肺癌相关基因LSCC-3对肺癌细胞形态特征和生物学活性的作用

目的 观察肺癌相关基因LSCC-3的表达对肺腺癌细胞PAa形态学特征及生物学活性的作用.方法 构建含有LSCC-3基因全长开放阅读框架(ORF)片段之真核表达载体并转染肺癌PAa细胞;显微镜及倒置荧光显微镜下观察LSCC-3基因的表达及其对于肺癌PAa细胞的形态学特征的影响;采用四甲基氮唑盐(MTT)比色法研究LSCC-3基因的转染对于肺癌PAa细胞生物学活性的作用.结果 以pLPS-LSCC-3-3'EGFP真核表达载体成功转染肺癌PAa细胞,显微镜下观察显示细胞质内有大量LSCC-3基因表达的肺癌PAa细胞出现了细胞脱水、核膜皱缩,核小体形成乃至细胞结构破坏等典型的细胞凋亡、破坏迹象.转染LSCC-3基因后第1～5d肺癌PAa细胞的增殖活性(以A490值表示)较对照组细胞受到明显抑制,始终维持在较低水平(0.1157、0.1257、0.1297、0.134、0.145 Vs.0.1165、0.1300、0.1677、0.1935、0.2058);而细胞凋亡率则明显增加,尤其是转染后前3d,实验组细胞的凋亡率几乎呈线性增长,与对照组细胞间差异有统计学意义(0、12.1%、33.2%、37.8%、38.7% Vs 0、9.1%、13.7%、10.3%、13.1%;P<0.01). 结论 LSCC-3基因的表达在体外可以明显地促进肺癌细胞的凋亡,从而抑制肺癌细胞的增殖,其或许可被应用于肺癌的基因或生物治疗.

Effects of LSCC-3 gene on the morphologic characters and biologic activities of lung cancer cells

Objective To study the effects of LSCC-3 gene on the morphologic characters and biologic activities of lung cancer Paa cells by an experimental study in vitro.Methods Lung cancer Paa cells were transfected by pLPS-LSCC-3-3’EGFP.The changes in morphologic characters of lung cancer Paa cells after transfection were observed by fluorescence microscopy.The effects of LSCC-3 gene on the biologic activity and apoptosis rate of lung cancer Paa cells were detected by a MTT method.Results After transfection,the signs of cell apoptosis such as dehydration of cells,appearance of nucleosome,and collapse of nucleolus and cells were found under fluorescence microscopy.Apoptosis rate of Paa cells transfected with LSCC-3 was significantly higher than that of the Paa cells without a transfection within 1-5 days (0,12.1%,33.2%,37.8%,38.7% vs 0,9.1%,13.7%,10.3%,13.1%;P<0.01).Conclusion LSCC-3 gene can promote the apoptosis rate and inhibit the proliferation of lung cancer cells in vitro.LSCC-3 gene can be used in gene therapy or biotherapy of lung cancer.