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目的:探讨蛋白质精氨酸甲基转移酶2(protein arginine methyltransferase2,PRMT2)基因在多种乳腺癌细胞株中的表达情况,以及外源性PRMT2基因过表达对乳腺癌SKBR-3细胞生长特性的影响。方法:采用real-time RT-PCR法检测PRMT2基因在不同乳腺癌细胞株中的表达,并建立稳定表达pcDNA3.1/NT-GFP-PRMT2的SKBR-3细胞株;激光共聚焦显微镜下观察外源性PRMT2蛋白在细胞中的定位,检测在雌激素和雌激素受体(estrogen receptor,ER)拮抗剂4-OHT作用下过表达PRMT2基因对SKBR-3细胞增殖的影响。结果:PRMT2基因在ERα阳性乳腺癌细胞株中表达水平明显高于ERα阴性乳腺癌细胞株;在转染PRMT2基因的SKBR-3细胞株中,雌激素反应元件-荧光素酶报告基因(estrogen response elements-luciferasereporter,ERE-luc)的转录活性明显升高。在无处理因子情况下,外源基因PRMT2在SKBR-3细胞中的表达对细胞的形态及生长速度无明显影响。与未转染及转染GFP空载体的SKBR-3细胞相比,稳定转染PRMT2基因的SKBR-3细胞对雌激素的敏感性明显下降,但其对4-OHT处理的敏感性降低无明显差异。结论:PRMT2基因表达的多少及部位与乳腺癌细胞中ERα的表达密切相关。外源性PRMT2基因在乳腺癌SKBR-3细胞中稳定表达使细胞对雌激素的敏感性降低,但不增加细胞对ER拮抗剂4-OHT的耐药性。
Objective: To investigate the expression of protein arginine methyltransferase 2 (PRMT2) gene in many breast cancer cell lines and the effect of exogenous PRMT2 gene overexpression on the growth of breast cancer SKBR-3 cells influences. Methods: The expression of PRMT2 gene in different breast cancer cell lines was detected by real-time RT-PCR and the SKBR-3 cell line stably expressing pcDNA3.1 / NT-GFP-PRMT2 was established. Under confocal laser scanning microscope To investigate the effect of overexpression of PRMT2 gene on the proliferation of SKBR-3 cells induced by estrogen and estrogen receptor (ER) antagonist 4-OHT. Results: The expression of PRMT2 gene in ERα positive breast cancer cell lines was significantly higher than that in ERα negative breast cancer cell lines. In the SKBR-3 cell line transfected with PRMT2 gene, estrogen response (estrogen response) elements-luciferasereporter, ERE-luc) significantly increased the transcriptional activity. In the absence of treatment factors, the expression of exogenous gene PRMT2 in SKBR-3 cells had no significant effect on cell morphology and growth rate. SKBR-3 cells stably transfected with PRMT2 gene showed a significant decrease in the sensitivity to estrogen compared with SKBR-3 cells transfected with GFP empty vector, but their sensitivity to 4-OHT treatment was not significantly reduced difference. Conclusion: The number and location of PRMT2 gene expression are closely related to the expression of ERα in breast cancer cells. Stable expression of exogenous PRMT2 gene in breast cancer SKBR-3 cells decreased the sensitivity of cells to estrogen, but did not increase the cell resistance to ER antagonist 4-OHT.