Effect of RNA interference-mediated down-regulation of JMJD2A on human breast cancer cell line MCF-7

Abstract

Objective To study the effect of specific small interfering RNA (siRNA) silencing of Jumonji Domain Containing 2A (JMJD2A) on human breast cancer cell line MCF-7. Methods The chemically synthetic siRNA duplexes targeting JMJD2A was transfected into MCF-7 cells by HiPerFect Transfection Reagent. After siRNA interfering cells were harvested, the expression of JMJD2A was detected by real-time reverse transcription-polymerase chain reaction ( RT-PCR), and Western blotting. The activity of cell proliferation and cell cycle were examined by WST-8 and flow cytometry (FCM) respectively , and the abilities of cell invasion and metastasis were evaluated using Transv ell chambers. Results JMJD2A siRNA effectively inhibited the expression of JMJD2A mRNA and protein (P<0. 05). After siRNA interference, the cell proliferation was suppressed [ 1.45±0. 05 vs 1.73±0. 02(48h),P <0. 05] , and cell cycle was arrested at G0/G1 phase (53. 80±1. 80 vs 44. 84±1. 86,P<0. 05) with a significant decrease of cells in S phase (36. 55±1. 52 vs 47. 06±1. 26,P<0. 05). The ability of cell invasion and metastasis was depressed (P<0. 05). Conclusion siRNA interfering to JMJD2A effectively reverses some of the malignant biological characters of human breast cancer cell line MCF-7, implying that JMJD2A might be involved in the progression and metastasis of breast cancer cells. Key words: RNA interference; Breast carcinoma; Proliferation; Cell cycle; Metastasis