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目的:探讨新型纳米材料聚酰胺-胺型树枝状高聚合物(polyamidoamine dendrimer,PAMAM)介导NK4基因对乳腺癌MDA-MB-231、MCF-7细胞生长的抑制作用及对MDA-MB-231细胞移植瘤裸鼠模型的治疗作用。方法:制备PAMAM-NK4纳米复合物颗粒,纳米复合物和空载体PAMAM分别转染MDA-MB-231和MCF-7细胞作为实验组和对照组。MTT实验、Western blotting和流式细胞术分别检测转染PAMAM-NK4对细胞增殖、NK4蛋白表达和细胞凋亡的影响。40只雌性裸鼠经皮下注射建立MDA-MB-231细胞移植瘤裸鼠模型,随机分成4组,每组10只裸鼠:空白组肿瘤接种部位旁皮下注射0.2 ml 0.9%Na Cl溶液、空载体组注射0.2 ml含100μg PAMAM-Lac Z质粒的溶液、给药组注射0.2 ml含100μg PAMAM-NK4质粒的溶液、阳性对照组腹腔注射0.2 ml多柔比星(100μg)溶液。连续注射7 d,第30天处死裸鼠,完整摘除肿瘤,测量肿瘤体积和质量。Western blotting检测各组移植瘤组织NK4蛋白表达。结果:PAMAM-NK4纳米粒转染MDA-MB-231、MCF-7细胞后能够稳定表达NK4蛋白、抑制细胞增殖和增加细胞凋亡率。成功建立MDA-MB-231细胞移植瘤裸鼠模型,给药组和阳性对照组肿瘤体积及质量显著低于空白组(P<0.05),且安全性良好;给药组NK4蛋白表达显著高于空白组(P<0.05)。结论:PAMAMNK4纳米粒对乳腺癌MDA-MB-231、MCF-7细胞生长具有抑制作用,并对人乳腺癌细胞移植瘤裸鼠模型具有治疗作用。
OBJECTIVE: To investigate the inhibitory effect of NK4 gene mediated by polyamidoamine dendrimer (PAMAM) on the growth of breast cancer cells MDA-MB-231 and MCF-7 and the effect on the growth of MDA-MB-231 Therapeutic effect of transplanted tumor in nude mice model. Methods: PAMAM-NK4 nanocomposite particles, nanocomplexes and empty vector PAMAM were transfected into MDA-MB-231 and MCF-7 cells respectively as experimental group and control group. MTT assay, Western blotting and flow cytometry were used to detect the effects of PAMAM-NK4 transfection on cell proliferation, NK4 protein expression and apoptosis respectively. Forty nude mice were transplanted subcutaneously into nude mice model of MDA-MB-231 cells and randomly divided into 4 groups with 10 mice in each group: 0.2 ml 0.9% Na Cl solution was injected subcutaneously into the tumor site of the blank group, The vehicle group was injected with 0.2 ml of a solution containing 100 μg of PAMAM-Lac Z plasmid. The treatment group was injected with 0.2 ml of a solution containing 100 μg of PAMAM-NK4 plasmid. The positive control group was intraperitoneally injected with 0.2 ml doxorubicin (100 μg) solution. After 7 days of continuous injection, the nude mice were sacrificed on the 30th day, the tumor was completely removed and the tumor volume and quality were measured. Western blotting was used to detect the expression of NK4 protein in each group. RESULTS: After transfected with PAMAM-NK4 nanoparticles, MDA-MB-231 and MCF-7 cells could stably express NK4 protein, inhibit cell proliferation and increase apoptosis rate. Successfully established MDA-MB-231 cell xenografts in nude mice model, the tumor volume and quality of the drug-treated group and the positive control group was significantly lower than the blank group (P <0.05), and the safety is good; NK4 protein expression was significantly higher Blank group (P <0.05). CONCLUSION: PAMAMNK4 nanoparticles can inhibit the growth of breast cancer MDA-MB-231 and MCF-7 cells and have a therapeutic effect on human breast cancer xenografts in nude mice.