In the present study, gold nanoparticles functionalized with anti HER-2 aptamer were designed for effective targeted delivery of dasatinib (DSB) to breast cancer cells. Anti HER-2 aptamer attached to porous or plain gold nanoparticles were compared for dasatinib delivery. Activated drug with succinic anhydride and L-cysteine linker was used for conjugation of DSB to gold nanoparticles. The loading efficiency of the activated drug on plain and porous gold nanoparticles was 52 and 68 %, respectively, which was significantly more than the loading of free DSB in gold nanoparticles (1-2.5 %). The anti HER-2 aptamer was conjugated to porous gold nanoparticles loaded with the activated drug. Various characterization techniques such as FESEM, TEM, AFM, zeta potential and ICP-MS were used to confirm the binding of the drug to gold nanoparticles. 1HNMR and FTIR spectroscopic analyses were employed to examine the structural characteristics of the conjugated drug. These analytical techniques confirmed the successful incorporation of succinyl and thiol groups onto the drug molecule. The amount of aptamer binding to different types of gold nanoparticles was obtained from the intensity of the light emitted from the bands observed in electrophoresis gel and due to the presence of porosity in porous gold nanoparticles, the amount of aptamer conjugation on porous gold nanoparticles increased compared to plain ones. Cell cytotoxicity and cellular uptake were evaluated by MTT assay and TEM in BT-474 and MCF-7 cells. Aptamer-functionalized porous gold nanoparticles containing activated dasatinib showed higher cytotoxicity and cellular uptake than modified DSB-loaded nanoparticles and un-activated DSB. The combination of radiation therapy with the modified dasatinib attached to porous gold nanoparticles and aptamer demonstrated a notable reduction in the IC50 values for both the BT-474 and MCF-7 cell lines. Specifically, the IC50 value for the BT-474 cells decreased from 6.95 μM (for unmodified dasatinib) to 2.57 μM, while for the MCF-7 cells, it decreased from 13.97 μM to 8.57 μM. These findings indicate a significant improvement in the efficacy of the modified dasatinib compared to its unmodified counterpart when used in conjunction with radiation therapy.
A "signal-on" photoelectrochemical sensor for human epidermal growth factor receptor 2 detection based on Y6/CdS organic-inorganic heterojunction.
作者: Jiarong Luo ; Junjun Luo ; Lei Deng ; Minghui Yang ; Xiang Chen
A "signal-on" photoelectrochemical (PEC) immunosensor was successfully constructed for determination of human epidermal growth factor receptor 2 (HER2) based on organic-inorganic heterojunction Y6/CdS as photoactive material. Compared with single organic semiconductor, Y6, Y6/CdS exhibited higher photoelectric conversion efficiency due to the formation of heterojunction. In the presence of HER2, sandwich immune structure was formed between HER2 aptamer and anti-HER2 antibody (Ab) by specific recognition. The polydopamine (PDA) nanoparticles were used for signal amplification to enhance photocurrent intensity as PDA can act as electron donor to eliminate holes and promote electron-hole pairs separation. The developed PEC sensor displayed a wide detection range of 5-1000 pg mL-1 and a low detection limit of 2.2 pg mL-1 for HER2 (S/N = 3). The sensor was successfully used for the detection of HER2 in serum with recoveries between 94.8 and 104% and relative standard deviations (RSDs) in the range of 1.2-4.3%. Furthermore, the designed immunosensor possessed good stability, selectivity, and reproducibility, which can find potential clinical applications for disease diagnosis. A "signal-on" photoelectrochemical sensor was reported for human epidermal growth factor receptor 2 detection based on Y6/CdS organic-inorganic heterojunction.
2020-12-21·International journal of molecular sciences2区 · 生物学
Development of HER2-Specific Aptamer-Drug Conjugate for Breast Cancer Therapy.
2区 · 生物学
作者: Hwa Yeon Jeong ; Hyeri Kim ; Myunghwa Lee ; Jinju Hong ; Joo Han Lee ; Jeonghyeon Kim ; Moon Jung Choi ; Yong Serk Park ; Sung-Chun Kim
In this study, HER2 RNA aptamers were conjugated to mertansine (DM1) and the anti-cancer effectiveness of the conjugate was evaluated in HER2-overexpressing breast cancer models. The conjugate of HER2 aptamer and anticancer drug DM1 (aptamer-drug conjugate, ApDC) was prepared and analyzed using HPLC and mass spectrometry. The cell-binding affinity and cytotoxicity of the conjugate were determined using confocal microscopy and WST-1 assay. The in vivo anti-tumoral efficacy of ApDC was also evaluated in mice carrying BT-474 breast tumors overexpressing HER2. The synthesized HER2-specific RNA aptamers were able to specifically and efficiently bind to HER-positive BT-474 breast cancer cells, but not to HER2-negative MDA-MB-231 breast cancer cells. Also, the HER2-specific ApDC showed strong toxicity to the target cells, BT-474, but not to MDA-MB-231 cells. According to the in vivo analyses drawn from the mouse xenografts of BT-747 tumor, the ApDC was able to more effectively inhibit the tumor growth. Compared to the control group, the mice treated with the ApDC showed a significant reduction of tumor growth. Besides, any significant body weight losses or hepatic toxicities were monitored in the ApDC-treated mice. This research suggests the HER2 aptamer-DM1 conjugate as a target-specific anti-cancer modality and provides experimental evidence supporting its enhanced effectiveness for HER2-overexpressing target tumors. This type of aptamer-conjugated anticancer drug would be utilized as a platform structure for the development of versatile targeted high-performance anticancer drugs by adopting the easy deformability and high affinity of aptamers.