nm23-H1小干扰RNA增强紫杉醇脂质体对肺腺癌细胞化疗的敏感性

nm23-H1-siRNA enhances the chemosensitivity to liposome-encapsulated paclitaxel in lung adenocarcinoma cells in vitro

目的探讨抑制nm23-H1基因的表达后，紫杉醇脂质体对人肺腺癌A549细胞化疗敏感性的影响。方法以人肺腺癌A549细胞为研究对象，将其分为nm23-H1-小干扰RNA（siRNA）转染组和未转染组。采用Western blot法检测两组A549细胞中nm23-H1蛋白的表达情况，采用四甲基偶氮唑蓝（M1Tr）法检测紫杉醇脂质体对两组A549细胞的生长抑制率，以流式细胞仪检测A549细胞的凋亡和细胞周期的变化。结果转染nm23-H1-siRNA后，A549细胞中nm23-H1蛋白的表达水平明显降低。紫杉醇脂质体作用48h后，A549细胞的生长抑制率随药物浓度的升高而升高，且转染组细胞的抑制率更高。当紫杉醇脂质体浓度≥5μg/ml时，转染组的抑制效率明显高于未转染组（均P〈0．05）。转染nm23-H1-siRNA后，A549细胞的凋亡率[（65．62±4．36）％]较未转染组明显增加[（43．78±5．56）％，P〈0．05]，S期和G2／M期细胞所占的比例则有所下降[S期：分别为（15．73±3．21）％和（25．56±4．01）％，P〈0．05；G2／M期：分别为（31．91±3．12）％和（39．41±4．21）％，P〈0．05]。结论nm23-H1基因与紫杉醇脂质体的化疗抵抗有关，抑制nm23-H1基因的表达可以增强紫杉醇脂质体的化疗敏感性。

Objective To study the chemosensitivity of lung adenocarcinoma cell line A549 cells to liposome-encapsulated paclitaxel after treatment by nm23-H1-small interference RNA （nm23-H1-siRNA） in vitro. Methods The A549 ceils were divided into two groups： non-transfected group and nm23-H1-siRNA- transfected group. Western blot analysis was used to detect the expression of nm23-H1. MTT and flow cytometry were used to determine the cell mortality rate, apoptosis rate and cell cycle after liposome- encapsulated paclitaxel treatment in both groups. Results The expression of nm23-H1 in A549 cells was significantly decreased after transfection with nm23-H1-siRNA. After treatment for 48 hours with liposome- encapsulated paclitaxel, the cell mortality rate was increased with the increasing concentration of liposome- encapsulated paclitaxel in both groups, but increased higher in the nm23-H1-siRNA-transfected group. When the concentration of liposome-encapsulated paclitaxel was above 5 μg/ml, the cell mortality rate was significantly higher than that in the non-transfected group （P 〈 0.05）. The proportion of apoptotic cells also increased in the nm23-H1-siRNA-transfected group, compared with that of the non-transfected group （t = 3.812, P 〈 0.05 ）, while the proportion of cells at S and G2/M phase decreased after transfection with nm23- H1-siRNA （S phase：t =8. 356,P 〈0.05;G2/M phase：t =7. 256,P 〈0.05）. Conclusions Nm23-H1 is related with the chemoresistance to liposome-encapsulated paclitaxel in lung adenocarcinoma cell line A549 cells. Inhibition of the expression of nm23-H1 by nm23-H1-siRNA can improve the in vitro chemosensitivity of A549 cells to liposome-encapsulated paclitaxel.