中华核医学与分子影像杂志
中華覈醫學與分子影像雜誌
중화핵의학여분자영상잡지
Chinese Journal of Nuclear Medicine and Molecular Imaging
2014年
5期
379-384
,共6页
赵新明%韩静雅%贾立镯%王娜%张敬勉%王建方%张召奇
趙新明%韓靜雅%賈立鐲%王娜%張敬勉%王建方%張召奇
조신명%한정아%가립탁%왕나%장경면%왕건방%장소기
卵巢肿瘤%寡核苷酸类,反义%受体,表皮生长因子-尿抑胃素%脂质体%小鼠,裸
卵巢腫瘤%寡覈苷痠類,反義%受體,錶皮生長因子-尿抑胃素%脂質體%小鼠,裸
란소종류%과핵감산류,반의%수체,표피생장인자-뇨억위소%지질체%소서,라
Ovarian neoplasms%Oligonucletides,antisense%Receptors,epidemal growth factor-urogastrone%Liposomes%Mice,nude
目的 评价脂质体介导对99Tcm-表皮生长因子受体(EGFR) mRNA反义PNA体外靶细胞摄取、荷瘤裸鼠体内特异显像及生物学分布的影响.方法 利用碱基互补配对原则将带有短肽螯合功能的EGFRmRNA反义PNA与部分互补寡核苷酸杂交,再以配体交换法对反义探针进行99Tcm标记,然后以脂质体包裹反义探针,用HPLC法鉴定标记物的标记率.分析脂质体介导及非脂质体介导99Tcm-EGFR mRNA反义PNA在体外人卵巢癌SKOV3细胞中的摄取率及滞留率的差别,同时分析两者在荷SKOV3卵巢癌裸鼠模型体内生物学分布及显像情况的差异.数据分析采用两样本t检验(或t'检验)及Wilcoxon秩和检验.结果 脂质体介导及非脂质体介导99Tcm-EGFR mRNA反义PNA6h内标记率均在95％以上.两者在注射后1、2、4、6、12及24 h后的细胞摄取率分别为(28.90±1.12)％、(32.76±1.20)％、(38.20±3.11)％、(41.23±1.60)％、(46.63±1.55)％和(46.78±2.14)％,(3.51±0.39)％、(3.90±0.40)％、(4.69±0.18)％、(5.91±0.26)％、(5.30±0.22)％和(5.39±0.17)％,差异有统计学意义(t'=47.11～58.67,Z=2.80,均P＜0.05),两者滞留率差异亦有统计学意义(t'=7.25～11.55,Z=2.80,均P＜0.05).注射两探针后1h荷瘤裸鼠肿瘤部位均可显像,但脂质体介导使肿瘤显像更为清楚,肿瘤摄取高峰T/NT由3.95上升至5.02,并明显增加注药后各时间点T/NT(t=3.96,t'=12.65～ 14.69,Z=2.83～5.29,均P＜0.05).分子探针主要分布在肿瘤、肾脏及肝脏组织中.肿瘤摄取量随时间逐渐增加,1h时非介导组为(1.49±0.09) ％ID/g,介导后为(2.15±0.21) ％ID/g;6 h时分别为(3.90±0.65) ％ID/g和(5.00±0.10) ％ID/g;脂质体介导后可增加注药后各个时间点的肿瘤/肌肉(t=11.24,t'=3.96～ 11.94,均P＜0.05).结论 脂质体介导可以明显促进99Tcm-EGFR mRNA反义PNA进入细胞,并提高对EGFR高表达肿瘤的显像效果.
目的 評價脂質體介導對99Tcm-錶皮生長因子受體(EGFR) mRNA反義PNA體外靶細胞攝取、荷瘤裸鼠體內特異顯像及生物學分佈的影響.方法 利用堿基互補配對原則將帶有短肽螯閤功能的EGFRmRNA反義PNA與部分互補寡覈苷痠雜交,再以配體交換法對反義探針進行99Tcm標記,然後以脂質體包裹反義探針,用HPLC法鑒定標記物的標記率.分析脂質體介導及非脂質體介導99Tcm-EGFR mRNA反義PNA在體外人卵巢癌SKOV3細胞中的攝取率及滯留率的差彆,同時分析兩者在荷SKOV3卵巢癌裸鼠模型體內生物學分佈及顯像情況的差異.數據分析採用兩樣本t檢驗(或t'檢驗)及Wilcoxon秩和檢驗.結果 脂質體介導及非脂質體介導99Tcm-EGFR mRNA反義PNA6h內標記率均在95％以上.兩者在註射後1、2、4、6、12及24 h後的細胞攝取率分彆為(28.90±1.12)％、(32.76±1.20)％、(38.20±3.11)％、(41.23±1.60)％、(46.63±1.55)％和(46.78±2.14)％,(3.51±0.39)％、(3.90±0.40)％、(4.69±0.18)％、(5.91±0.26)％、(5.30±0.22)％和(5.39±0.17)％,差異有統計學意義(t'=47.11～58.67,Z=2.80,均P＜0.05),兩者滯留率差異亦有統計學意義(t'=7.25～11.55,Z=2.80,均P＜0.05).註射兩探針後1h荷瘤裸鼠腫瘤部位均可顯像,但脂質體介導使腫瘤顯像更為清楚,腫瘤攝取高峰T/NT由3.95上升至5.02,併明顯增加註藥後各時間點T/NT(t=3.96,t'=12.65～ 14.69,Z=2.83～5.29,均P＜0.05).分子探針主要分佈在腫瘤、腎髒及肝髒組織中.腫瘤攝取量隨時間逐漸增加,1h時非介導組為(1.49±0.09) ％ID/g,介導後為(2.15±0.21) ％ID/g;6 h時分彆為(3.90±0.65) ％ID/g和(5.00±0.10) ％ID/g;脂質體介導後可增加註藥後各箇時間點的腫瘤/肌肉(t=11.24,t'=3.96～ 11.94,均P＜0.05).結論 脂質體介導可以明顯促進99Tcm-EGFR mRNA反義PNA進入細胞,併提高對EGFR高錶達腫瘤的顯像效果.
목적 평개지질체개도대99Tcm-표피생장인자수체(EGFR) mRNA반의PNA체외파세포섭취、하류라서체내특이현상급생물학분포적영향.방법 이용감기호보배대원칙장대유단태오합공능적EGFRmRNA반의PNA여부분호보과핵감산잡교,재이배체교환법대반의탐침진행99Tcm표기,연후이지질체포과반의탐침,용HPLC법감정표기물적표기솔.분석지질체개도급비지질체개도99Tcm-EGFR mRNA반의PNA재체외인란소암SKOV3세포중적섭취솔급체류솔적차별,동시분석량자재하SKOV3란소암라서모형체내생물학분포급현상정황적차이.수거분석채용량양본t검험(혹t'검험)급Wilcoxon질화검험.결과 지질체개도급비지질체개도99Tcm-EGFR mRNA반의PNA6h내표기솔균재95％이상.량자재주사후1、2、4、6、12급24 h후적세포섭취솔분별위(28.90±1.12)％、(32.76±1.20)％、(38.20±3.11)％、(41.23±1.60)％、(46.63±1.55)％화(46.78±2.14)％,(3.51±0.39)％、(3.90±0.40)％、(4.69±0.18)％、(5.91±0.26)％、(5.30±0.22)％화(5.39±0.17)％,차이유통계학의의(t'=47.11～58.67,Z=2.80,균P＜0.05),량자체류솔차이역유통계학의의(t'=7.25～11.55,Z=2.80,균P＜0.05).주사량탐침후1h하류라서종류부위균가현상,단지질체개도사종류현상경위청초,종류섭취고봉T/NT유3.95상승지5.02,병명현증가주약후각시간점T/NT(t=3.96,t'=12.65～ 14.69,Z=2.83～5.29,균P＜0.05).분자탐침주요분포재종류、신장급간장조직중.종류섭취량수시간축점증가,1h시비개도조위(1.49±0.09) ％ID/g,개도후위(2.15±0.21) ％ID/g;6 h시분별위(3.90±0.65) ％ID/g화(5.00±0.10) ％ID/g;지질체개도후가증가주약후각개시간점적종류/기육(t=11.24,t'=3.96～ 11.94,균P＜0.05).결론 지질체개도가이명현촉진99Tcm-EGFR mRNA반의PNA진입세포,병제고대EGFR고표체종류적현상효과.
Objective To evaluate the in vitro effect on tumor cell uptake,tumor imaging and in vivo biodistribution of 99Tcm-epidermal growth factor receptor (EGFR) mRNA antisense PNA probe mediated by cationic liposome.Methods The oligonucleotide with sequence complementary to part of the EGFR mRNA antisense PNA was hybridized in an anti-parallel orientation targeted PNA.PNA hybridization complexes were labeled with 99Tcm by ligand exchange.The assembly of lipofectamine and 99Tcm-labeled heteroduplex was achieved by electrostatic interactions,and the radiolabeled purity was determined by reversedphase HPLC (RP-HPLC).The disparities of cell uptake in SKOV3 cells and the differences of biodistribution and molecular imaging in BALB/c nude mice bearing SKOV3 xenografts between lipofectanine-mediated 99Tcm-EGFR mRNA antisense PNA (group 1) and 99Tcm-EGFR mRNA antisense PNA (group 2) were analyzed.Two-sample t (or t') test and Wilcoxon rank sum test were used for statistical analysis.Results The labeling rates of both group 1 and group 2 were more than 95％ within 6 h.The cell uptake at 1,2,4,6,12,24 h after injection was (28.90±1.12)％,(32.76±1.20)％,(38.20±3.11)％,(41.23±1.60)％,(46.63±1.55)％ and (46.78±2.14)％ in group 1,and was (3.51±0.39)％,(3.90±0.40)％,(4.69±0.18)％,(5.91±0.26)％,(5.30±0.22)％ and (5.39±0.17)％ in group 2 respectively (t'=47.11-58.67,Z=2.80,all P＜0.05).The retention ratios showed significant difference between the two groups (t'=7.25-11.55,Z=2.80,all P＜0.05).The SKOV3 tumor could be visualized in both groups at 1 h post injection but much better visualized in group 1.The T/NT ratios were higher in group 1 at all time points (t =3.96,t'=12.65-14.69,Z=2.83-5.29,all P＜0.05).The T/NT ratios at uptake peak were 5.02 and 3.95,respectively.The probe accumulated mainly in tumor,kidneys and liver.Tumor uptake increased with time ((1.49±0.09) ％ID/g and (2.15±0.21) ％ID/g at 1 h,(3.90±0.65) ％ID/g and (5.00±0.10) ％ID/g at 6 h) after lipofectamine treatment.The ratios of tumor to contralateral muscle were also higher in group 1 (t =11.24,t' =3.96-11.94,all P＜0.05).Conclusions Lipofectamine-mediation can significantly improve the intracellular delivery of radionuclide molecular probe.Lipofectamine-mediated 99Tcm-EGFR mRNA antisense PNA can greatly improve imaging contrast and visualization of EGFR-over-expressing tumors.
