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目的探讨他莫昔芬(Tamoxifen,TAM)对子宫内膜癌RL-95-2细胞增殖及let-7g表达的影响。方法采用免疫荧光染色法检测RL-95-2细胞中雌激素受体(Estrogen receptor,ER)的表达;用不同浓度的TAM(1×10-11,1×10-9、1×10-7、1×10-5、1×10-4mol/L)处理RL-95-2细胞48和72 h,CCK-8法检测细胞的增殖活力;用1×10-7mol/L的TAM处理RL-95-2细胞72 h,流式细胞术检测细胞周期的变化;用不同浓度的TAM处理RL-95-2细胞72 h,实时荧光定量PCR(Real-time quantitative polymerase chain reaction,RT-qPCR)检测细胞中let-7g的表达,借助生物信息学软件预测let-7g的靶标基因,并对其生物学功能进行分析。结果 RL-95-2细胞中ER呈阳性表达;1×10-7mol/L的TAM处理RL-95-2细胞48 h及1×10-9～1×10-5mol/L的TAM处理细胞72 h,具有促进细胞增殖的作用,并使let-7g的表达量增加,1×10-7mol/L的TAM作用最为显著;TAM促使细胞从G0/G1期进入S期;let-7g作用的可能靶标基因为UHRF2、RB1、GAS7、PLAGL2、NAB1和ZNF282。结论适宜浓度的TAM对人子宫内膜癌RL-95-2细胞具有促增殖作用,并使细胞从G0/G1期进入S期,该作用可能部分是通过上调let-7g的表达而实现的。
Objective To investigate the effect of Tamoxifen on the proliferation and let-7g expression of endometrial carcinoma RL-95-2 cells. Methods The expression of estrogen receptor (ER) in RL-95-2 cells was detected by immunofluorescence staining. Tumor necrosis factor (ER) was detected with different concentrations of TAM (1 × 10-11, 1 × 10-9, 1 × 10-7 , 1 × 10-5 and 1 × 10-4mol / L) for 48 and 72 h respectively. The proliferation of RL-95-2 cells was detected by CCK-8 assay. RL-95-2 cells were treated with 1 × 10-7mol / 95-2 cells for 72 h, the changes of cell cycle were detected by flow cytometry; RL-95-2 cells were treated with different concentrations of TAM for 72 h, Real-time quantitative polymerase chain reaction (RT-qPCR) The expression of let-7g in cells was predicted by bioinformatics software and the biological function of let-7g was analyzed. Results RL-95-2 cells were positive for ER. TAM cells treated with 1 × 10-7 mol / L TAM for 48 h and 1 × 10-9 ~ 1 × 10-5 mol / L TAM-treated cells 72 h, with the promotion of cell proliferation, and let-7g expression increased, 1 × 10-7mol / L of TAM the most significant role; TAM to promote cells from the G0 / G1 phase S; let-7g role may The target genes are UHRF2, RB1, GAS7, PLAGL2, NAB1 and ZNF282. Conclusion TAM at a suitable concentration can promote the proliferation of human endometrial carcinoma RL-95-2 cells and lead the cells from the G0 / G1 phase to S phase. This effect may be partly caused by the up-regulation of let-7g expression.