中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2013年
3期
505-508
,共4页
孙立臣%潘旭波%周先亭%宋占文%苏长青
孫立臣%潘旭波%週先亭%宋佔文%囌長青
손립신%반욱파%주선정%송점문%소장청
Survivin基因%小发卡RNA%内皮抑素基因%腺病毒%癌,肝细胞
Survivin基因%小髮卡RNA%內皮抑素基因%腺病毒%癌,肝細胞
Survivin기인%소발잡RNA%내피억소기인%선병독%암,간세포
Survivin gene%Small hairpin RNA%Endostatin gene%Adenovirus%Carcinoma,hepatocellular
目的 观察表达Survivin基因小发卡RNA(shRNA)和内皮抑素基因(mE)的双调控双功效的肿瘤特异性溶瘤腺病毒在体内外实验中对肝癌的抗癌活性.方法 以重组的双调控双功效的肿瘤特异性溶瘤腺病毒感染肝癌细胞,Western blot鉴定肝癌细胞基因的蛋白表达,噻唑蓝(MTT)实验检测癌细胞增殖活性；建立肝癌裸鼠移植瘤模型,给予溶瘤腺病毒治疗,观察对移植瘤生长的抑制作用.结果 Western blot实验显示,CNHK500-shRNA-mE病毒增殖能够介导目的基因高效表达,有效抑制Survivin的表达；CNHK500-shRNA-mE感染肝癌细胞后,对肝癌细胞有特异性杀伤作用,在感染强度(MOI)=2时,CNHK.500-shRNA-mE感染组癌细胞存活率下降到50％以下,而对照病毒CNHK500-shRNA、CNHK500-mE、Ad-mE感染组癌细胞存活率均在80％左右；裸鼠SMMC-7721移植瘤模型经病毒治疗后,与对照组比较,CNHK500-shRNA-mE、CNHK500-shRNA、CNHK500-mE、Ad-mE的抑瘤率分别为77.41％ (P＜ 0.01)、59.27％ (P＜ 0.05)、35.96％ (P＜ 0.01)和25.94％ (P＜ 0.05),以CNHK500-shRNA-mE抑瘤作用最强；CNHK500-shRNA-mE介导mE和shRNA高效表达,不但能够抑制间质肿瘤血管新生[对照组血管密度(MVD)=35.6±6.2,CNHK500-shRNA-mE组MVD=11.5 ±3.8,P＜0.01],而且大量诱导癌细胞凋亡[对照组凋亡率为(5.2±1.5)％,CNHK500-shRNA-mE组凋亡率为(26.3±7.5)％,P＜0.01].结论 携带shRNA和mE的双功效溶瘤腺病毒CNHK500-shRNA-mE比任一单功效腺病毒具有更强的肿瘤抑制作用.
目的 觀察錶達Survivin基因小髮卡RNA(shRNA)和內皮抑素基因(mE)的雙調控雙功效的腫瘤特異性溶瘤腺病毒在體內外實驗中對肝癌的抗癌活性.方法 以重組的雙調控雙功效的腫瘤特異性溶瘤腺病毒感染肝癌細胞,Western blot鑒定肝癌細胞基因的蛋白錶達,噻唑藍(MTT)實驗檢測癌細胞增殖活性；建立肝癌裸鼠移植瘤模型,給予溶瘤腺病毒治療,觀察對移植瘤生長的抑製作用.結果 Western blot實驗顯示,CNHK500-shRNA-mE病毒增殖能夠介導目的基因高效錶達,有效抑製Survivin的錶達；CNHK500-shRNA-mE感染肝癌細胞後,對肝癌細胞有特異性殺傷作用,在感染彊度(MOI)=2時,CNHK.500-shRNA-mE感染組癌細胞存活率下降到50％以下,而對照病毒CNHK500-shRNA、CNHK500-mE、Ad-mE感染組癌細胞存活率均在80％左右；裸鼠SMMC-7721移植瘤模型經病毒治療後,與對照組比較,CNHK500-shRNA-mE、CNHK500-shRNA、CNHK500-mE、Ad-mE的抑瘤率分彆為77.41％ (P＜ 0.01)、59.27％ (P＜ 0.05)、35.96％ (P＜ 0.01)和25.94％ (P＜ 0.05),以CNHK500-shRNA-mE抑瘤作用最彊；CNHK500-shRNA-mE介導mE和shRNA高效錶達,不但能夠抑製間質腫瘤血管新生[對照組血管密度(MVD)=35.6±6.2,CNHK500-shRNA-mE組MVD=11.5 ±3.8,P＜0.01],而且大量誘導癌細胞凋亡[對照組凋亡率為(5.2±1.5)％,CNHK500-shRNA-mE組凋亡率為(26.3±7.5)％,P＜0.01].結論 攜帶shRNA和mE的雙功效溶瘤腺病毒CNHK500-shRNA-mE比任一單功效腺病毒具有更彊的腫瘤抑製作用.
목적 관찰표체Survivin기인소발잡RNA(shRNA)화내피억소기인(mE)적쌍조공쌍공효적종류특이성용류선병독재체내외실험중대간암적항암활성.방법 이중조적쌍조공쌍공효적종류특이성용류선병독감염간암세포,Western blot감정간암세포기인적단백표체,새서람(MTT)실험검측암세포증식활성；건립간암라서이식류모형,급여용류선병독치료,관찰대이식류생장적억제작용.결과 Western blot실험현시,CNHK500-shRNA-mE병독증식능구개도목적기인고효표체,유효억제Survivin적표체；CNHK500-shRNA-mE감염간암세포후,대간암세포유특이성살상작용,재감염강도(MOI)=2시,CNHK.500-shRNA-mE감염조암세포존활솔하강도50％이하,이대조병독CNHK500-shRNA、CNHK500-mE、Ad-mE감염조암세포존활솔균재80％좌우；라서SMMC-7721이식류모형경병독치료후,여대조조비교,CNHK500-shRNA-mE、CNHK500-shRNA、CNHK500-mE、Ad-mE적억류솔분별위77.41％ (P＜ 0.01)、59.27％ (P＜ 0.05)、35.96％ (P＜ 0.01)화25.94％ (P＜ 0.05),이CNHK500-shRNA-mE억류작용최강；CNHK500-shRNA-mE개도mE화shRNA고효표체,불단능구억제간질종류혈관신생[대조조혈관밀도(MVD)=35.6±6.2,CNHK500-shRNA-mE조MVD=11.5 ±3.8,P＜0.01],이차대량유도암세포조망[대조조조망솔위(5.2±1.5)％,CNHK500-shRNA-mE조조망솔위(26.3±7.5)％,P＜0.01].결론 휴대shRNA화mE적쌍공효용류선병독CNHK500-shRNA-mE비임일단공효선병독구유경강적종류억제작용.
Objective To investigate the antitumor efficacy of dual-regulated oncolytic adenovirus carrying survivin-small hairpin RNA (shRNA) and mouse endostatin (mE) in hepatocellular carcinoma.Methods The dual-regulated recombinant oncolytic adenovirus was used to infect hepatocellular carcinoma cell lines,and the transgene expression was detected by using Western blotting.The proliferation activity of cancer cells was observed by using methyl thiazol tetrazolium (MTT) assay.The mouse models bearing hepatocellular carcinoma xenografts were established and treated with the oncolytic adenovirus,and the antitumor efficacy was examined.Results The recombinant adenovirus CNHK500-shRNA-mE mediated high expression of transgene selectively in cancer cells,inhibited survivin expression and suppressed cancer cell proliferation.When MOI =2,cell viability was decreased to lower than 50％ in the CNHK500-shRNA-mE group,whereas about 80％ in CNHK-500-shRNA,CNHK500-mE and Ad-mE groups.As compared with the control group,the inhibitory rate of CNHK500-shRNA-mE,CNHK500-shRNA,CNHK500-mE and Ad-mE was77.41％ (P＜0.01),59.27％ (P＜0.05),35.96％ (P＜0.01) and 25.94％ (P＜0.05),respectively,in mouse models bearing hepatocellular carcinoma xenografts,especially in CNHK500-shRNA-mE-treated group.The mE and shRNA mediated by viruses in tumors not only inhibited angiogenesis (MVD =35.6± 6.2 versus 11.5 ± 3.8 in the control and groups,P ＜ 0.01),but also induced cancer cell apoptosis [Apoptosis rate of (5.2 ± 1.5)％ versus (26.3 ±7.5)％ in the control and CNHK500-shRNA-mE groups,P ＜ 0.01].Conclusion The dual-regulated oncolytic adenovirus canying survivin-shRNA and mouse endostatin,CNHK-500-shRNA-mE,can exert more antitumor efficacy than any mono-efficacy adenovirus.
