Apt-siRNA联合多西他赛靶向治疗PSMA阳性前列腺癌及^(99)Tc^(m)-Apt-siRNA的显像监测  

Targeted therapy of Apt-siRNA combining with docetaxel for PSMA-positive prostate cancer and monitoring imaging of ^(99)Tc^(m)-Apt-siRNA

作　　者：教玉颖 李丽[1] 付鹏[1] Jiao Yuying;Li Li;Fu Peng(Department of Nuclear Medicine,1st Hospital of Harbin Medical University,Harbin 150000,China)

机构地区：[1]哈尔滨医科大学附属第一医院核医学科,哈尔滨150000

出　　处：《中华核医学与分子影像杂志》2022年第7期406-411,共6页Chinese Journal of Nuclear Medicine and Molecular Imaging

基　　金：国家自然科学基金(81671714)。

摘　　要：目的前列腺特异膜抗原(PSMA)适配体Apt-A10-3.2可作为前列腺癌早期诊断和靶向治疗的特异性配体。鼠双微体(MDM2)与前列腺癌恶性程度密切相关,且MDM2小干扰RNA(siRNA)可通过RNA干扰机制靶向沉默MDM2目的基因。设计合成PSMA Apt-MDM2 siRNA新型嵌合体,与多西他赛(DTX)联合以探讨PSMA阳性前列腺癌的靶向治疗及^(99)Tc^(m)标记嵌合体显像监测相结合的诊疗新模式。方法PSMA Apt-A10-3.2与MDM2 siRNA通过共价偶联合成嵌合体Apt-siRNA,在PSMA表达阳性的前列腺癌细胞株(22RV1及LNCaP)中,采用Apt-siRNA单独或联合DTX对细胞株进行处理,通过Western blot检测MDM2及凋亡相关蛋白[B淋巴细胞瘤-2基因(Bcl-2)及其相关X蛋白(Bax)、聚ADP核糖聚合酶(PARP)、半胱氨酸蛋白酶3(caspase-3)]表达水平,评价治疗效果。取22RV1荷瘤裸鼠15只,分别予PBS、DTX+Apt(200 pmol)和DTX+Apt(400 pmol)处理,观察肿瘤体积与MDM2水平;行^(99)Tc^(m)-Apt-siRNA SPECT显像,获得肿瘤/肌肉放射性计数(T/M)比值。采用单因素方差分析和Tukey多重检验、线性回归分析等分析数据。结果在22RV1及LNCaP细胞株中,Apt-siRNA明显降低MDM2表达(0.25±0.02,F=183.40,P<0.001;0.56±0.03,F=37.15,P<0.001);Apt-siRNA联合DTX治疗前列腺癌后,Bcl-2表达水平明显降低,Bax、PARP、caspase-3表达水平明显升高。在22RV1荷瘤裸鼠中,Apt-siRNA联合DTX明显抑制肿瘤MDM2蛋白表达(400 pmol:0.59±0.12;F=49.99,P=0.023)及肿瘤体积[400 pmol:(0.22±0.07)cm^(3);F=71.30,P=0.039];SPECT显像示,治疗后T/M比值明显降低(400 pmol:2.07±0.22;F=34.99,P=0.022),与MDM2表达水平有线性回归关系(R^(2)=0.875,P<0.001)。结论Apt-siRNA联合DTX能有效抑制前列腺癌进展,并通过偶联^(99)Tc^(m)实现PSMA阳性前列腺癌可视化靶向诊疗。Objective Apt-A10-3.2(aptamer of prostate specific membrane antigen(PSMA))can be used as a specific ligand for early diagnosis and targeted treatment of prostate cancer.Mouse double minute 2 homolog(MDM2)is closely related to the malignancy of prostate cancer,and MDM2 small interfering RNA(siRNA)can silence MDM2 gene through RNA interference.To design a novel chimera of PSMA Apt-MDM2 siRNA and combine it with docetaxel(DTX)to explore a new diagnosis and treatment model combining targeted therapy of PSMA-positive prostate cancer with ^(99)Tc^(m)-chimera imaging monitoring.Methods Apt-siRNA were obtained by covalent connection of PSMA Apt-A10-3.2 and MDM2 siRNA,which was combined with DTX to treat PSMA-positive prostate cancer cell lines(22RV1 and LNCaP).Cell lines were treated with Apt-siRNA alone or in combination with DTX.The levels of MDM2 and apoptosis-related proteins(B-cell lymphoma-2(Bcl-2),Bcl-2-associated X(Bax),poly ADP-ribose polymerase(PARP),caspase-3)were detected by Western blot,which were used to evaluate the therapeutic effect.Fifteen BALB/c mice bearing 22RV1 xenografts were treated with PBS,DTX+Apt-siRNA(200 pmol)and DTX+Apt-siRNA(400 pmol),respectively.Tumor volume and MDM2 level were observed,and ^(99)Tc^(m)-Apt-siRNA SPECT imaging was performed to obtain the tumor/muscle(T/M)ratio.One-way analysis of variance,Tukey′s test and linear regression analysis were used for data analysis.Results The levels of MDM2 protein were significantly decreased by Apt-siRNA(0.25±0.02,F=183.40,P<0.001;0.56±0.03,F=37.15,P<0.001)in 22RV1 and LNCaP cells.After the treatment of Apt-siRNA+DTX,the levels of Bcl-2 were significantly decreased,and the levels of Bax,PARP and caspase-3 were significantly increased.MDM2 protein level(400 pmol:0.59±0.12;F=49.99,P=0.023)and tumor volume(400 pmol:(0.22±0.07)cm^(3);F=71.30,P=0.039)were significantly inhibited by Apt-siRNA+DTX in mice bearing 22RV1 xenografts.As for ^(99)Tc^(m)-Apt-siRNA SPECT imaging in vivo,T/M ratio of treatment group was significantly decreased(400 pmo

关 键 词：前列腺肿瘤 前列腺特异膜抗原 RNA 小分子干扰 紫杉烷类 锝 放射性核素显像 小鼠 裸 

分 类 号：R737.25[医药卫生—肿瘤]