天津医药

天津医药 ›› 2017, Vol. 45 ›› Issue (12): 1237-1241.doi: 10.11958/20170819

• 细胞与分子生物学 • 上一篇    下一篇

FOS 样抗原 1 对乳腺癌细胞 MDA-MB-231 增殖、 侵袭和 PRDM10 基因甲基化的影响

张晓宇 1， 康晓宁 2， 刘伟 1， 刘志明 1， 王鹏 1， 王遵义 1△   

  1. 作者单位： 1 河北省沧州市中心医院肿瘤外三科 （邮编 061001）， 2 超声二科 作者简介： 张晓宇 （1983）， 男， 硕士， 主治医师， 主要从事乳腺癌综合治疗的研究 △通讯作者 E-mail：czwzy99@163.com
  • 收稿日期:2017-07-21 修回日期:2017-10-24 出版日期:2017-12-15 发布日期:2017-12-15
  • 通讯作者: 张晓宇 E-mail:93956466@qq.com

Effects of FOSL1 on cell proliferation, cell invasiveness and the methylation of PRDM10 gene in breast cancer cell line MDA-MB-231

ZHANG Xiao-yu1, KANG Xiao-ning2, LIU Wei1, LIU Zhi-ming1, WANG Peng1, WANG Zun-yi1△   

  1. 1 Department of Oncological Surgery-3, 2 Department of Ultrasound-2, Cangzhou Central Hospital, Cangzhou 061001, China △Corresponding Author E-mail: czwzy99@163.com
  • Received:2017-07-21 Revised:2017-10-24 Published:2017-12-15 Online:2017-12-15

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摘要： 目的 构建 FOS 样抗原 1（FOSL1）基因沉默的载体， 研究 FOSL1 对人乳腺癌细胞系 MDA-MB-231 增殖 和侵袭的影响， 以及对此细胞中 PRDM10 基因的甲基化状态的影响。方法 FOSL1 沉默的 pLVX-shRNA-FOSL1- shRNA 干扰质粒载体， 采用慢病毒感染法分别用 pLVX-shRNA-FOSL1-shRNA 载体以及 pLVX-shRNA 空载的两种 病毒感染靶细胞 MDA-MB-231， 分别记作转染组和空载组。以未转染的 MDA-MB-231 作为对照组。RT-PCR 验证 FOSL1 表达后， 分别采用 MTT 实验和 Transwell 实验检测 MDA-MB-231 的细胞增殖能力和侵袭能力。采用 MSP 法 检测 PRDM10 基因的甲基化状态。Q-PCR 和 Western blot 分别检测 PRDM10 的 mRNA 和蛋白质表达水平。结果 MTT 实验显示， 24 h、 48 h 和 72 h 时， 转染组 MDA-MB-231 的 OD 值低于对照组 （均 P＜0.05）； 48 h 和 72 h 时， 转染 组 MDA-MB-231 的 OD 值低于空载组 （均 P < 0.05）。Transwell 实验显示， 与空载组和对照组相比， 转染组的 MDAMB-231 细胞侵袭能力降低 （均 P < 0.05）。MSP 实验显示， 与空载组和对照组相比， 转染组的 MDA-MB-231 细胞中 PRDM10 基因甲基化水平降低。Q-PCR 和 Western blot 实验显示， 与空载组和对照组相比， 转染组 PRDM10 的 mRNA 和蛋白质表达水平均升高。结论 FOSL1 基因沉默抑制 MDA-MB-231 细胞的增殖和侵袭， 可能与该细胞中 PRDM10 基因的去甲基化有关。

关键词: 乳腺肿瘤, 细胞增殖, 细胞侵袭, FOSL1, PRDM10

Abstract: Objective To construct the silencing vector of FOS like antigen 1 (FOSL1) gene, and study the effects of FOSL1 on cell proliferation, cell invasiveness and the methylation level of PRDM10 gene in breast cancer cell line MDAMB-231. Methods The FOSL1 silencing vector of gene pLVX-shRNA-FOSL1-shRNA was purchased. The FOSL1 silencing vector and the empty vector were separately transfected into MDA-MB-231, which were regarded as transfection group and empty group, respectively. Untransfected MDA-MB-231 was used as control group. FOSL1 was verified by PCR in MDA-MB-231. The cell proliferation ability and cell invasion ability of MDA-MB-231 were detected by MTT and Transwell assay, respectively. MSP was used to detect the methylation status of PRDM10 gene. The mRNA and protein expression levels of PRDM10 gene were detected by Q-PCR and Western-blot assay. Results MTT results showed that the optical density (OD) values were significantly lower in transfection group compared with those of control group at 24 h, 48 h and 72 h (all P<0.05), and the same as those compared with empty group at 48 h and 72 h (both P<0.05). Compared with empty group and control group, Transwell assays showed that the cell invasive abilities of MDA-MB-231 were decreased in transfection group (both P<0.05), and MSP assay showed that the methylation of PRDM10 gene was decreased in MDA-MB- 231, and Q-PCR and Western-blot tests showed that the expressions of PRDM10 gene were increased in mRNA level and in protein level. Conclusion Silencing of FOSL1 gene inhibits the proliferation and invasion of MDA-MB-231 cells, which might be related to the demethylation of PRDM10 gene in the cells.

Key words: breast neoplasms, cell proliferation, cell invasion, FOSL1, PRDM10