趋化因子受体CXCR6在乳腺癌细胞系中的表达及其意义

Expression of Chemokine Receptor CXCR6 and Its Significance in Breast Cancer Cell Lines

目的探讨趋化因子受体(CXCR6)在不同侵袭性乳腺癌细胞系和正常乳腺上皮细胞株的表达及干扰表达后与癌细胞恶性表型的关系。方法采用Real time-PCR和Western blot分别检测不同侵袭性乳腺癌细胞系(SK-BR-3、MCF-7、MDA-MB-231)和正常乳腺上皮细胞(MCF-10A)中CXCR6mRNA和蛋白质的表达;采用慢病毒RNA干扰技术沉默MDA-MB-231乳腺癌细胞系中CXCR6的表达,并利用噻唑蓝(MTT)实验、Transwell小室、Real time-PCR〔检测血管内皮生长因子(VEGF)表达〕研究CXCR6在MDA-MB-231乳腺癌细胞中的作用。结果正常乳腺上皮细胞MCF-10A中CXCR6表达最低,不同侵袭性的乳腺癌细胞系中CXCR6的表达不同,高侵袭性乳腺癌细胞MDA-MB-231明显较低侵袭性乳腺癌细胞SK-BR-3和MCF-7表达高(P<0.05)。通过慢病毒RNA干扰技术沉默CXCR6在MDA-MB-231细胞中的表达后,细胞的体外增殖能力、侵袭能力和VEGF表达水平降低(P均<0.05)。结论 CXCR6在乳腺癌细胞株中的表达水平与细胞的侵袭性有关,干预癌细胞中的CXCR6表达将可能降低癌细胞的体外恶性生物学行为。

Objective To detect the expression of Chemokine receptor CXCR6 in invasive breast cancer cell lines and normal mammary epithelial cell line, and assess the relationship between CXCR6 expression and malignant behavior of breast cancer cells. Methods Expression level of CXCR6 in different invasive breast cancer cell lines （SK-BR-3,MCF-7,MDA-MB-231） and normal mammary epithelial cell line （MCF-10A）was detected by real time reverse transcription-polymerase chain reaction （real time-PCR） and Western blot. Lentivirus was employed to interfere CXCR6 expression in MDA-MB-231. MTT assay and transwell chamber were used to study proliferative and invasive ability of those cells respectively. Vascular enothelial growth factor （VEGF） expression was detected to study the role of CXCR6 in angiogenesis. Results At both mRNA level and protein level, normal mammary epithelial cell line MCF-10A showed the weakest CXCR6 expression. The breast cancer cell lines expressed CXCR6 in different levels, the expression level of CXCR6 in highly invasive cell line MDA-MB-231 was significantly higher than that in two low-invasive cell lines SK-BR-3 and MCF-7 （P〈0. 05）. Silencing CXCR6 gene by Lentivirusmediated RNA interference in MDA-MB-231 inhibited its proliferation ability, invasion ability and angiogenesis ability in vitro （P〈0. 05）. Conclusion Different invasive breast cancer cell lines express CXCR6 at different levels, positively correlated with its invasive ability.