PLLA-PTX微粒的缓释及对人卵巢癌SKOV3细胞的抑瘤作用

Controlled Release of Paclitaxel from Microparticles Containing PLLA and Its Anti-tumor Activity on Human Ovarian Carcinoma Cell Line SKOV3

目的证明聚左旋乳酸-紫杉醇(PLLA-PTX)微粒影响SKOV3细胞增殖和凋亡的有效性;探讨生物降级高分子材料聚左旋乳酸(PLLA)对所担载抗肿瘤药物紫杉醇(PTX)的缓释作用。方法实验分为PLLA-PTX微粒组、PTX组和空白对照组。MTT比色法检测细胞的增殖情况;倒置显微镜观察细胞形态学变化;FCM技术进行细胞凋亡及周期分析;TUNEL检测、定位凋亡细胞。结果PLLA-PTX微粒能有效的抑制SKOV3细胞的增殖,具剂量和时间依赖性。其抑瘤作用明显优于裸露的PTX,随时间的延长,优势不断增大。PLLA-PTX微粒的最佳实验浓度为0.05μmol/L。倒置显微镜下,PLLA-PTX微粒组和PTX组培养液中漂浮大量体积变圆的细胞和颗粒状无定形碎片,随时间的延长,PLLA-PTX微粒组更明显。PLLA-PTX微粒组和PTX组肿瘤细胞凋亡率均高于对照组(P<0.05)。随时间的延长,PLLA-PTX微粒提高细胞凋亡率、增加G2/M期细胞比例的优势大于PTX组。TUNEL可以准确定位PLLA-PTX微粒作用后正在凋亡的细胞,同时出现了两种独特细胞:病理性核分裂象细胞和多微核细胞。结论PLLA-PTX微粒具有较强的细胞毒作用及明显的缓释作用,可以维持较长时间的有效浓度,持续抑制SKOV3细胞的增殖。

Objective To prove the PLLA-PTX＇ efficacy on growth, apoptosis of ovarian cancer cell line SKOV3, and to study the controlled release roles of PLLA for PTX. Methods Cultured cells of human ovarian Carcinoma cell Line SKOV3 were treated with PLLA-PTX microparticles and PTX only separately, untreated cells as the control. The proliferation of SKOV3 cells were determined by MTT assay with the morphologic change observed under inverted phase contrast microscope, the apoptosis of cell were demonstrated by FCM and in situ TUNEL technique. Results The anti-tumor activity of PLLA-PTX microparticles was stronger than PTX alone. A time-dependent and dose-dependent growth inhibition was abserved. At 0.05 μmol/L drug concentration, SKOV3 cell viability experiment demonstrated that the drug formulated in the microparticles was more effective than that formulated in PTX. PLLA-PTX microparticles can increase G2/M period percentage, interrupt the cell cycle proceeding, and inhibit the tumor cells＇growth through cell apoptosis. Positive staining for the presence of apoptosis were obtained in SKOV3 cells with PLLA-PTX microparticles. Conclusions PLLA-PTX microparticles have strong anti-tumor activity and obviously controlled released effectiveness. It shows longer and stronger controlled antitumor activity than paclitaxel alone.