摘要
目的:探讨新基因CIAPIN1在人肺癌和癌旁正常组织中的表达差异及该基因导入对人小细胞肺癌NCI-H446细胞生长的调控作用。方法:采用免疫组化方法检测51例石蜡包埋的肺癌及癌旁正常肺组织中CIAPIN1蛋白的表达;构建腺病毒载体介导CIAPIN1基因(Ad-CIAPIN1),转染到自身CIAPIN1低表达的人小细胞肺癌细胞系NCI-H446,Westernblot检测目的蛋白的表达;台盼蓝染色计数活细胞数,绘制细胞生长曲线;流式细胞术(FCM)分析细胞周期及细胞凋亡的变化。结果:癌旁正常肺组织中CIAPIN1基因蛋白阳性表达率(100%)明显高于肺癌组织中表达率(39.2%,P<0.05);与对照组相比,腺病毒介导的CIAPIN1实验组能迅速抑制NCI-H446细胞的生长(P<0.01),诱导细胞发生凋亡,并出现明显的G1/S期阻滞现象。结论:肺癌组织中CIAPIN1基因表达下调可能与肺癌发生密切相关,Ad-CIAPIN1转染NCI-H446细胞能明显抑制肿瘤细胞的生长,提示CIAPIN1基因可能是一个新的抑癌相关基因。
AIM: To explore CIAPIN1 gene expression in lung carcinoma tissues and its regulatory in the growth of NCI-H446. METHODS: Fifty-one formalin-fixed paraffin-embedded specimens of primary lung cancer and their noncancerous counterparts were detected by SABC immunohistochemistry method. Adenoviral vector construction was recombined and the gene was transduced into NCI-H446 cells. The expression of CIAPIN1 protein was identified by Western blot. Trypan blue staining was used to count the alive cells and to draw a cellular growth curve. The changes of cell cycle and apoptosis were analyzed by flow cytometry. RESULTS: The positive rate of CIAPIN1 expression in cancer tissues (39.2%) was much lower than that in non-cancerous counterparts( 100%, P 〈 0.05). In Ad-CIAPIN1 group, the growth of transfected NCI-H446 cells in vitro was significantly inhibited. In addition the Ad-CIAPIN1-induced cell apoptosis and a predominant arrest in the G1/S phase ( P 〈 0.01 ) were observed. CONCLUSION: The down-regulation of CIAPIN1 expression in tumor is associated with the development of lung carcinoma. TransducUon of NCI-H446 CIAPIN1-negative cell, with Ad-CIAPIN1 can inhibit cell growth, suggesting that CIAPIN1 can be a new tumor-related suppressor gene.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2008年第5期434-437,共4页
Chinese Journal of Cellular and Molecular Immunology