Noninvasive evaluation of HABP1 expression with Tc-99m-labeled small-interference RNA in ovarian cancer
Noninvasive evaluation of HABP1 expression with Tc-99m-labeled small-interference RNA in ovarian cancer
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使用 Tc-99m 标记的小干扰 RNA 无创评估卵巢癌中 HABP1 的表达
DOI：
10.1080/09553002.2021.1969052
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发表时间：
2021
影响因子：
2.6
通讯作者：
Zhao Changjiu
中科院分区：
文献类型：
--
作者：
Duan Chunyu;Gao Yue;Luan Sha;Guo Shibo;Cao Xueliang;Xu Peng;Fu Peng;Zhao Changjiu
目的卵巢癌是女性最常见的妇科癌症之一，5年生存率低。检测血清透明质酸结合蛋白1(HABP1)水平可为卵巢癌的诊断和治疗提供重要信息。在本研究中，我们设计了一种基于99mTc标记的小干扰RNA(SiRNA)的新型HABP1探针，用于体内无创检测HABP1的表达，从而为HABP1的诊断和治疗提供了新的方法。使用了与人类基因组没有同源性的阴性对照siRNA(NCsiRNA)。用双功能螯合剂联氨烟酰胺(HYNIC)对SiHABP1和NCsiRNA进行99mTc标记。用高效液相色谱法测定了该探针的放化纯度和体外稳定性。Western blotting(WB)和RT-PCR检测其结合活性。高表达HABP1的人卵巢癌细胞系HO-8910用于细胞摄取实验，用伽马计数器测量细胞摄取。Of99mTc-HYNIC-siRNA.Results99mTc-HYNIC-siHABP1在荷瘤小鼠体内分别于注射后1、4、10 h进行显像，并于注射后1、4、6、10 h进行生物分布分析，放化纯度高，体外稳定性好，结合能力和沉默效果与siHABP1相同。SPECT显像显示，在注射99mTC-HYNIC-siHABP1后10 h，肿瘤清晰可见，但在99mTC-HYNIC-NCsiRNA注射后不能清楚地显示肿瘤，提示有特异性结合。结论99mTC-HYNIC-siHABP1是一种无创显示卵巢癌组织中HABP1表达的可行探针。
PurposeOvarian cancer is one of the most common gynecological cancers in women with a low 5-year survival rate. Evaluation of hyaluronic acid-binding protein 1 (HABP1) level can provide important information for the diagnosis and treatment of ovarian cancer. In this study, we designed a novel HABP1 probe based on99mTc-radiolabeled small-interference RNA (siRNA) for detecting HABP1 expression noninvasively in vivo, thereby providing a new method for its diagnosis and treatment.MethodsA specific siHABP1 was selected because of its targetability and silencing effect. A negative control siRNA (NCsiRNA) with no homology with the human genome was used. SiHABP1 and NCsiRNA were radiolabeled with99mTc using the bifunctional chelating agent hydrazinonicotinamide (HYNIC). The radiochemical purity and in vitro stability of the probe were determined by HPLC. The binding activity was measured by western blotting (WB) and RT-PCR. The HABP1-overexpressing human ovarian cancer cell line HO-8910 was used for cell uptake experiments, which were performed with or without transfection and measured with a gamma counter. HO8910-bearing mice were imaged at 1, 4, and 10 h, and biodistribution analysis was performed at 1, 4, 6, and 10 h after injection of99mTc-HYNIC-siRNA.Results99mTc-HYNIC-siHABP1 had high radiochemical purity and good in vitro stability, and showed the same binding capacity and silencing effect as siHABP1. SPECT imaging showed that tumors were clearly visualized at 10 h after injection of99mTc-HYNIC-siHABP1 but not after99mTc-HYNIC-NCsiRNA, implying specific binding. The biodistribution results were consistent with those of SPECT imaging.ConclusionsWe showed that99mTc-HYNIC-siHABP1 is a feasible probe for the noninvasive visualization of HABP1 expression in ovarian cancer.