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机构地区:[1]吉林大学中日联谊医院,吉林长春130013 [2]吉林医药学院检验学院,吉林吉林132013
出 处:《北华大学学报(自然科学版)》2016年第3期335-338,共4页Journal of Beihua University(Natural Science)
基 金:吉林省教育厅科学技术研究项目(2012494)
摘 要:目的探讨抑癌基因IL-24联合大蒜素对肺癌A549细胞凋亡的影响,并初步探讨其作用机制.方法提取前期构建的p DC316-h IL-24-EGFP质粒,转染肺癌A549细胞,分为A组(空白对照组)、B组(单独大蒜素组(40μL/m L))、C组(脂质体+IL-24组)、D组(大蒜素(40μL/m L)+脂质体+IL-24).荧光显微镜下观察质粒的转染情况;流式细胞仪检测细胞凋亡;Western blot法检测p DC316-h IL-24-EGFP、大蒜素和联合用药作用A5 4 9细胞4 8 h后IL-24,Bax,Bcl-2和Caspase-3蛋白表达水平.结果大蒜素(40μL/m L)和p DC316-h IL-24-EGFP单用48 h对A549细胞均有明显抑制作用;大蒜素(40μL/m L)联合p DC3 1 6-h IL-24-EGFP比单用p DC3 1 6-h IL-24-EGFP作用更强(P<0.05);大蒜素(40μL/m L)、p DC316-h IL-24-EGFP和两药联用48 h后,A549细胞凋亡率分别为32.7%,39.4%和68.8%;大蒜素(40μL/m L)和/或IL-24作用于A549细胞48 h后,Bax/Bcl-2的比值及Caspase-3表达均上调,联合用药组效果尤其明显.结论p DC316-h IL-24-EGFP联合大蒜素可协同抑制人肺癌A549细胞生长,机制可能是通过调节基因Bax/Bcl-2的表达比率,并上调Caspase-3的表达,进而诱导A549细胞的凋亡.Objective To investigate the apoptosis effect of tumor suppressor gene IL-24 combined with allicin on A549 lung cancer cell lines,as well as to explore the molecular mechanisms.Method Pre-constructed recombinant plasmid p DC316-h IL-24-EGFP was extracted and then transfected to A549 cells.There were four test groups treated with different agents,including group A(blank control),group B(allicin(40 μL/m L)),group C(p DC316-h IL-24-EGFP and lipofectamine transfection),group D(p DC316-h IL-24-EGFP and lipofectamine transfection combined with allicin(40 μL/m L)).The transduction efficiency of target cells was detected by fluorescent microscope,and apoptosis was tested by flow cetometry.The expression levels of IL-24,Bax,Bcl-2 and Caspase-3 in A549 cells treated with p DC316-h IL-24-EGFP and allicin alone or in combination for 48 h were detected by Western blot.Results Both allcin(40 μL/m L) and p DC316-h IL-24-EGFP alone could inhibit thegrowth of A549 lung cancer cells,and the combined treatment resulted a synergistic effect(P〈0.05).After being treated with allcin(40 μL/mL),p DC316-h IL-24-EGFP and combination therapy for 48 h,the apoptosis rates of A549 cells was 32.7%,39.4% and 68.8%,respectively.The rate of Bax/Bcl-2 and the expressions of Caspase-3in the treatment groups all increased,and the combination groups exhibited the most obvious effect.Conclusion The combination of p DC316-h IL-24-EGFP and allcin has a synergistic inhibition on the growth of A549 cells,which apoptosis mechanism may be related to the increased ratio of Bax/Bcl-2 and the regulation of Caspase-3 protein expression.
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