近红外七甲川花菁染料IR-80染色鉴别乳腺良恶性细胞

Identification of breast cancer cells from benign breast cells with a NIR fluorescence dye IR-80

目的观察乳腺上皮细胞株和乳腺癌细胞株通过近红外(near-infrared,NIR)荧光染料IR-80活体染色后不同的染色效果,探讨最佳实验条件用于良、恶性乳腺细胞的鉴别。方法将人乳腺癌细胞株MCF-7、MDA-MB231和人乳腺上皮细胞株MCF-10A与5、2.5、1.25、0.625μmol/L浓度的NIR荧光染料IR-80在37℃下共同孵育30 min,缓冲液清洗后,应用激光共聚焦显微镜观察乳腺癌细胞和乳腺上皮细胞在500 ms、1 s和2 s曝光条件下荧光强度。结果当曝光时间一定时,随着IR-80染料应用液浓度的降低,每种细胞系NIR荧光强度逐渐降低;当IR-80染料应用液浓度一定时,随着曝光时间的增加,NIR荧光强度有所增强,但不明显;同样浓度和曝光时间下观察,乳腺癌细胞系(MCF-7、MDA-MB231)荧光染色强度高于人乳腺上皮细胞系(MCF-10A)。当IR-80应用液浓度为1.25μmol/L时,不同曝光条件下,乳腺癌细胞系NIR荧光染色强度明显高于乳腺上皮细胞系,而乳腺上皮细胞系不发荧光。结论七甲川花菁染料IR-80活体染色时,乳腺癌细胞荧光强度明显高于乳腺上皮细胞,当IR-80染料应用浓度为1.25μmol/L,曝光时间为1 s可鉴别乳腺良、恶性细胞,其有望成为鉴别临床乳腺穿刺标本和手术新鲜组织标本良、恶性的新方法。

Purpose To investigate the staining characteristics of new near-infrared （NIR） fluorescence dyes IR-80 in breast epithelial cell line and breast cancer cell line, so as to explore the best experimental conditions for the identification of benign and malignant breast cells. Methods Human breast cancer lines （MDA-MB231 and MCF-7 ） and human breast epithelial cell line （MCF-10A） were evaluated. NIR staining intensity of IR-80 in 500 ms, 1 s and 2 s exposure time was observed with incubation concentration of 5, 2. 5, 1.25, 0. 625 μmol/L. The incubation condition was 37℃ and 30 rain. Results The fluorescence intensity of each kind of cells decreased gradually with the decreased concentration of IR-80 dye solution when the exposure time was certain. Meanwhile, with in- creasing of the NIR exposure time, the fluorescence intensity increased, but not obvious when the concentration of IR-80 dye solution was certain. Under the same concentration of IR-80 and exposure time, the fluorescence intensity of breast cancer cells （MCF-7, MDA-MB231 ） was significantly higher than that of human breast epithelial cells （ MCF-IOA）. When the IR-80 concentration was 1.25 μmol/L and in different exposure time, the fluorescence intensity of breast cancer cell line was significantly higher than that of breast epithelial cell line while the breast epithelial cell line did not emit fluorescence. Conclusion IR-80 live staining intensity in breast cancer cells is significantly higher than that in breast epithelial cells. It can differentiate breast benign and malignant cells when the IR- 80 concentration is 1.25 μmol/L and the exposure time is 1 s. This study may provide a new method for the identification of benign and malignant breast biopsy specimens and fresh tissue samples.