CDKN2/p16^(INK4a)基因克隆、探针制备及其在肺癌基因诊断中的应用

CDKN2/p16 gene cloning and its probe preparation and application in lung cancer

目的 克隆p16基因 ,制备探针 ,研究p16基因在人肺癌中的变化。 方法 制备总RNA ,逆转录PCR ,构建p16质粒 ,进行基因组DNASouthern杂交 ,检测了 46例人肺癌组织标本和 3例正常肺组织、6例癌旁组织、3例肺癌转移的淋巴结标本中p16基因的情况。 结果 EcoRI酶切进行基因组DNASouthern杂交 ,3例正常组织和 6例癌旁组织标本在 4 3kb处呈阳性杂交带 ,46例肺癌标本有 8例在 4 3kb处缺失 ,缺失率为 17 4%。 结论 p16基因缺失可能在肺癌的发生、发展中起一定作用。

Abstracts[WT5”BZ] To clone CDKN2/p16 INK4a gene, prepare its probe, and to study the change of CDKN2/p16 INK4a gene in lung cancers. [WT5”HZ]Methods[WT5”BZ] Total RNA of normal lung tissue was extracted, CDKN2/p16 INK4a gene cDNA synthesized, and CDKN2/p16 INK4a gene recombinant vector,constructed. Southern blot was used to study CDKN2/p16 INK4a gene in 46 cases of lung cancers, 3 cases of normal lung tissues, 6 cases of lung tissues near cancer, and 3 cases of lymph nodes with lung cancer metastasis. [WT5”HZ]Results[WT5”BZ] Cloned CDKN2/p16 INK4a cDNA was proved by enzyme digestion and sequencing. Southern blot showed 4.3 kb band in normal lung tissues and lung tissues near cancers, and deletion of CDKN2/p16 INK4a gene in cancer tissues and lymph nodes with lung cancer metastasis, with a deletion rate of 17.4%(8/46). [WT5”HZ]Conclusion[WT5”BZ] CDKN2/p16 INK4a gene may play a role to some extent in progression of lung cancers. [WT5”HZ]