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.
出处
《北华大学学报（自然科学版）》
CAS
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)
关键词
IL-24基因
大蒜素
联合用药
A549细胞
细胞凋亡
IL-24 gene
allcin
combination drug therapy
human lung adenocarcinoma A549 cell
apoptosis
