DIFFERENTIAL DISPLAY AND INTEGRIN-ALPHA-6 MESSENGER-RNA OVEREXPRESSION IN HEPATOCELLULAR-CARCINOMA
DIFFERENTIAL DISPLAY AND INTEGRIN-ALPHA-6 MESSENGER-RNA OVEREXPRESSION IN HEPATOCELLULAR-CARCINOMA
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DOI：
10.1002/hep.1840220518
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发表时间：
1995-11-01
期刊：
影响因子：
13.5
通讯作者：
BARNARD, GF
中科院分区：
文献类型：
--
作者：
BEGUM, NA;MORI, M;BARNARD, GF
我们的目的是分离潜在的重要差异表达的基因产物配对人肝细胞癌（HCC）和正常肝脏样本使用差异信使RNA（mRNA）显示技术。总RNA样品用锚定寡核苷酸引物逆转录，然后用额外的上游随机引物通过聚合酶链反应（PCR）扩增。差异表达的互补DNA（cDNA）产物随后用作北方印迹分析中的探针。一个这样的cDNA产物，存在于肿瘤中，但在正常显示器中不存在，显示出与粘附分子整合素α 6相同。在16对HCC的北方印迹中，7个肿瘤中有类似于5.5kb的整合素α 6 mRNA信号过表达，而正常肝脏中有弱信号。对于整合素α 6 mRNA过表达的患者和无整合素α 6 mRNA过表达的患者：（1）分别在7个肿瘤中观察到III（或IV）级组织学，在9个肿瘤中观察到3个肿瘤（P = .03）;（2）肿瘤复发或死亡（平均随访18个月）分别在7例患者中的6例和8例患者中的3例中观察到，（P = 0.17），用甘油醛S-磷酸脱氢酶作为对照，使用半定量PCR共扩增获得了类似的结果;大约50%的肿瘤具有比其配对的正常细胞更强的整合素α 6带。整合素α 6 mRNA的A和B变体在肿瘤和正常肝脏样品中均可检测到。B变体在肿瘤中比A变体明显8.9倍（n = 10），而在正常肝脏中为3倍（n = 10），这表明整合素α 6的过度表达可能更能反映B变体水平的异常而不是A变体水平的异常。重要的基因，其表达与显着的患者变量可能被隔离的差异显示，基于这个小系列有一个趋势，整合素α 6 mRNA在高级别肝癌过表达，并预测一个趋势，一个较差的结果。
Our aim was to isolate potentially important differentially expressed gene products from paired human hepatocellular carcinoma (HCC) and normal liver samples using the differential messenger RNA (mRNA) display technique. Total RNA samples were reverse transcribed with anchoring oligonucleotide primers and then amplified by the polymerase chain reaction (PCR) with additional upstream random primers. Differentially expressed complementary DNA (cDNA) products were subsequently used as probes in Northern blot analysis. One such cDNA product, present in tumor but absent in normal displays, showed identity with the adhesion molecule integrin alpha 6. In Northern blots of 16 HCC pairs, the similar to 5.5 kb signal of integrin alpha 6 mRNA was overexpressed in seven tumors, with a weak signal in the normal livers, For those patients with versus without integrin alpha 6 mRNA overexpression: (1) grade III (or IV) histology was noted in seven of seven versus three of nine tumors, respectively (P = .03); (2) tumor recurrence or death (at mean follow-up of 18 months) was noted in six of seven versus three of eight patients, respectively (P = .17), Similar results were obtained using semiquantitative PCR co-amplification with glyceraldehyde S-phosphate dehydrogenase as a control; similar to 50% of the tumors had stronger integrin alpha 6 bands than their paired normals. Both A and B variants of integrin alpha 6 mRNA were detectable in the tumor and normal liver samples. The B variant was more pronounced than the A variant by 8.9-fold in the tumors (n = 10) compared with threefold in the normal livers (n = 10), suggesting that the overexpression of integrin alpha 6 may be more reflective of abnormalities of B variant levels than of A variant levels. Important genes whose expression correlates with significant patient variables may be isolated by differential display; based on this small series there is a trend for integrin alpha 6 mRNA to be overexpressed in high-grade HCC and to predict a tendency toward-a poorer outcome.