Sun Suyuan, Liu Xiaoyi, Jiang Dandan, Lü Zhidong, Li Funian
Diagnosis and Treatment Centre for Breast Disease, Affiliated Hospital of Medical College, Qingdao University, Qingdao 266071, China.
Diagnosis and Treatment Centre for Breast Disease, Affiliated Hospital of Medical College, Qingdao University, Qingdao 266071, China. Email:
Zhonghua Yi Xue Za Zhi. 2014 Jul 8;94(26):2013-7.
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).
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.
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.
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.
构建白细胞介素-18(IL-18)基因转染的人脐带间充质干细胞(HUMSCs)株,并检测其对乳腺癌细胞(MCF-7)增殖的影响。
分离培养HUMSCs。构建含人IL-18基因的慢病毒-IL-18载体并转染至HUMSCs。采用半定量逆转录-聚合酶链反应(RT-PCR)和蛋白质印迹法检测IL-18基因mRNA和蛋白的表达。共培养1、3、5天后,对MCF-7进行Transwell实验和细胞计数试剂盒-8(CCK-8)检测,绘制细胞生长曲线。
IL-18-HUMSCs可稳定表达IL-18基因并抑制乳腺癌细胞增殖。实验组(IL-18-HUMSCs)IL-18 mRNA相对表达量为1.40±0.21,阴性对照组(NV-HUMSCs)为0.59±0.09,空白对照组(HUMSCs)为0.71±0.05。与对照组比较,差异有统计学意义(F=31.81,P=0.001)。实验组(IL-18-HUMSCs)IL-18蛋白相对表达量为1.54±0.27,阴性对照组(NV-HUMSCs)为0.57±0.04,空白对照组(HUMSCs)为0.59±0.23。与对照组比较,差异有统计学意义(F=22.32,P=0.002)。共培养5天后,细胞增殖明显受到抑制。
IL-18基因已成功转染至HUMSCs并稳定表达。IL-18-HUMSCs可在体外有效抑制乳腺癌细胞增殖。