Xu Yan, Jin Xiangmei, Zhang Jun, Wang Kun, Jin Xiaoyan, Xu Dongyuan, Tian Xizhe, Liu Lan
Center of Morphological Experiment, Yanbian University, Yanji 133002, Jilin, China.
Department of Chemistry, Yanbian University, Yanji 133002, Jilin, China.
ACS Omega. 2020 Oct 6;5(41):26864-26870. doi: 10.1021/acsomega.0c04042. eCollection 2020 Oct 20.
In this study, we designed, formulated, and investigated the potential antitumor activity of a folate receptor (FR)-mediated double-targeted drug delivery system. The system comprised of the FR ligand folic acid (FA), glycine-phenylalanine-leucine-glycine (Gly-Phe-Leu-Gly, GFLG), which can be specifically cleaved by cathepsin B, and the anticancer drug mitomycin C (MMC). The antitumor effect of FA-GFLG-MMC was compared to that of MMC. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay revealed that FA-GFLG-MMC has a significantly higher inhibitory effect on HeLa, SiHa, and PC9 cells (high FR expression) than that on 16HBE and A549 cells (low FR expression). Furthermore, FA-GFLG-MMC inhibited cancer cell proliferation in a dose-dependent manner. Free MMC was toxic to both cancer and normal cells. Apoptosis of the HeLa, SiHa, and PC9 cells was higher than that of the A549 cells; however, the apoptotic effect on 16HBE cells was minimal. Proapoptotic protein bcl-2-associated X-protein (BAX) and antiapoptotic protein BCL-2 play critical roles in cellular defense and apoptotic signal transduction. BAX/BCL-2 ratio is used to determine the intensity of an apoptotic signal and assess whether a cell will survive or undergo apoptosis. BAX and BCL-2 expression in cells treated with 5 μM FA-GFLG-MMC was studied by Western blotting. FA-GFLG-MMC increased the BAX/BCL-2 ratio in HeLa, SiHa, and PC9 cells. The results show that FA-GFLG-MMC can effectively inhibit tumor cell proliferation by inducing apoptosis. Therefore, the system developed can enhance the delivery of anticancer drugs to cancer cells and thereby reduce their toxic effects on normal cells.
在本研究中，我们设计、制备并研究了一种叶酸受体（FR）介导的双靶向药物递送系统的潜在抗肿瘤活性。该系统由FR配体叶酸（FA）、可被组织蛋白酶B特异性切割的甘氨酸-苯丙氨酸-亮氨酸-甘氨酸（Gly-Phe-Leu-Gly，GFLG）和抗癌药物丝裂霉素C（MMC）组成。将FA-GFLG-MMC的抗肿瘤效果与MMC的进行了比较。噻唑蓝比色法显示，FA-GFLG-MMC对HeLa、SiHa和PC9细胞（FR高表达）的抑制作用显著高于对16HBE和A549细胞（FR低表达）的抑制作用。此外，FA-GFLG-MMC以剂量依赖的方式抑制癌细胞增殖。游离MMC对癌细胞和正常细胞均有毒性。HeLa、SiHa和PC9细胞的凋亡率高于A549细胞；然而，对16HBE细胞的凋亡作用最小。促凋亡蛋白bcl-2相关X蛋白（BAX）和抗凋亡蛋白BCL-2在细胞防御和凋亡信号转导中起关键作用。BAX/BCL-2比值用于确定凋亡信号的强度，并评估细胞是存活还是发生凋亡。通过蛋白质免疫印迹法研究了用5μM FA-GFLG-MMC处理的细胞中BAX和BCL-2的表达。FA-GFLG-MMC增加了HeLa、SiHa和PC9细胞中的BAX/BCL-2比值。结果表明，FA-GFLG-MMC可通过诱导凋亡有效抑制肿瘤细胞增殖。因此，所开发的系统可增强抗癌药物向癌细胞的递送，从而降低其对正常细胞的毒性作用。
