우조종류생장수도억제,미혈관산재희소,대량종류세포배사급조망;Western blot현시치료조상응단백표체수억제,쌍기인간우조영향이자표체;류식세포의검측쌍기인간우조종류세포조망솔명현고우기타조(P＜0.01).결론 HIF-1α shRNA화hTERT shRNA균가재라서이식류체내억제상응단백표체급효질류증식생장,촉진세포조망,이쌍기인간우작용경강.
Objective To explore the effect of silencing Hypoxia-induced factor-1α(HIF-1α)gene and human telomerase reverse transcriptase(hTERT)gene by short hairpin RNAs(shRNAs)on growth and apoptosis of human gliomas in nude mice.Methods To cuhure human glioma U25 1 cells in vitro and establish models of Human glioma subcutaneous xenograft in nude mice.The volumes of tumors and the inhibition rates were measured five weeks after therapy.Xenograft tumors were observed with HE staining after plasmid therapy.Protein levels of HIF-1α and hTERT were detected by Western blot.Apoptosis of tunlor cells staining with Annexin V and PI were measured by flow cytometer.Resuits The subcutaneous xenograft glioma model of nude mice were constructed steadily.The volumes of tumor in HIF-1α shRNA+hTERT shRNA group was significantly decreased compared with blank plasmid group、PBS group and HIF-1α/hTERT shRNA group.the inhibition rate was 84.2%.Rare natural tumor cells and blood vessel.massive necrosie and apoptotic tumor cells were observed.Protein levels of HIF-1α and/or hTERT were also inhibited.When treated with HIF-1α shRNA and hTERT shRNA.the two protein levels were also inhibited.The apoptosic rate of tumor cells in HIF-1α shRNA+hTERT shRNA group was higher than other groups (P＜0.01).Conclusion HIF-1α shRNA or hTERT shRNA could inhibite the corresponding protein levels and inhibit the Multiplication and growth of gliomas,induce apoptosis of tumor cells.Moreover,the effects of HIF-1α shRNA with hTERT shRNA is better.
