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目的对肺癌转移相关基因C_1,L_1,L_2片断(已知序列)进行验证,并克隆其基因全长。方法应用PCR及快速扩增cDNA末端(RACE)技术,在小细胞肺癌细胞株(NCI-H446细胞)和肺腺癌细胞株(95-D细胞)中对肺癌转移相关基因C_1、L_1、L_2片断进行验证并克隆出其全长序列,登录基因库。结果在NCI-H446细胞和95-D细胞中未得到C_1和L_1目的片段,只得到L_2目的片段。从这两种细胞株中均钓取到L_2基因的3′端部分未知序列1 169 bp;从NCI-H446细胞、95-D细胞中分别钓取得到L_2基因的5′端未知序列178bp和145bp。L_2基因序列与已知的ABCE1基因序列同源性为100%。结论L_2基因即为已知ABCE1基因,可能参与肺癌转移的相关调节。
Objective To verify the C_1, L_1 and L_2 fragments (known sequences) of lung cancer metastasis-related genes and clone their full-length genes. Methods The gene fragments of C_1, L_1 and L_2 in lung cancer were detected by PCR and rapid amplification of cDNA ends (RACE) in small cell lung cancer cell line (NCI-H446) and lung adenocarcinoma cell line (95-D cell) Validate and clone their full-length sequence and log in to the GenBank. Results The target fragment of C_1 and L_1 was not found in NCI-H446 cells and 95-D cells, and only the target fragment of L_2 was obtained. From these two cell lines, 1 169 bp of the 3 ’end of unknown L_2 gene was captured; from the NCI-H446 cells and 95-D cells, the 5’ end unknown sequence of L_2 gene was found to be 178 bp and 145 bp, respectively . The sequence homology between L_2 gene and known ABCE1 gene is 100%. Conclusion L_2 gene is a known ABCE1 gene, which may be involved in the regulation of lung cancer metastasis.