摘要
背景与目的目前肺癌治疗中存在无早期特异性诊断指标的难题。本研究旨在获得hnRNPA2/B1cDNA序列,研究其在肺癌组织中的表达情况,为肺癌早期诊断治疗提供实验依据。方法从培养的A549肺腺癌细胞中提取总RNA进行逆转录反应,经PCR扩增目的片段,将其克隆于PUCm-T质粒上,并经酶切电泳、DNA测序鉴定分析。采用原位杂交方法,利用特异的cDNA探针,检测41例肺癌、13例非肺癌组织中hnRNPA2/B1的表达情况。结果从培养的A549肺腺癌细胞提取的总RNA中扩增到目的片段,并经测序证实该片段为hnRNPA2/B1编码序列。原位杂交显示肺癌组hnRNPA2/B1阳性率为87.8%(36/41),显著高于非肺癌组23.1%(3/13)(P<0.01)。其在胞浆、胞核均有表达,主要在胞浆表达。四种类型肺癌组织均显示hnRNPA2/B1阳性染色,其中鳞癌细胞染色阳性率为91.3%(21/23),似高于腺癌细胞78.6%(11/14),但两组间无显著性差异(P>0.05)。结论hnRNPA2/B1可以被成功克隆。初步证实hnRNPA2/B1在肺癌组织中呈高表达,但与肺癌组织类型无明显关系。
Background and objective There is no any specific dynamic criterion in early diagnosis of lung cancer yet. The aim of this study is to obtain the cDNA sequence of hnRNP A2/B1 and then determine its expression in patients with lung cancer, and to provide experimental data for the early diagnosis and treatment of lung cancer. Methods Total RNA was isolated from A549 cells and RT PCR was performed. The fragment was cloned into PUCm T plasmids and further sequenced, hnRNP A2/B1 expression was detected in 41 patients with lung cancer and 13 patients with carcinoma free diseases by in situ hybridization. Results The fragment was identified byDNA sequencing. The positive rate of hnRNP A2/B1 was 87.8% （36/41） in lung cancer tissues, which was significantly higher than that in control group （23. 1% , 3/13） （P〈0.01）. hnRNP A2/B1 was localized in the cytoplasm and/or nucleus, but mainly in the cytoplasm. Four subtypes of lung cancer all showed positive staining of hnRNP A2/B1 protein. The positive rate of hnRNP A2/B1 was 91.3% （21/23） in squamous cell carcinoma, and 78.6% （11/14） inadenocarcinoma （P〉0.05）. Conclusion hnRNP A2/B1 has been cloned successfully. It can highly express in lung cancer tissues, but it does not correlate with histological classification of lung cancer.
出处
《中国肺癌杂志》
CAS
2005年第4期266-269,共4页
Chinese Journal of Lung Cancer
