紫杉醇/NLS-KALA-SA核定位纳米粒对肺腺癌A549细胞株的抑制作用

Inhibition effects of paclitaxel/NLS-KALA-SA nanoparticles on A549 cell line in vitro

目的观察多肽自组装载体载紫杉醇纳米粒(NKSP)及紫杉醇单药对肺腺癌A549细胞株的体外抑制作用及可能机制。方法用噻唑蓝(MTT)法分别检测不同浓度组NKSP(20、40、80、100μg/L)、紫杉醇单药(20、40、80、100μg/L)作用24、48及72 h对A549细胞增殖的影响。流式细胞仪检测不加任何药物处理(A)组、加入80μg/L的多肽自组装纳米粒(NKS)培养液(B)组、加入80μg/L紫杉醇单药(C)组、加入含80μg/L NKSP(D)组作用A549细胞48 h及72 h时的细胞凋亡率。Western blot检测A、B、C、D组作用48 h及72 h细胞凋亡相关蛋白bax、caspase-3的表达。结果紫杉醇单药、NKSP均能抑制A549的增殖,紫杉醇单药各组48和72 h时、NKSP单药各组在72 h时随浓度递增抑制率也呈递增趋势(均P<0.05)。48 h时D组促A549凋亡作用低于C组(P<0.05),72 h强于C组(P<0.05),且药物作用48 h时D组的bax、caspase-3的表达低于C组,而72 h时高于C组(P<0.05)。结论 NKS包裹紫杉醇后可促使载体内的紫杉醇缓慢释放,与紫杉醇单药相比,可降低细胞毒性,延长抗肿瘤作用时间。

Objective To observe NLS-KALA-SA-PTX （NKSP） for lung adenocarcinoma cell line A549 in vitro with paclitaxel monotherapy, and the mechanism thereof. Methods MTT assay was used to detect A549 cell proliferation influenced by different concentrations of NKSP （20, 40, 80, 100 μg/L） and paclitaxel monotherapy （20, 40, 80, 100 μg/L） for 24 h, 48 h and 72 h. Subsequent experiments were divided into four groups, namely, group A （without any drug treatment）, group B （added polypeptide 80 μg/L of self-assembled nanoparticles, NKS）, group C （80 μg/L paclitaxel monotherapy） and group D （80 μg/L NKSP）. Flow cytometry was used to detect the cell apoptotic rates after 48 h and 72 h treatment in four groups. Western blot assay was used to analyse the protein expressions of bax and easpase-3 after 48 h and 72 h treatment in four groups. Results Both paclitaxel monotherapy and NKSP can inhibit the proliferation of A549 ceils. The inhibitory rates of paclitaxel monotherapy group at 48 h and 72 h and NKSP group at 72 h showed an increasing trend in a dose-depen- dent manner （P 〈 0.05）. After treatment for 48 hours, the apoptotic rate was significantly higher in D group than that of C group （P 〈 0.05）. But the apoptotic rate at 72 h was lower in D group than that of C group （P 〈 0.05）. The protein expressions of bax and caspase-3 at 48 h were significantl lower in D group than those of C group, which were higher at 72 h in D group than those of C group （P 〈 0.05）. Conclusion Compared to paclitaxel monotherapy group, NKS promotes slow release of pa- clitaxol, which reduces the cytotoxicity and extends the antitumor effects.