黄连素衍生物(氟[^(19)F]HX-01)体外靶向肝癌的初步研究

Primary study on fluro [^(19)F] berberine derivative for human hepatocellular carcinoma targetting in vitro

氟[^(18)F]标记黄连素衍生物(氟[^(18)F]HX-01)是一种潜在的正电子发射计算机断层显像(PET)肿瘤显像剂,而能量状态不同、无放射性的标准对照品(氟[^(19)F]HX-01)的物理性质及化学性质与其完全相同。本文通过研究氟[^(19)F]HX-01在体外人肝癌细胞和人正常细胞中有无选择性分布的现象,从而为进一步完成活体内氟[^(18)F]HX-01肝癌PET显像奠定基础。本文利用小檗红碱和3-氟丙醇在碱催化作用下,一步反应制备氟[^(19)F]HX01;课题组选取人正常肝细胞(HL-7702)与人肝癌细胞(Hep G2、SMMC-7721)为实验细胞材料,于荧光显微镜下观察[^(19)F]HX-01在细胞内的定位;用细胞增殖活性检测试剂盒(CCK-8)测定氟[^(19)F]HX-01对上述细胞的增殖抑制作用及其对细胞的毒性作用。本文研究结果显示:(1)氟[^(19)F]HX-01与人肝癌细胞Hep G2、SMMC-7721的结合能力明显高于人正常肝细胞HL-7702;(2)氟[^(19)F]HX-01对Hep G2、SMMC-7721以及HL-7702的细胞增殖抑制作用具有剂量依赖效应;(3)氟[^(19)F]HX-01对人正常肝细胞HL-7702的毒性作用较SMMC-7721、Hep G2更低。上述体外研究结果表明:氟[^(19)F]HX-01对人肝癌细胞(Hep G2、SMMC-7721)较人正常肝细胞(HL-7702)具有更高选择性和更高毒性。以此为基础,放射性对照品氟[^(18)F]HX-01有望进一步开发成为潜在肝癌PET显像分子探针。

[19F]HX-01, a Fluorine- 18 labeled berberine derivative, is a potential positron emission tomography （PET） tumor imaging agent, while [19F] HX-01 is a nonradioactive reference substance with different energy state and has the same physical and chemical properties. In order to collect data for further study of [19F]HX-01 PET imaging of hepatocellular carcinoma in vivo, this study compared the uptake of [19F]HX-01 by human hepatocellular carcinoma and normal hepatocytes in vitro. The target compound, [19F]HX-01, was synthesized in one step using berberrubine and 3- fluoropropyl 4-methylbenzenesulfonate. Cellular uptake and localization of [19F] HX-01 were performed by a fluorescence microscope in human hepatocellular carcinoma HepG2, SMMC-7721 and human normal hepatocyte HL-7702. Cellular proliferation inhibition and cell cytotoxicity assay of the [19F]HX-01 were conducted using cell counting kit-8 （CCK-8） on HepG2, SMMC-7721 and HL-7702 cells. Fluorescent microscopy showed that the combining ability of [19F]HX-01 to the carcinoma SMMC-7721 and HepG2 was higher than that to the normal HL-7702. Cellular proliferation inhibition assay demonstrated that [19F] HX-01 leaded to a dose-dependent inhibition on SMMC-7721, HepG2, and HL-7702 proliferation. Cell cytotoxicity assay presented that the cytotoxicity of [19F]HX-01 to SMMC-7721 and HepG2 was obviously higher than that to HL-7702. This in vitro study showed that [19F]HX-01 had a higher selectivity on human hepatocellular carcinoma cells （SMMC-7721, HepG2） but has less toxicity to normal hepatocytes （HL-7702）. This could set up the idea that the radioactive reference substance [18F]HX-01 may be worthy of further development as a potential molecular probe targeting human hepatocellular carcinoma using PET.