携载盐酸吉西他滨聚乳酸纤维膜可抑制骨肿瘤的生长

摘要

BACKGROUND:Gemcitabine hydrochloride is a water-soluble anticancer drug that induces apoptosis in tumor cells, but it has an excessive release in vivo. OBJECTIVE:To evaluate the effect of poly(lactic acid) (PLA) electrospun fiber membranes carrying gemcitabine hydrochloride on the growth of human osteosarcoma cell lines MG-63. METHODS:PLA electrospun fiber members with (experimental) or without (control) gemcitabine hydrochloride were fabricated and characterized. Two kinds of fiber membranes were immersed in low-glucose DMEM medium, and the supernatants were collected in the two groups at 3, 5, 7 days, respectively. Passage 5 human osteosarcoma cell lines MG-63 were inoculated into 96-well plates containing low-glucose DEME with 15％ fetal bovine serum, and divided into seven groups. Groups 1-3 were cultured in the experimental supernatants of 3, 5, 7 culture days, and groups 4-6 were cultured in the control supernatants of 3, 5, 7 culture days, respectively. The remaining group acted as the negative control with no supernatant. Thereafter, cell counting kit-8 was used to detect cell proliferation, and RT-PCR was used to measure expression of Bcl-2 and Bax at 3 days of culture. RESULTS AND CONCLUSION:(1) No obvious particle was found on the smooth and even surface of the fiber members in the experimental and control groups. There was no significant difference in fiber diameter, contact angle and tensile strength between the two kinds of fiber membranes. (2) The results of cell counting kit-8 showed that compared with the negative control group, the supernatant released from the control group had no effect on the MG-63 proliferation at different time points, while the supernatant released from the experimental group could inhibit the MG-63 proliferation at different time points (P<0.05), and the inhibitory effect became more and more obvious with the prolongation of release time. (3) RT-PCR findings showed that compared with the control group, the supernatant released from the experimental group could increase Bax mRNA expression and decrease Bcl-2 mRNA expression at the same time point. To conclude, the PLA electrospun fiber membranes carrying gemcitabine hydrochloride can sustainably inhibit MG-63 proliferation and promote cell apoptosis.%背景:盐酸吉西他滨是一种水溶性抗癌药,可诱导肿瘤细胞凋亡,但其在体内存在释放过快等缺点.目的:观察载携盐酸吉西他滨聚乳酸静电纺丝纤维膜对人骨肉瘤细胞株MG-63生长的影响.方法:分别制备聚乳酸静电纺丝纤维膜(对照组)与携载盐酸吉西他滨的聚乳酸静电纺丝纤维膜(实验组),并进行表征.将两组纤维膜分别浸泡于DMEM低糖培养基中,3,5,7 d取释放上清液.将第5代骨肉瘤细胞株MG-63接种于96孔板,每孔均加入含体积分数15％胎牛血清的DMEM低糖培养基,分7组培养,其中3组分别加入实验组3,5,7 d的释放上清液,另3组分别加入对照组3,5,7 d的释放上清液,以不加入释放上清液的为阴性对照组,培养3 d后,CCK-8法检测细胞增殖,RT-PCR检测细胞凋亡相关蛋白Bcl-2、Bax表达.结果与结论:①实验组与对照组纤维膜纤维丝平滑均一,表面没有明显的颗粒,两种纤维膜的纤维直径、接触角、拉伸强度比较差异无显著性意义;②CCK-8结果显示,与阴性对照组比较,对照组不同时间点的释放上清液对骨肉瘤细胞株MG-63的增殖无影响;与阴性对照组比较,实验组不同时间点的释放上清液可抑制骨肉瘤细胞株MG-63的增殖(P<0.05),并且随着释放上清液时间的延长,抑制作用越来越明显;③RT-PCR结果表明,与相同时间点对照组释放上清液比较,实验组释放上清液可增加细胞Bax mRNA表达,降低Bcl-2 mRNA表达;④结果表明携载盐酸吉西他滨的聚乳酸静电纺丝纤维膜,可持续抑制人骨肉瘤细胞MG-63的增殖,促进其凋亡.