涎腺多形性腺瘤的倍体分布及与基因治疗的关系

目的:探讨涎腺多形性腺瘤的DNA含量及倍体分布与HSV-tk基因治疗的关系.方法:采用流式细胞仪测定50例涎腺良性多形性腺瘤DNA含量和倍体分布并结合临床资料进行分析;应用腺病毒介导的HSV-tk基因分别转染培养中良性及交界性多形性腺瘤细胞;RT-PCR方法检测基因表达;四唑蓝比色(MTI)法测定HSV-tk系统对二倍体及异倍体肿瘤细胞的杀伤作用.结果:10例正常涎腺组织皆为二倍体;涎腺良性多形性腺瘤异倍体率20.0%;30岁以下年龄,病程在2年以内及瘤体直径＞5cm者异倍体率明显增高(P＜0.05).二倍体肿瘤细胞转染HSV-tk/GCV基因后3、5天,细胞存活率分别为78.3%和37.7%.异倍体肿瘤细胞转染HSV-tk/GCV基因后3、5天,细胞存活率分别为27.6%和27.2%,二者细胞存活率有显著性差异(P＜0.05).结论:1)涎腺多形性腺瘤异倍体检出率与患者的年龄,病程及瘤体直径有密切关系;2)HSV-tk/GCV在治疗后的3、5天对异倍体肿瘤细胞杀伤作用明显高于二倍体肿瘤.

The Relationship between the Gene Therapy and Ploid Distribution in Pleomorphic Adenomas of Salivary Gland

Objective: To discuss the relationship between the DNA content, ploid distribution and the clinical significance and gene therapy for pleomorphic adenoma of salivary gland. Methods: Fifty cases of pleomorphic adenoma and 10 cases of salivary gland were determined by flow cytometry, the HSV-tk gene was transfected into human benign and adjacently malignant pleomorphic adenoma cells of salivary gland respectively using recombinant adenovirus vector. The efficiency of transfection and expression were detected by RT-PCR. The inhibition of cell proliferation was verified by MTT. Results: The aneuploidy rate of pleomorphic adenoma was obviously higher in young patients with large tumor or with the couse of disease shorter than 2 years (P<0.05). The proliferation of pleomorphic adenoma cells infected by HSV-tk gene was inhibited and the survival rates of the diploid cells at day 3 and day 5 were 78.3% and 37.7%, respectively, but the aneuploidy cell survival rate decreased to 27.6% and 27.2% at day 3 and days 5, so the survival rate of the diploid cells was significantly higher (P<0.05). Conclusions: 1) To evaluate the DNA ploidy of pleomorphic adenoma is valuable for diagnosis and treatment. 2) The lethal effect of HSV-tk gene on the aneuploidy cells is obviously much more powerful compared to diploid cells.