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目的:检测乳腺癌细胞株MCF-7中瘦素受体(leptin receptor)的表达,并探讨瘦素(leptin,LEP)对他莫昔芬(tamox-ifen,TAM)抑制MCF-7细胞增殖的影响及机制。方法:用免疫荧光及Western blot法检测MCF-7细胞中瘦素受体的表达。以100 ng/ml的LEP及1 000 nmol/L的TAM同时处理MCF-7细胞,用MTT法检测LEP及TAM对细胞增殖的影响,并用West-ern blot及免疫荧光的方法检测作用前后雌激素受体(ERα)表达水平的变化。合成人雌激素反应元件(estrogen response ele-ment,ERE)的核心片段,并将其插入pGL3-promoter载体的多克隆位点,构建成含有ERE报告基因的质粒pERE-LUC,用脂质体转染的方法将该质粒瞬转到MCF-7细胞中,用LEP及TAM处理后检测荧光素酶的表达,以探讨其对ERα转录的影响。结果:MCF-7细胞呈瘦素受体的阳性表达。100 ng/ml的LEP能够削弱TAM对MCF-7细胞的增殖抑制作用,并能够通过上调ERα的转录及表达水平来干扰TAM对细胞增殖的抑制作用。结论:LEP能够干扰TAM抑制乳腺癌细胞增殖的作用,LEP与乳腺癌患者TAM治疗的疗效可能存在着一定的关系。
OBJECTIVE: To detect the expression of leptin receptor (MCF-7) in breast cancer cell line MCF-7 and investigate the effect of leptin (LEP) on the proliferation of MCF-7 cells induced by tamoxifen Impact and mechanism. Methods: The expression of leptin receptor in MCF-7 cells was detected by immunofluorescence and Western blot. MCF-7 cells were treated with 100 ng / ml LEP and 1000 nmol / L TAM at the same time. The effects of LEP and TAM on cell proliferation were detected by MTT assay. The levels of estrogen and estradiol were measured by West-ern blot and immunofluorescence Receptor (ERα) expression level changes. The core fragment of human estrogen response ele-ment (ERE) was synthesized and inserted into the multiple cloning site of the pGL3-promoter vector to construct the plasmid pERE-LUC containing the ERE reporter gene. The plasmid was transiently transfected into MCF-7 cells, and the expression of luciferase was detected by LEP and TAM treatment to explore its effect on ERα transcription. Results: MCF-7 cells showed positive expression of leptin receptor. LEP at 100 ng / ml could attenuate the inhibitory effect of TAM on the proliferation of MCF-7 cells and interfere with the inhibitory effect of TAM on cell proliferation by up-regulating the transcription and expression of ERα. CONCLUSION: LEP can interfere with the inhibitory effect of TAM on the proliferation of breast cancer cells. LEP may play a role in the therapeutic effect of TAM in breast cancer patients.