Yago K, Zenita K, Ginya H, Sawada M, Ohmori K, Okuma M, Kannagi R, Lowe J B
Laboratory of Experimental Pathology, Aichi Cancer Center, Nagoya, Japan.
Cancer Res. 1993 Nov 15;53(22):5559-65.
Transcripts of alpha-(1,3)-fucosyltransferases in human epithelial cancer and leukemia cell lines were analyzed by Northern blotting and reverse transcriptase mediated-polymerase chain reaction using specific probes and primers which can discriminate between the transcripts derived from the four alpha-(1,3)-fucosyltransferase genes Fuc-TIII, IV, V, and VI. Flow cytometric analysis of the sialyl Le(x) and sialyl Le(a) antigens was also performed on the same cell lines. The sialyl Le(x) antigen was expressed on 14 of 15 epithelial cancer cell lines, and the sialyl Le(a) antigen was detected on 8 of them. The message of Fuc-TIII was detected in most of the epithelial cancer cell lines (14 of 15), which correlated with the surface expression of these carbohydrate determinants. In addition, the messages of Fuc-TIV and Fuc-TVI were detected in most epithelial cancer cell lines, while the message of Fuc-TV was undetectable in most of them. On the other hand, all leukemia cell lines were positively stained for sialyl Le(x), but none of them was stained for sialyl Le(a) in flow cytometry. The messages of Fuc-TIV++ were detected in all leukemia cell lines tested. Small quantities of Fuc-TIII, V, and/or VI messages were also detected in some leukemia cell lines in reverse transcriptase mediated-polymerase chain reaction analysis. These studies indicate that alpha-(1,3)-fucosyltransferase activities in epithelial cancer and leukemia cell lines are mixtures of multiple molecular species of alpha-(1,3)-fucosyltransferases. It is natural that epithelial cancer cells contain a significant amount of Fuc-TIII mRNA and leukemia cell lines contain Fuc-TIV mRNA, since their normal counterparts, normal epithelial cells and leukocytes, respectively, are known to contain these fucosyltransferases. The unexpectedly frequent occurrence of Fuc-TIV mRNA in epithelial cancer cell lines may be related to their retro-differentiation associated with tumorigenesis. Another unexpected finding was a weak but significant expression of the alpha-(1,3)-fucosyltransferases Fuc-TIII, V, and/or VI in leukemia cell lines detected by reverse transcriptase mediated-polymerase chain reaction analysis. Since these enzymes are known to be capable of synthesizing the sialyl Le(x) determinant, this finding implies a possibility that some of them may be involved in the synthesis of sialyl Le(x) in leukemia cells.
利用特异性探针和引物,通过Northern印迹法以及逆转录酶介导的聚合酶链反应,对人上皮癌细胞系和白血病细胞系中的α-(1,3)-岩藻糖基转移酶转录本进行了分析,这些探针和引物能够区分源自4个α-(1,3)-岩藻糖基转移酶基因Fuc-TIII、IV、V和VI的转录本。同时,对相同的细胞系进行了唾液酸化Le(x)和唾液酸化Le(a)抗原的流式细胞术分析。15个上皮癌细胞系中有14个表达唾液酸化Le(x)抗原,其中8个检测到唾液酸化Le(a)抗原。在大多数上皮癌细胞系(15个中的14个)中检测到Fuc-TIII的信息,这与这些碳水化合物决定簇的表面表达相关。此外,在大多数上皮癌细胞系中检测到Fuc-TIV和Fuc-TVI的信息,而在大多数细胞系中未检测到Fuc-TV的信息。另一方面,所有白血病细胞系在流式细胞术中唾液酸化Le(x)均呈阳性染色,但唾液酸化Le(a)均未染色。在所有检测的白血病细胞系中均检测到Fuc-TIV++的信息。逆转录酶介导的聚合酶链反应分析还在一些白血病细胞系中检测到少量的Fuc-TIII、V和/或VI信息。这些研究表明,上皮癌细胞系和白血病细胞系中的α-(1,3)-岩藻糖基转移酶活性是多种α-(1,3)-岩藻糖基转移酶分子种类的混合物。上皮癌细胞含有大量Fuc-TIII mRNA以及白血病细胞系含有Fuc-TIV mRNA是很自然的,因为它们的正常对应物,即正常上皮细胞和白细胞,已知分别含有这些岩藻糖基转移酶。上皮癌细胞系中Fuc-TIV mRNA意外频繁出现可能与它们与肿瘤发生相关的逆分化有关。另一个意外发现是,逆转录酶介导的聚合酶链反应分析在白血病细胞系中检测到α-(1,3)-岩藻糖基转移酶Fuc-TIII、V和/或VI的弱但显著的表达。由于已知这些酶能够合成唾液酸化Le(x)决定簇,这一发现意味着其中一些酶可能参与白血病细胞中唾液酸化Le(x)合成的可能性。