摘要
目的探讨IL-24基因转染乳腺癌MDA-MB-231细胞的增殖抑制和促凋亡作用。方法利用脂质体将穿梭质粒pDC316-hIL-24-EGFP瞬时转染至乳腺癌MDA-MB-231细胞,通过RT-PCR检测转染后hIL-24基因mRNA的转录,Western blot检测转染后IL-24和caspase-3蛋白的表达,MTT比色法测定细胞增殖的抑制,流式细胞仪检测细胞的凋亡和细胞周期的变化,Hoechst 33258染色检测细胞凋亡。结果 IL-24基因可在MDA-MB-231细胞中成功转录及表达。IL-24可上调MDA-MB-231细胞中caspase-3蛋白表达。IL-24基因的表达使得乳腺癌MDA-MB-231细胞出现增殖抑制。流式细胞仪检测显示MDA-MB-231细胞DNA合成受到抑制,细胞周期主要抑制在G2/M期。Hoechst 33258染色显示IL-24基因转染后MDA-MB-231细胞出现凋亡。结论 IL-24基因转染乳腺癌MDA-MB-231细胞后可能通过上调caspase-3的表达而抑制细胞增殖,促进其凋亡。
Objective To explore the effect of IL-24 gene on proliferation and apoptosis of human breast cancer cell line MDA-MB-231. Methods The recombinant shuttle vector pDC316-hlL-24-EGFP was transiently transfeeted in- to human breast cancer cell line MDA-MB-231 with lipofectamine. Both IL-24 mRNA and protein were detected by RT-PCR and Western blot respectively. The inhibition of cell proliferation was evaluated by MTF assay. Changes in cell cycle were detected by flow cytometry （ FCM ）. The cell apoptosis was studied by Hoechst 33258 staining. Results IL-24 was successfully transcribed and expressed in MDA-MB-231 cells. IL-24 enhanced protein caspase- 3 expression level of MDA-MB-231 cells. The IL-24 expression inhibited MDA-MB-231 cell proliferation. In MDA- MB-231 cells of IL-24 over-expressed the DNA synthesis was inhibited and arrested in G2/M phrase according to FCM. Increased apoptosis of MDA-MB-231 cells was observed by Hoeehst33258 staining. Conclusions IL-24 gene overexpressed is able to effectively inhibit the proliferation and promote the apoptosis of breast cancer cells, which may be explained by enhancing of protein caspase-3 level.
出处
《基础医学与临床》
CSCD
北大核心
2012年第7期793-797,共5页
Basic and Clinical Medicine
基金
吉林医药学院青年科研基金(2009-04)
作者简介
通信作者（correspondingauthor）：wenyuezhuang@163．com
