细胞穿膜肽PFV修饰紫杉醇/青蒿琥酯共载靶向胶束的制备及体外抗肿瘤作用研究(1)
摘 要 目的：制備细胞穿膜肽PFV修饰紫杉醇(PTX)/青蒿琥酯(ART)共载靶向胶束，并考察其体外抗肿瘤活性。方法：按前期优化的工艺，采用薄膜水化法制备PFV修饰PTX/ART共载靶向胶束，并对其进行表征。以空白胶束作为空白对照，采用磺基罗丹明B法评价PTX胶束、ART胶束、PTX/ART胶束以及PFV修饰PTX/ART共载靶向胶束对人胃癌BGC-823细胞的毒性;以香豆素作为荧光探针取代PTX，制成相应的不同胶束，采用流式细胞仪和荧光显微镜测定和观察BGC-823细胞对各胶束的摄取情况和靶向性;并通过Transwell小室法考察PTX胶束、ART胶束、PTX/ART胶束以及PFV修饰PTX/ART共载靶向胶束对BGC-823细胞侵袭的影响。结果：PFV修饰PTX/ART共载靶向胶束的平均粒径为(51.30±3.95) nm，分散系数为0.19±0.01，Zeta电位为(0.21±0.02) mV，PTX、ART的包封率均高于90%，形态呈圆球形。空白胶束对BGC-823细胞无明显毒性，PTX胶束、PTX/ART胶束以及PFV修饰PTX/ART共载靶向胶束对BGC-823细胞的半数抑制浓度分别为(3.09±0.22)、(1.93±0.24)、(1.11±0.15) μmol/L;不同胶束在BGC-823细胞核中的分布数量排序依次为PFV修饰香豆素/ART胶束>香豆素/ART胶束>香豆素胶束>空白对照，抑制细胞侵袭作用的大小依次为PFV修饰PTX/ART共载靶向胶束>PTX/ART胶束>ART胶束>PTX胶束>空白对照。结论：制备的PFV修饰PTX/ART共载靶向胶束符合《中国药典》要求;其对BGC-823细胞具有较强的细胞毒性，可提高药物的靶向性和细胞对药物的摄取能力，并能抑制肿瘤细胞的侵袭和转移。
关键词 细胞穿膜肽;紫杉醇;青蒿琥酯;胶束;人胃癌BGC-823细胞;靶向作用;抗肿瘤作用
ABSTRACT OBJECTIVE： To prepare cell penetrating peptide PFV-modified paclitaxel (PTX)/artesunate (ART) co-loaded targeting micelles， and to investigate in vitro anti-tumor activity. METHODS： According to optimal technology， PFV-modified PTX/ART co-loaded targeting micelles were prepared by membrane hydration method， and were characterized. Using blank micelle as blank control， sulforhodamine B (SRB) method was used to evaluate the toxicity of PTX micelles， ART micelles， PTX/ART micelles and PFV-modified PTX/ART co-loaded targeting micelles to human gastric cancer BGC-823 cells. The coumarin was used as fluorescent probe replacing PTX to prepare corresponding micelles. Then，the uptake of BGC-823 cells to corresponding micelles and targeting effect were observed and determined by flow cytometry and fluorescence microscope. The effects of PTX micelles， ART micelles， PTX/ART micelles and PFV-modified PTX/ART co-loaded targeting micelles on the invasion of BGC-823 cells were investigated by Transwell chamber method. RESULTS： Average particle size of PFV-modified PTX/ART co-loaded targeting micelles was (51.30±3.95) nm; PDI was 0.19±0.01， and Zeta potential was (0.21±0.02) mV. The encapsulation efficiency of PTX and ART were higher than 90%. The shape of micelles were spherical. The blank micelles had no obvious toxicity to BGC-823 cells. The IC50 value of PTX micelles， PTX/ART micelles and PFV-modified PTX/ART co-loaded targeting micelles to BGC-823 cells were (3.09±0.22)， (1.93±0.24)， (1.11±0.15) μmol/L， respectively. The distribution amount of different micelles in BGC-823 cell nucleus in the descending order were PFV-modified coumarin/ART micelles>coumarin/ART micelles>coumarin micelles>blank control. The order of inhibitory effect was PFV-modified PTX/ART co-loaded targeting micelles>PTX/ART micelles>ART micelles>PTX micelles>blank control. CONCLUSIONS： Prepared PFV-modified PTX/ART co-loaded targeting micelles are in line with the quality of Chinese Pharmacopoeia. It shows strong cytotoxicity to BGC-823 cells， can improve the drug targeting and the cell uptake， and inhibit the invasion and metastasis of BGC-823 cells., 百拇医药(王为 李学涛 孔亮 姜爽 罗一夫 王晓波)
关键词 细胞穿膜肽;紫杉醇;青蒿琥酯;胶束;人胃癌BGC-823细胞;靶向作用;抗肿瘤作用
ABSTRACT OBJECTIVE： To prepare cell penetrating peptide PFV-modified paclitaxel (PTX)/artesunate (ART) co-loaded targeting micelles， and to investigate in vitro anti-tumor activity. METHODS： According to optimal technology， PFV-modified PTX/ART co-loaded targeting micelles were prepared by membrane hydration method， and were characterized. Using blank micelle as blank control， sulforhodamine B (SRB) method was used to evaluate the toxicity of PTX micelles， ART micelles， PTX/ART micelles and PFV-modified PTX/ART co-loaded targeting micelles to human gastric cancer BGC-823 cells. The coumarin was used as fluorescent probe replacing PTX to prepare corresponding micelles. Then，the uptake of BGC-823 cells to corresponding micelles and targeting effect were observed and determined by flow cytometry and fluorescence microscope. The effects of PTX micelles， ART micelles， PTX/ART micelles and PFV-modified PTX/ART co-loaded targeting micelles on the invasion of BGC-823 cells were investigated by Transwell chamber method. RESULTS： Average particle size of PFV-modified PTX/ART co-loaded targeting micelles was (51.30±3.95) nm; PDI was 0.19±0.01， and Zeta potential was (0.21±0.02) mV. The encapsulation efficiency of PTX and ART were higher than 90%. The shape of micelles were spherical. The blank micelles had no obvious toxicity to BGC-823 cells. The IC50 value of PTX micelles， PTX/ART micelles and PFV-modified PTX/ART co-loaded targeting micelles to BGC-823 cells were (3.09±0.22)， (1.93±0.24)， (1.11±0.15) μmol/L， respectively. The distribution amount of different micelles in BGC-823 cell nucleus in the descending order were PFV-modified coumarin/ART micelles>coumarin/ART micelles>coumarin micelles>blank control. The order of inhibitory effect was PFV-modified PTX/ART co-loaded targeting micelles>PTX/ART micelles>ART micelles>PTX micelles>blank control. CONCLUSIONS： Prepared PFV-modified PTX/ART co-loaded targeting micelles are in line with the quality of Chinese Pharmacopoeia. It shows strong cytotoxicity to BGC-823 cells， can improve the drug targeting and the cell uptake， and inhibit the invasion and metastasis of BGC-823 cells., 百拇医药(王为 李学涛 孔亮 姜爽 罗一夫 王晓波)