论文部分内容阅读
目的:探讨黑素瘤相关抗原-A9(melanoma-associated antigen,MAGE-A9)对人乳腺癌细胞中P53转录活性及功能的影响。方法:通过LipofectamineTM2000体外转染质粒pcDNA3.0、pcDNA3.0-p53、pCMV6-MAGE-A9和pcDNA3.0-p53/MAGE-A9至人乳腺癌MDA-MB-231细胞,RT-PCR和Western blotting检测细胞中p21WAF1mRNA和蛋白的表达,荧光素酶报告基因分析检测细胞中p21WAF1启动子介导的荧光素酶表达活性,MTT法检测转染不同质粒对MDA-MB-231细胞增殖的影响。结果:转染pcDNA3.0-p53/MAGE-A9组MDA-MB-231细胞中p21WAF1mRNA和蛋白表达水平均明显低于pcDNA3.0-p53组[(0.15±0.01)vs(0.18±0.02),(0.03±0.00)vs(0.06±0.01);均P<0.05]。转染pcDNA3.0-p53质粒可以增强MDAMB-231细胞中p21WAF1启动子介导的荧光素酶的表达[(58.56±3.47)vs(1.00±0.12),P<0.01],转染pcDNA3.0-p53/MAGE-A9后,MDA-MB-231细胞中p21WAF1启动子介导的荧光素酶的表达较转染pcDNA3.0-p53组明显降低[(22.02±4.91)vs(58.56±3.47),P<0.05]。与pcDNA3.0组相比,pcDNA3.0-p53组MDA-MB-231细胞增殖率明显明显降低[(228.89±22.39)%vs(337.23±23.67)%,P<0.05];而pcDNA3.0-p53/MAGE-A9组MDA-MB-231细胞增殖率明显高于pcDNA3.0-p53组[(291.51±5.91)%vs(228.89±22.39)%,P<0.05]。结论:MAGE-A9可抑制MDA-MB-231细胞中P53的转录活性及细胞增殖。
Objective: To investigate the effect of melanoma-associated antigen (MAGE-A9) on the transcriptional activity and function of P53 in human breast cancer cells. METHODS: Human breast cancer MDA-MB-231 cells were transfected with pcDNA3.0, pcDNA3.0-p53, pCMV6-MAGE-A9 and pcDNA3.0-p53 / MAGE-A9 by LipofectamineTM2000 in vitro. RT- The expression of p21WAF1 mRNA and protein was detected by luciferase reporter assay. The luciferase activity was detected by luciferase reporter assay. The effect of p21WAF1 promoter on the proliferation of MDA-MB-231 cells was detected by MTT assay. Results: The expression of p21WAF1 mRNA and protein in pcDNA3.0-p53 / MAGE-A9 group was significantly lower than that in pcDNA3.0-p53 group (0.15 ± 0.01 vs 0.18 ± 0.02, 0.03 ± 0.00) vs (0.06 ± 0.01), all P <0.05]. The pcDNA3.0-p53 plasmid transfected pcDNA3.0-p53 plasmid could enhance the expression of p21WAF1 promoter in MDAMB-231 cells (58.56 ± 3.47 vs 1.00 ± 0.12, P <0.01) p53 / MAGE-A9, the expression of p21WAF1 promoter in MDA-MB-231 cells was significantly lower than that of pcDNA3.0-p53 group [(22.02 ± 4.91 vs 58.56 ± 3.47, P <0.05]. The proliferation rate of MDA-MB-231 cells in pcDNA3.0-p53 group was significantly lower than that in pcDNA3.0 group [(228.89 ± 22.39)% vs (337.23 ± 23.67)%, P <0.05] The proliferation rate of MDA-MB-231 cells in p53 / MAGE-A9 group was significantly higher than that in pcDNA3.0-p53 group [(291.51 ± 5.91)% vs (228.89 ± 22.39)%, P <0.05]. Conclusion: MAGE-A9 can inhibit the transcriptional activity and proliferation of P53 in MDA-MB-231 cells.