SPINK6蛋白在人乳腺癌细胞MCF-7中的表达与定位研究

Expression and Location of SPINK6 in Human Breast Cancer Cell MCF-7

为探讨Kazal型丝氨酸蛋白酶抑制剂6(SPINK6)蛋白的表达定位与其抗肿瘤作用之间的关系,本文通过免疫荧光的方法研究SPINK6蛋白在不同细胞系中的表达与定位,并构建产生pEGFPN1-SPINK6载体转染至乳腺癌细胞系MCF-7中,使用ER-Tracker标记内质网、lyso-Tracker标记溶酶体、Dil标记细胞膜,共聚焦显微镜观察SPINK6-GFP融合蛋白的定位.结果表明,天然状态的SPINK6蛋白主要分布在细胞质和细胞膜上,并在细胞质中有一定的聚集.对照载体pEGFP-N1转染的细胞表达的绿色荧光主要分布在细胞核中,细胞质中也有均匀分布,而SPINK6-GFP融合蛋白聚集于内质网和细胞膜上.说明SPINK6蛋白在合成后被分泌到细胞膜外发挥其作用,可能与抗炎症与抗肿瘤转移相关.

Abstract： To reveal the relationship between the location of Kazal type serine protease inhibitor 6（SPINK6） and its function of anti-tumor. The expression and location of SPINK6 protein were studied by immunofluorescenee. The pEGFPN1-SPINK6 vector was constructed and transfected into MCF-7 cell line, then green fluorescent protein （GFP） was observed together with other organelle markers, including ER-trader, lyso-trader, Dil, by the laser scanning confocal microscopy. Results showed that SPINK6 was mainly located in the cytoplasm and on the cell membrane in natural state. It was showed that green fluorescence protein was distributed in the cell nucleus transfected with pEGFP-N1 vector, but mainly distributed on the plasma membrane and ER transfected with pEGFPN1-SPINK6 vector. The results suggested that SPINK6 protein might be secreted to the outside space of the cell after production, and played roles of anti-inflammatory and anti-tumor functions.