摘要
目的探讨纳米羟基磷灰石（nHAP）介导靶向端粒酶逆转录酶（hTERT）的RNA干扰对人肺癌A549细胞的影响。方法采用均相沉淀法制备nHAP。分为nHAP—PLL组、脂质体组、nHAP组及对照组，分别配制转染复合物并转染人肺癌A549细胞。MTT法测定细胞生长活力；Western Blot检测hTERT蛋白表达，流式细胞仪检测细胞凋亡率。结果nHAP-PLL组、脂质体组与nHAP组的A549细胞增殖受到抑制，较对照组差异有统计学意义（P〈0．05）；nHAP—PLL组抑制细胞增殖较脂质体组及nHAP组明显，差异均有统计学意义（P〈0．05）。各组的hTERT蛋白表达的变化趋势与人肺癌A549细胞的增殖情况一致。流式细胞仪检测nHAP—PLL组、脂质体组及nHAP组均有不同程度的细胞凋亡，凋亡率与对照组差异有统计学意义，P〈0．05。结论nHAP本身能诱导人肺癌A549细胞凋亡，经PLL修饰后能有效地结合保护DNA并介导人肺癌A549细胞的基因转染，介导靶向hTERT的RNA干扰可有效抑制人肺癌A549细胞增殖。
Objective To investigate the effect of hydroxyapatite nanoparticles （nHAP） mediated human telomerase reverse transcriptase （hTERT） RNA interference of A549 human lung cancer cells in vitro. Methods The nHAP were synthesized by the homogeneous precipitation method. The structure of the nanoparticles was observed under transmission electron microscope. The nHAP were prepared using uhrasonication and Na2CO3 and modified with poly-L-lysine （PLL） at pH 7.4. The transfection of pGenesil-hTERT into A549 was divided into four groups as follows ： nHAP-PLL group mediated by hydroxyapatite nanoparticles modified with poly-L-lysine （nHAP-PLL）, liposome group mediated by Lipofectamine, nHAP group mediated by hydroxyapatite nanoparticles and control group. The growth ability of cells was assayed with methyl thiasolyl tetrazolium method. The expression level of hTERT protein was examined by Western blotting. Flow cytometry was used to detect the apoptosis ratio of A549 cells line. Results Under transmission electron microscope, the synthesized product presented needle-like and well dispersed particles with evenly distributed sizes of （15 -20） nm × （60-80） nm. The proliferation of A549 cells of nHAP-PLL group, liposome group and nHAP group were obviously inhibited as compared with the control group （ P 〈 0.05 ）. The inhibition rate of nHAP-PLL group was more than the other groups. There was a significant difference inhibition rate between the nHAP-PLL group compared with the liposome group and nHAP group （ P 〈 0.05 ）. The level of hTERT protein had a similar varietal tendency with the result of proliferation of each group. Flow cytometry showed the apoptosis ratio of nHAP-PLL group, liposome group, nHAP group and control group was （28.1 ±1.4） %, （ 19.2 ±1.3 ） %, （ 10.9 ±1.2） % and （0.3 ±0.2 ） %, respectively. There was a significant difference in apoptosis ratio between the nHAP-PLL group, liposome group and nHAP group compared with control group（P 〈 0.05 ）. Conclu
出处
《中华胸心血管外科杂志》
CSCD
北大核心
2010年第1期41-44,共4页
Chinese Journal of Thoracic and Cardiovascular Surgery
