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CA19-9表位——一种可能的MUC-1/Y蛋白标志物。

CA19-9 epitope a possible marker for MUC-1/Y protein.

作者信息

Akagi J, Takai E, Tamori Y, Nakagawa K, Ogawa M

机构信息

National Kumamoto-Minami Hospital, Kumamoto 869-0593, Japan.

出版信息

Int J Oncol. 2001 May;18(5):1085-91. doi: 10.3892/ijo.18.5.1085.

DOI:10.3892/ijo.18.5.1085
PMID:11295060
Abstract

It has been reported that MUC-1 molecules devoid of the tandem repeat region (MUC-1/Y) are detected preferentially in carcinoma cells and are associated with their progression. However, its clinical significance is still unknown. We constructed a mouse colon adenocarcinoma cell line (MC-38) transduced with either MUC-1/Y cDNA defecting the tandem repeat region (Y-MC-38) or MUC-1/R cDNA containing ten tandem repeats (R-MC-38). RT-PCR of mRNAs derived from Y-MC-38 cells using the specific primers to MUC-1/Y mRNAs, proved the existence of 600 bp RT-PCR products generated only from MUC-1/Y mRNAs. DF3 and CA19-9 epitopes out of the MUC-1-related tumor-associated antigens, have been reported to be involved in the prognosis of cancer patients. We examined the expression of DF3 and CA19-9 epitopes on Y-MC-38 and R-MC-38 cells. Fluorescence-activated cell sorting (FACS) analysis of R-MC-38 and Y-MC-38 cells using two monoclonal antibodies against DF3 (mAb DF3) and CA19-9 (mAb CA19-9) epitopes revealed that R-MC-38 cells expressed DF3 but not CA19-9 [DF3(+)CA19-9(-)], while Y-MC-38 cells expressed CA19-9 but not DF3 [DF3(-)CA19-9(+)]. On Western blot, a 40 kDa protein product was recognized by mAb CA19-9 but not by mAb DF3 on cell lysates of Y-MC-38 cells, whereas a 70 kDa protein product was recognized by mAb DF3 but not by mAb CA19-9 on the cell lysates of R-MC-38. Further, we analyzed the expression of MUC-1/Y mRNAs by RT-PCR on various human cancer cell lines: the gastric cancer cell line AZ521, the pancreatic cancer cell lines PANC-1 and Capan-1, the gall bladder cell line GBK-1, the breast cancer cell line MCF-7, and the colon cancer cell lines HT-29 and Colo205. HT-29 and Capan-1 cells producing the 600 bp RT-PCR product, were positive for mAb CA19-9. These results demonstrate that CA19-9 epitope is produced only on MUC-1/Y core protein, suggesting that CA19-9 epitope may be a specific marker for MUC-1/Y protein.

摘要

据报道,缺乏串联重复区域的MUC-1分子(MUC-1/Y)在癌细胞中被优先检测到,并与其进展相关。然而,其临床意义仍不清楚。我们构建了一种小鼠结肠腺癌细胞系(MC-38),该细胞系转导了缺失串联重复区域的MUC-1/Y cDNA(Y-MC-38)或含有十个串联重复的MUC-1/R cDNA(R-MC-38)。使用针对MUC-1/Y mRNA的特异性引物对来自Y-MC-38细胞的mRNA进行RT-PCR,证明仅从MUC-1/Y mRNA产生了600 bp的RT-PCR产物。据报道,MUC-1相关肿瘤相关抗原中的DF3和CA19-9表位与癌症患者的预后有关。我们检测了Y-MC-38和R-MC-38细胞上DF3和CA19-9表位的表达。使用两种针对DF3(单克隆抗体DF3)和CA19-9(单克隆抗体CA19-9)表位的单克隆抗体对R-MC-38和Y-MC-38细胞进行荧光激活细胞分选(FACS)分析,结果显示R-MC-38细胞表达DF3但不表达CA19-9 [DF3(+)CA19-9(-)],而Y-MC-38细胞表达CA19-9但不表达DF3 [DF3(-)CA19-9(+)]。在蛋白质印迹法中,Y-MC-38细胞裂解物上的一种40 kDa蛋白质产物被单克隆抗体CA19-9识别,但不被单克隆抗体DF3识别,而R-MC-38细胞裂解物上的一种70 kDa蛋白质产物被单克隆抗体DF3识别,但不被单克隆抗体CA19-9识别。此外,我们通过RT-PCR分析了多种人类癌细胞系中MUC-1/Y mRNA的表达:胃癌细胞系AZ521、胰腺癌细胞系PANC-1和Capan-1、胆囊细胞系GBK-1、乳腺癌细胞系MCF-7以及结肠癌细胞系HT-29和Colo205。产生600 bp RT-PCR产物的HT-29和Capan-1细胞对单克隆抗体CA19-9呈阳性。这些结果表明,CA19-9表位仅在MUC-1/Y核心蛋白上产生,提示CA19-9表位可能是MUC-1/Y蛋白的特异性标志物。

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