Yu C J, Yang P C, Shew J Y, Hong T M, Yang S C, Lee Y C, Lee L N, Luh K T, Wu C W
Department of Internal Medicine, National Taiwan University Hospital, Taipei, Republic of China.
Oncology. 1996 Mar-Apr;53(2):118-26. doi: 10.1159/000227547.
The expression pattern of mucin genes was studied in 7 lung adenocarcinoma cell lines (CL1, CL2, CL3, NCL2, PC9, PC13, PC14) and 12 lung adenocarcinoma tissues. CL1 and PC13 are poorly differentiated cell lines with low mucin glycoprotein production. The other 5 cell lines are well differentiated and produce a higher amount of mucins. Total RNA was extracted from these cell lines. Northern blot analysis was performed by hybridization with specific antisense oligonucleotide probes recognizing mucin-specific tandem repeats of 4 mucin genes (MUC1, MUC2, MUC3, MUC4). RT-PCR was carried out to amplify the 3' and 5' nonrepetitive coding regions of MUC1 and the 5' nonrepetitive coding region of MUC2. All these cell lines expressed MUC1, MUC2, and MUC4 mRNA but in variable mounts. The poorly differentiated cell lines (CL1 and PC13) had a relatively low level of expression of MUC1, MUC2, MUC3 and MUC4. RT-PCR, with primers amplifying the MUC1 nonrepetitive coding region 5' end, 293 bp, and the 3' end, 522 bp, as well as the MUC2 nonrepetitive 5' coding region, 308 bp, revealed the presence of MUC1 and MUC2 mRNA in all the cell lines. Sequence analysis of the PCR products were very homologous, similar to previously published MUC1 and MUC2 cDNA sequences. The expression pattern of mucin genes is consistent with that of mucin glycoproteins as studied using biochemical and immunological methods. Northern blotting and RT-PCR analysis in 12 lung adenocarcinoma tissues with various grades of differentiation (6 poorly differentiated adenocarcinomas and 6 moderately to well-differentiated adenocarcinomas) showed heterogeneous expression of the 4 mucin genes in tissues without clear correlation with the differentiation grade. Therefore the clinical implications of the differential expression of the mucin genes need further investigation.
在7种肺腺癌细胞系(CL1、CL2、CL3、NCL2、PC9、PC13、PC14)和12例肺腺癌组织中研究了黏蛋白基因的表达模式。CL1和PC13是低分化细胞系,黏蛋白糖蛋白产生量低。其他5种细胞系分化良好,产生的黏蛋白量较高。从这些细胞系中提取总RNA。通过与识别4种黏蛋白基因(MUC1、MUC2、MUC3、MUC4)黏蛋白特异性串联重复序列的特异性反义寡核苷酸探针杂交进行Northern印迹分析。进行RT-PCR以扩增MUC1的3'和5'非重复编码区以及MUC2的5'非重复编码区。所有这些细胞系均表达MUC1、MUC2和MUC4 mRNA,但表达量各不相同。低分化细胞系(CL1和PC13)中MUC1、MUC2、MUC3和MUC4的表达水平相对较低。用扩增MUC1非重复编码区5'端(293 bp)、3'端(522 bp)以及MUC2非重复5'编码区(308 bp)的引物进行RT-PCR,结果显示所有细胞系中均存在MUC1和MUC2 mRNA。PCR产物的序列分析显示其具有高度同源性,与先前发表的MUC1和MUC2 cDNA序列相似。黏蛋白基因的表达模式与使用生化和免疫方法研究的黏蛋白糖蛋白的表达模式一致。对12例不同分化程度的肺腺癌组织(6例低分化腺癌和6例中分化至高分化腺癌)进行Northern印迹和RT-PCR分析,结果显示4种黏蛋白基因在组织中的表达存在异质性,与分化程度无明显相关性。因此,黏蛋白基因差异表达的临床意义需要进一步研究。