Department of Forensic Medicine, Shanghai Medical College, Fudan University, Shanghai 200032, PR China.
J Exp Clin Cancer Res. 2011 Oct 3;30(1):90. doi: 10.1186/1756-9966-30-90.
Previous data demonstrate that JMJD2A is a cancer-associated gene and may be involved in human breast cancer by demethylation of H3K9me3. The aim of this study was to investigate depressive effects on JMJD2A by transfection with JMJD2A-sepcific siRNA in human breast cancer cell line MDA-MB-231 and effects on cell proliferation, invasion and migration. JMJD2A-specific siRNA was chemically synthesised and transfected into human breast cancer cell line MDA-MB-231. Expression levels of JMJD2A were detected by quantitative real-time PCR and Western blot analysis. Cells proliferation was evaluated by using flow cytometric anlysis and MTT assay. The abilities of invasion and migration were evaluated by cell migration and invasion assay with Boyden chambers. The results showed that the transfection was successful and expression levels of JMJD2A mRNA and protein in siRNA group were both down-regulated. By MTT assay, the mean actual absorbance in siRNA group was significantly lower than that in blank control group (P < 0.05) and negative control group (P < 0.05). In addition, the percentage of cells in G0/G1 phase in siRNA group was significantly more than that in blank control group (P < 0.05) and negative control group (P < 0.05). Furthermore, by cell invasion and migration assay, the decreased number of migrated cells in siRNA group was observed (P < 0.05). These data imply that silencing JMJD2A gene could result in cell cycle change and proliferation inhibition, and lead to suppress tumor cell invasion and migration. It provides a new perspective in understanding the pleiotropic functions of JMJD2A and its contribution to human breast cancer.
先前的数据表明 JMJD2A 是一种与癌症相关的基因，可能通过 H3K9me3 的去甲基化参与人类乳腺癌的发生。本研究旨在通过转染 JMJD2A 特异性 siRNA 探讨其对人乳腺癌 MDA-MB-231 细胞系的抑郁作用及其对细胞增殖、侵袭和迁移的影响。化学合成 JMJD2A 特异性 siRNA 并转染入人乳腺癌 MDA-MB-231 细胞系。采用实时定量 PCR 和 Western blot 分析检测 JMJD2A 的表达水平。采用流式细胞术和 MTT 法评估细胞增殖。通过 Boyden 室细胞迁移和侵袭试验评估细胞侵袭和迁移能力。结果表明，转染成功，siRNA 组中 JMJD2A mRNA 和蛋白的表达水平均下调。MTT 试验结果显示，siRNA 组的平均实际吸光度明显低于空白对照组（P<0.05）和阴性对照组（P<0.05）。此外，siRNA 组中 G0/G1 期细胞的百分比明显高于空白对照组（P<0.05）和阴性对照组（P<0.05）。进一步通过细胞侵袭和迁移试验，观察到 siRNA 组中迁移细胞数量减少（P<0.05）。这些数据表明，沉默 JMJD2A 基因可导致细胞周期改变和增殖抑制，并导致肿瘤细胞侵袭和迁移受到抑制。这为理解 JMJD2A 的多效性功能及其对人类乳腺癌的贡献提供了新的视角。
