负载紫杉醇的温敏水凝胶对鼻咽癌的抑制作用

Anti-tumor Effect of Thermosensitive Hydrogel Co-loading Paclitaxel on Nasopharyngeal Cancer

【目的】制备负载紫杉醇(PTX)的壳聚糖/β-甘油磷酸钠(CS/β-GP)温敏水凝胶,表征其理化性质,探讨其对体内外鼻咽癌生长的影响。【方法】在CS/β-GP水凝胶中加入PTX,制备PTX/CS/β-GP水凝胶,扫描电子显微镜观察水凝胶的形态,紫外光谱检测水凝胶的低临界转变温度(LCST)和体外释药性能。体外实验:细胞计数试剂盒(CCK)-8法测定CS/β-GP水凝胶与PTX/CS/β-GP水凝胶对鼻咽癌HNE1细胞活力的影响,4’,6-二脒基-2-苯基吲哚(DAPI)法检测细胞凋亡。体内实验:采用人HNE1细胞皮下注射法建立鼻咽癌移植瘤裸鼠模型,观察PTX和PTX/CS/β-GP水凝胶对移植瘤生长的抑制效果,免疫组织化学法检测移植瘤中Ki-67的表达,末端脱氧核苷酸转移酶介导的dUTP缺口末端标记(TUNEL)法检测移植瘤细胞凋亡。【结果】制备的PTX/CS/β-GP水凝胶呈现规整的三维网络结构、良好的温敏性和药物缓释性能。CS/β-GP水凝胶对HNE1活力无明显抑制作用,PTX/CS/β-GP水凝胶可显著抑制HNE1细胞生长,并促进细胞凋亡。PTX/CS/β-GP水凝胶可显著抑制鼻咽癌移植瘤生长,其抗癌效应优于相同剂量的游离PTX。PTX/CS/β-GP水凝胶可抑制移植瘤组织中Ki-67的阳性表达,并促进TUNEL阳性细胞增多。【结论】CS/β-GP温敏水凝胶具有稳定的理化性质和良好的组织相容性,与游离PTX相比,PTX/CS/β-GP温敏水凝胶具有较低的细胞毒性和更好的抗癌效果,可以作为鼻咽癌治疗的候选药物负载系统。

Objective To prepare paclitaxel(PTX)loaded chitosan/sodiumβ-glycerophosphate(CS/β-GP)thermosensitive hydrogel,its physicochemical properties were characterized and its effects on the growth of nasopharyngeal carcinoma in vitro and in vivo were investigated.Methods PTX/CS/β-GP hydrogel were prepared by adding PTX to CS/β-GP hydrogel.The morphology of the hydrogel was observed by scanning electron microscopy,and the lower critical solution temperature(LCST)and in vitro drug release properties of the hydrogel were detected by UV spectroscopy.In vitro assay:cell counting kit(CCK)-8 assay was used to determine the effect of CS/β-GP hydrogel and PTX/CS/β-GP hydrogel on the viability of nasopharyngeal carcinoma HNE1 cells,and 4’,6-diamidino-2-phenylindole(DAPI)assay was used to detect apoptosis.In vivo experiments:a nasopharyngeal carcinoma xenograft tumor model was established by subcutaneous injection of human HNE1 cells in nude mice to observe the inhibitory effects of PTX and PTX/CS/β-GP hydrogels on xenograft tumor growth,and the expression of Ki-67 in xenograft tumors was detected by immunohistochemistry,and terminaldeoxynucleoitidyl transferase mediated nick end labeling(TUNEL)was used to detect xenograft tumor cell apoptosis.Results The prepared PTX/CS/β-GP hydrogel showed a regular three-dimensional network structure,good thermosensitive status and slow release of drug.CS/β-GP hydrogel did not significantly inhibit HNE1 viability,and PTX/CS/β-GP hydrogel significantly inhibited HNE1 cell growth and promoted apoptosis.The anticancer effect of PTX/CS/β-GP hydrogel was superior to that of free PTX at the same dose.PTX/CS/β-GP hydrogel inhibited positive Ki-67 expression in xenograft tumor tissue and promoted TUNEL-positive cell increase.Conclusion CS/β-GP thermosensitive hydrogel has stable physicochemical properties and good histocompatibility.Compared with free PTX,PTX/CS/β-GP thermosensitive hydrogel has lower cytotoxicity and better anti-cancer effects,and can be used as candidate drug loading systems for the treatment of nasopharyngeal carcinoma.