葡萄糖调节蛋白78在乳腺癌细胞侵袭及转移中的作用

The roles of GRP78 in the invasion and metastasis of breast cancer cells

目的探讨葡萄糖调节蛋白78(GRP78)在乳腺癌细胞(231)侵袭、转移过程所起的作用。方法构建GRP78真核反义表达载体,转染入231细胞,利用PCR和蛋白印迹方法检测转染有反义GRP78的231细胞和MCF-7细胞的GRP78表达情况;并用伤口愈合实验和体外侵袭实验证验转染细胞转移和侵袭的变化。结果转染反义GRP78的231细胞内GRP78RNA和蛋白表达明显低于未转染细胞;反义GRP78基因对乳腺癌细胞231细胞的侵袭和转移有抑制作用。结论GRP78是乳腺癌的一个促癌基因,下调GRP78基因表达能明显抑制乳腺癌细胞的侵袭和转移能力。GRP78为乳腺癌基因治疗的一个新靶点。

Objective This study was to evaluate the roles of glucose regulated protein - 78 （GRP78） in invasion and metastasis of breast cancer cell line MDA - MB - 231. Methods Reverse transcriptase - polymerase chain reaction （RT- PCR） products of GRP78cDNA fragments were cloned into plasmid pEGFP- C1 in antisense direction. The recombinant pEGFP- C1/GRP78 was transfected into MDA- MB -231 cells. GRP78 mRNA and protein expression, and cell invasion ability were detected by RT- PCR, Western blot, wound healing assay and invasion ability test, respectively. Results Antisense GRP78 eukaryotic expression vector pEGFP - C1/GRP78 was successfully constructed. After 12h and 24h of transfection with pEGFP - C1/GRP78, the expression of GRP78mRNA and protein in pEGFP - C1/GRP78 - transfected MDA - MB - 231 cells was significantly lower than that of non - transfected cells. The tumor cell motility and invasive potential were weakened in pEGFP - C1/GRP78 - transfected cells. Conclusion GRP78 is one of the metastasis genes and enhances breast cancer cell metastasis and may be useful in gene therapy for breast carcinoma.