Di Bisceglie A M, Dusheiko G M, Paterson A C, Alexander J, Shouval D, Lee C S, Beasley R P, Kew M C
Br J Cancer. 1986 Nov;54(5):779-85. doi: 10.1038/bjc.1986.240.
Alpha-foetoprotein (AFP) synthesis, although repressed in normal adults, is increased in the majority of patients with hepatocellular carcinoma (HCC). We have investigated whether active transcription of the AFP gene may explain raised serum AFP concentrations in patients with HCC and hepatoblastoma by assaying human tumour and non-neoplastic tissue by molecular hybridization for the presence of mRNA encoding AFP. Ten operative HCC and six autopsy HCC specimens, two HCC cell lines, and one hepatoblastoma specimen were examined. Total cellular RNA and poly-(A)+ RNA were extracted and AFP mRNA sequences sought by dot-blot and Northern blot hybridisation to a human cDNA AFP probe. Cellular AFP was localised by avidin-biotin staining. AFP mRNA was detected in 8/10 operative specimens, as well as PLC/PRF/5 nude mouse tumours. Weaker hybridization was detected in 4/6 autopsy specimens. Signals of comparable intensity to that in operative tumours were detected in non-neoplastic tissue of 6 patients. AFP mRNA from nude mouse tumours migrated as a 20S discrete band on agarose gel electrophoresis, whereas a more complex hybridization pattern was evident in human tumours. Positive cytoplasmic immuno-staining for AFP was observed in 4 tumours and 2 corresponding non-neoplastic specimens and in a HCC cell line. In non-neoplastic liver, AFP was localised in cells that appeared dysplastic. Thus steady-state levels of AFP mRNA are detectable in human HCC tissue and surrounding non-neoplastic liver. These findings may prove pertinent to an understanding of the genetic expression of AFP in malignant hepatocytes, and the sequence of events leading to uncontrolled cellular proliferation.
甲胎蛋白（AFP）的合成在正常成年人中受到抑制，但在大多数肝细胞癌（HCC）患者中会增加。我们通过分子杂交检测编码AFP的mRNA的存在，以研究AFP基因的活性转录是否可以解释HCC和肝母细胞瘤患者血清AFP浓度升高的原因，检测对象为人肿瘤组织和非肿瘤组织。检查了10个手术切除的HCC标本、6个尸检HCC标本、2个HCC细胞系和1个肝母细胞瘤标本。提取总细胞RNA和聚腺苷酸（poly - A）+ RNA，通过斑点印迹和Northern印迹杂交，使用人AFP cDNA探针寻找AFP mRNA序列。通过抗生物素蛋白-生物素染色对细胞AFP进行定位。在10个手术标本中的8个以及PLC/PRF/5裸鼠肿瘤中检测到AFP mRNA。在6个尸检标本中的4个检测到较弱的杂交信号。在6例患者的非肿瘤组织中检测到与手术肿瘤中强度相当的信号。裸鼠肿瘤中的AFP mRNA在琼脂糖凝胶电泳上迁移为20S离散条带，而在人类肿瘤中杂交模式更为复杂。在4个肿瘤、2个相应的非肿瘤标本以及1个HCC细胞系中观察到AFP的阳性细胞质免疫染色。在非肿瘤肝脏中，AFP定位于出现发育异常的细胞中。因此，在人类HCC组织和周围的非肿瘤肝脏中可检测到AFP mRNA的稳态水平。这些发现可能有助于理解AFP在恶性肝细胞中的基因表达以及导致细胞不受控制增殖的事件顺序。
