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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 investigate the effects of IL-24 gene on the proliferation and apoptosis of breast cancer MDA-MB-231 cells. Methods The shuttle plasmid pDC316-hIL-24-EGFP was transiently transfected into breast cancer cell line MDA-MB-231 by lipofectamine. The transcription of hIL-24 mRNA was detected by RT-PCR. The expression of IL-24 and caspase-3 protein were detected by MTT assay. The cell proliferation was measured by MTT colorimetric assay. The apoptosis and cell cycle were detected by flow cytometry. Hoechst 33258 staining was used to detect the apoptosis. Results IL-24 gene was successfully transcribed and expressed in MDA-MB-231 cells. IL-24 can up-regulate caspase-3 protein expression in MDA-MB-231 cells. The expression of IL-24 gene inhibits the proliferation of breast cancer MDA-MB-231 cells. Flow cytometry showed that the DNA synthesis of MDA-MB-231 cells was inhibited and the cell cycle was mainly inhibited in G2 / M phase. Hoechst 33258 staining showed apoptosis of MDA-MB-231 cells after transfection of IL-24 gene. Conclusion IL-24 gene may inhibit the proliferation and promote the apoptosis of breast cancer MDA-MB-231 cells by up-regulating the expression of caspase-3.