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背景与目的 :探讨新抑癌基因KAI1与全反式维甲酸(all-trans-retinoicacid,ATRA)对小细胞肺癌NCI-H446细胞株抑制增殖和诱导分化的作用。材料与方法 :用脂质体介导的基因转染方法 ,借助质粒表达载体(PCMV -NEO -XhoI),将抑癌基因KAI1转入小细胞肺癌NCI-H446细胞中 ,经G418筛选 ,获得稳定表达的细胞克隆。用10-6 mol/LATRA作用于转染及未转染KAI1基因的小细胞肺癌NCI-H446细胞株,集落形成率检测细胞体外增殖能力 ,流式细胞仪进行细胞周期和凋亡分析,间接免疫荧光染色结合流式细胞仪检测转染前后细胞CD82蛋白的表达。免疫组化测定MYC的表达 ,放射免疫测定LN(层连蛋白)表达。结果 :ATRA处理脂质体 -KAI1基因转染的小细胞肺癌细胞CD82表达降低 ,细胞增殖能力下降 ,凋亡增加 ,更多的细胞被阻止于G1/G0 期 ,MYC及LN表达下降。结论 :抑癌基因KAI1与ATRA对抑制小细胞肺癌NCI-H446细胞株的增殖和促分化有协同作用。
BACKGROUND & AIM: To investigate the inhibitory effect of KAI1 and all-trans-retinoic acid (ATRA) on the proliferation and differentiation of small cell lung cancer cell line NCI-H446. MATERIALS AND METHODS: The suppressor gene KAI1 was transfected into small cell lung cancer NCI-H446 cells with lipofectamine mediated gene transfection method using plasmid expression vector (PCMV-NEO-XhoI), and was screened by G418 to obtain stable Expressed cell clone. The small cell lung cancer NCI-H446 cells were transfected with or without untransfected KAI1 with 10-6 mol / L ATRA. The proliferation of NCI-H446 cells was detected by colony formation assay. Cell cycle and apoptosis were analyzed by flow cytometry. Fluorescent staining combined with flow cytometry before and after transfection CD82 protein expression. The expression of MYC was detected by immunohistochemistry and LN (laminin) expression was measured by radioimmunoassay. Results: ATRA treatment of liposome-KAI1 transfected small cell lung cancer cells CD82 expression decreased, cell proliferation decreased, increased apoptosis, more cells were blocked in G1 / G0, MYC and LN decreased. Conclusion: KAI1 and ATRA have synergistic effects on the proliferation and differentiation of small cell lung cancer cell line NCI-H446.