2018 年第 3 期 第 13 卷

脂滴包被蛋白基因反义RNA对人脂肪肉瘤细胞SW872增殖活性的影响

Effect of perilipin gene antisense RNA on proliferation of human liposarcoma cell line SW872

作者：孟令新厉兵城郑玉秀丁兆军徐美玲孙树艳孟芹杨万才

英文作者：

单位：276826山东省日照市人民医院肿瘤科(孟令新、厉兵城、郑玉秀、丁兆军、徐美玲、孟芹)，病理科(孙树艳)；272067济宁医学院精准医学研究院(杨万才)

英文单位：

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• 摘要：

• 目的    观察脂滴包被蛋白(perilipin)基因反义RNA对人脂肪肉瘤细胞SW872增殖能力的影响。方法    将含perilipin基因反义cDNA序列的pcDNA3.1-perilipin(-)质粒及pcDNA3.1空载体质粒转染入人脂肪肉瘤SW872细胞，分别命名为SW872-A组和SW872-E组；未转染的瘤细胞为空白对照(SW872-wt组)。用逆转录聚合酶链反应法和链霉亲和素-生物素-过氧化物酶复合物免疫组织化学方法检测perilipin反义基因及perilipin蛋白表达。用噻唑蓝还原法检测SW872细胞增殖活性；用流式细胞术检测细胞凋亡和增殖周期分布。将转基因SW872脂肪肉瘤细胞接种于balb/c裸鼠后肢皮下，4周后观察成瘤情况。结果    SW872-E组和SW872-wt组细胞perilipin mRNA表达明显高于SW872-A组；SW872-A组perilipin阳性细胞数较另2组明显减少。SW872-A组细胞在第1～6天增殖能力逐渐增加，与SW872-E组和SW872-wt组比较，差异均有统计学意义(均P＜0.05)。SW872-A组S期和G2/M期细胞比例明显大于SW872-E组和SW872-wt组［(39±4)%比(19±5)%、(20±3)%，(40±4)%比(20±3)%、(21±3)%］，差异均有统计学意义(均P＜0.05)。SW872-A组裸鼠皮下移植瘤的成瘤时间明显短于SW872-E组和SW872-wt组［(5.3±2.6)d比(10.3±2.2)、(11.2±2.7)d］；接种后28 d，SW872-A组瘤体重量明显大于SW872-E组和SW872-wt组［(3.87±0.46)g比(1.62±0.73)、(1.83±0.24)g］，差异均有统计学意义(均P＜0.05)。结论    perilipin基因反义RNA可抑制SW872脂肪肉瘤细胞中脂滴包被蛋白的表达，促进裸鼠体内肿瘤生长。

• Objective    To investigate the effect of perilipin gene antisense RNA on proliferation of human liposarcoma cell line SW872. Methods    Perilipin gene antisense cDNA plasmids［pcDNA3.1-perilipin(-)］ were transfected into human liposarcoma cell line SW872 as SW872-A group; empty plasmids were transfected into SW872 cells as SW872-E group; blank SW872 cells were SW872-wt group. Expressions of antisense perilipin gene and perilipin protein were tested by reverse transcription polymerase chain reaction and strept avidin biotin-peroxidase complex method. Proliferative activity of SW872 cells was tested by MTT assay. Apoptosis and proliferating cycle of SW872 cells were tested by flow cytometry. Transgenic SW872 cells were subcutaneously transplanted at posterior limbs of balb/c nude mice; liposarcoma growth was observed in 4 weeks. Results    Expression of perilipin mRNA in SW872-E group and SW872-wt group was significantly higher than that in SW872-A group. Perilipin positive cell number in SW872-A group was significantly less than that in SW872-E group and SW872-wt group. Proliferative activity of SW872 cells in SW872-A group was significantly higher than that in SW872-E group and SW872-wt group on the 1st-6th days after transfection(P＜0.05). Proportions of S phase and G2/M phase cells in SW872-A group were significantly higher than those in SW872-E group and SW872-wt group［(39±4)% vs (19±5)%, (20±3)%; (40±4)% vs (20±3)%, (21±3)%］(P＜0.05). Subcutaneous tumor formation time in SW872-A group was significantly shorter than that in SW872-E group and SW872-wt group［(5.3±2.6)d vs (10.3±2.2), (11.2±2.7)d］; tumor weight 28 d after transplantion in SW872-A group was significantly more than that in SW872-E group and SW872-wt group［(3.87±0.46)g vs (1.62±0.73), (1.83±0.24)g］(P＜0.05). Conclusion    Perilipin gene antisense RNA can inhibit perilipin expression in SW872 liposarcoma cells and promote tumor growth in nude mice.