IL-18基因修饰人脐带间充质干细胞对乳腺癌细胞系增殖的影响

Effect of interleukin-18 gene modified human umbilical cord mesenchymal stem cells on proliferation of breast cancer cell

目的：建立携带白细胞介素18（IL-18）基因人脐带间充质干细胞（HUMSCs）株并探讨其对乳腺癌细胞增殖的影响。方法分离培养HUMSCs，慢病毒介导IL-18基因转染HUMSCs建立携带 IL-18基因人脐带间充质干细胞（ IL-18-HUMSCs ）株，半定量 RT-PCR 及 Western 印迹检测IL-18mRNA及蛋白表达情况。 Transwell体系共培养后第1、3、5天用Cell Counting Kit-8（CCK-8）试剂盒检测MCF-7细胞增殖活力，绘制细胞生长曲线。结果 IL-18-HUMSCs能稳定高效表达IL-18基因，并有效抑制乳腺癌细胞增殖。其 IL-18 mRNA 相对表达量为：实验组（ IL-18-HUMSCs ）1.40±0.21，阴性对照组（NV-HUMSCs）0.59±0.09，空白对照组（HUMSCs）0.71±0.05；与各对照组比较，差异有统计学意义（F＝31.81，P＝0.001）。 IL-18蛋白相对表达量为：实验组（IL-18-HUMSCs）1.54±0.27，阴性对照组（NV-HUMSCs）0.57±0.04，空白对照组（HUMSCs）0.59±0.23；与各对照组比较，差异有统计学意义（ F＝22.32， P＝0.002）。共培养后第5天，乳腺癌细胞增殖受到明显抑制。结论HUMSCs是IL-18基因高效稳定表达载体，IL-18-HUMSCs可有效抑制乳腺癌细胞增殖。

Objective To establish human umbilical cord mesenchymal stem cells （ HUMSCs ） strain transfected with interleukin-18 （ IL-18 ） gene and examine its effects on the proliferation of breast cancer cell （ MCF-7 ）.Methods HUMSCs were isolated and cultured.And the lentivirus-IL-18 vector containing human IL-18 gene was constructed and transfected into HUMSCs.The expressions of IL-18 gene mRNA and protein were detected by semi-quantitative reverse transcription-polymerase chain reaction （ RT-PCR） and Western blot.After co-culturing for 1,3,5 days,Transwell and cell counting kit-8（CCK-8）assays were performed on MCF-7 to plot the cell growth curve.Results IL-18-HUMSCs could stably express of IL-18 gene and inhibit the proliferation of breast cancer cells.The IL-18 mRNA relative expression amounts were 1.40 ±0.21 for experimental group （ IL-18-HUMSCs ） , 0.59 ±0.09 for negative control group （ NV-HUMSCs） and 0.71 ±0.05 for blank control group （ HUMSCs ）.As compared with control group , the difference was statistically significant （ F=31.81,P=0.001）.The relative expressions of IL-18 protein were 1.54 ±0.27 for experimental group （ IL-18-HUMSCs ） , 0.57 ±0.04 for negative control group （ NV-HUMSCs） and 0.59 ±0.23 for blank control group （ HUMSCs ）.As compared with control group , the difference was statistically significant （ F =22.32, P =0.002 ）.After co-culturing for 5 days, the cellular proliferation was significantly inhibited.Conclusion IL-18 gene has been successfully transfected into HUMSCs and has a stable expression.And IL-18-HUMSCs can effectively inhibit the proliferation of breast cancer cells in vitro.