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目的 :研究人乳腺癌细胞BT474中RNPC1对Snail表达的调控,及其与乳腺癌淋巴结转移等生物学行为的关系。方法:采用免疫组化(SP法)检测42例伴有淋巴结转移及39例无淋巴结转移的乳腺癌组织中RNPC1蛋白的表达,并计算差值;通过TCGA数据库分析伴有或不伴有淋巴结转移的乳腺癌组织中RNPC1基因表达差异;采用慢病毒转染法干扰或过表达RNPC1基因,实时荧光定量PCR(qRT-PCR)和Western blot法检测BT474细胞中Snail的表达量;以双荧光素酶报告基因实验证实RNPC1与靶基因Snail的结合。结果:与有淋巴结转移的乳腺癌组织相比,RNPC1的表达量在无淋巴结转移的组织中相对较高;干扰RNPC1能够明显增加BT474细胞中Snail的表达,过表达RNPC1可以显著降低Snail的表达;RNPC1通过结合Snail3′-非编码区(3′-UTR)降低其表达水平。结论 :RNPC1可以通过降低Snail的表达,影响乳腺癌的转移能力。
AIM: To investigate the regulation of Snail expression in human breast cancer cell line BT474 and its relationship with the biological behavior of lymph node metastasis in breast cancer. Methods: The expressions of RNPC1 protein in 42 cases of breast cancer with lymph node metastasis and 39 cases of non-lymph node metastasis were detected by immunohistochemistry (SP method), and the differences were calculated. The TCGA database was used to analyze the expression of RNPC1 protein with or without lymph node metastasis The expression of RNPC1 gene in BT474 cells was detected by qRT-PCR and Western blot. The expression of RNPC1 gene was detected by double-luciferase Reporter gene experiments confirmed the binding of RNPC1 to the target gene Snail. Results: Compared with breast cancer with lymph node metastasis, the expression of RNPC1 was higher in the tissues without lymph node metastasis. Interference with RNPC1 could significantly increase the expression of Snail in BT474 cells. The over-expression of RNPC1 could significantly reduce the expression of Snail. RNPC1 decreases its expression level by binding to the Snail3’-non-coding region (3’-UTR). Conclusion: RNPC1 can affect the metastasis of breast cancer by decreasing the expression of Snail.