【摘要】 目的研究CC族趋化因子结合蛋白D6在乳腺癌中的组成性表达及细胞因子IL-1β诱导后D6表达的改变。方法采用半定量RT-PCR法检测D6mRNA在人乳腺癌细胞株、乳腺癌组织以及正常脾组织中的表达,并用实时荧光定量-PCR进一步验证其在细胞中表达的差异;Westernblot法分析D6蛋白在乳腺癌细胞中的表达;用重组人IL-1β(0.5ng/mL,1ng/mL,2ng/mL)刺激MDA-MB-231细胞24h,实时荧光定量-PCR分析D6基因表达的改变。结果D6mRNA在所有9种人乳腺癌细胞系、4例乳腺癌组织以及作为阳性对照的人正常脾组织中均可检出,其表达水平因细胞的不同而异,其中MCF-7、ZR-75-1和SK-BR-3细胞的表达量较高。MDA-MB-435和MDA-MB-231细胞中可检测出D6蛋白。此外,以MDA-MB-231细胞为对象的细胞因子诱导实验结果显示,IL-1β可呈剂量依赖性地促进D6的基因表达。结论D6分子在来源于不同患者的乳腺癌组织及生物学特性各异的多种人乳腺癌细胞系中均呈组成性表达,提示CC族趋化因子的结合蛋D6可能参与乳腺癌中趋化因子的调控,从而影响乳腺癌的发生、发展过程。更多还原

【Abstract】 Objective: To investigate the constitutive gene expression of chemokine binding protein D6 in breast cancer and its regulation by interleukin-1 (IL-1)β. Methods: D6 mRNAs in 9 cases of human breast cancer cell lines, 4 cases of breast cancer tissues, and normal human spleen tissues were analyzed by semi-quantitative RT-PCR. The difference of D6 gene expression was further confirmed by real-time PCR. Western blot was used to detect D6 protein expression in breast cancer cells. In addition, the dynamics of D6 mRNA in MDA-MB-231 cells was monitored after stimulation with recombinant IL-1β at 0.5, 1, and 2 ng/mL for 24 h. Results: D6 mRNAs can be detected in all of 9 breast cancer cell lines, 4 breast cancer tissue and normal human spleen which served as a positive control with different relative levels. D6 mRNA expression was specifically prominent in MCF-7, ZR-75-1, and SK-BR-3 cells. D6 protein could also be detected in MDA-MB-435 and MDA-MB-31 breast cancer cells. The cytokine irritation experiment in MDA-MB-231 cells revealed that IL-1β enhanced D6 mRNA expression in a dose-dependent manner. Conclusion: This study demonstrates that D6 mRNA is constitutively expressed in breast cancer tissues from different patients and multiple human breast cancer cell lines with different biological characteristics, suggesting that this chemokine binding protein may also play a role in chemokine network which influence the development and progression of breast cancer.更多还原