摘要
目的研究逆转录病毒介导的ｐ１６基因对人卵巢癌细胞系ＣＡＯＶ３生长与致瘤性的抑制作用。方法应用脂质体介导法，将外源性野生型ｐ１６基因转染不表达ｐ１６的人卵巢癌细胞系ＣＡＯＶ３，对转染后细胞ＤＮＡ、ＲＮＡ和蛋白进行分析并观察其生物学行为变化。结果外源性野生型ｐ１６基因已整合于细胞并获稳定表达，表达有外源ｐ１６基因的细胞生长速度、集落形成率及裸鼠致瘤性均有部分抑制。细胞周期分析可见Ｇ１期增加，Ｓ期下降。电镜下超微结构出现生长抑制特征。结论ｐ１６基因可明显抑制癌细胞生长，在卵巢癌发生、发展的过程中起重要作用。
Objective To study the inhibitory effects of retrovirusmediated p16 gene on human ovarian cancer cell line CAOV3. Methods Recombinant eukaryotic expression vector pDORp16 containing exogenous human wtp16 cDNA and vector with neomycin resistance gene only were introduced by lipofectaminemediated gene transfection into CAOV3 cell line which does not express p16 endogenously.By using revertpolymerase chain reaction amplification,mRNA in situ hybridization and immunocytochemistry,the clones obtained were detected for their efficiency of transfection and effects of vector expression and observed for their biologic behavior. Results Exogenous wtp16 had successfully been transferred into CAOV3 cells and obtained permanent expression. The growth rate of these transfected CAOV3 cells in regular medium and soft agar was inhibited, and the tumorigenicity in nude mice showed that two in four mice failed to form tumor and the others suffered from tumor later than contrast group by 7 to 14 days. The percentage of phase G1 cells increased and that of phase S cells decreased by analysing cell cycle. The ultrastructural changes of the cells were observed under electron microscope, revealing necrosis and growth retardation. Conclusions p16 gene played an important rolein generation and development of ovarian carcinoma. This study might provide the experimental evidence for the gene therapy in human ovarian cancer.
出处
《中华妇产科杂志》
CAS
CSCD
北大核心
1999年第5期304-307,I007,共4页
Chinese Journal of Obstetrics and Gynecology
基金
辽宁省自然科学基金
