反义P^(53)基因和mdm_2基因对裸鼠致瘤性的影响

The Effect of Anti-sense P^(53) and mdm_2 Gene Transferred into GLC-82 Cells on Growth and Tumorigencity of Nude Mice

目的：研究反义P53基因和mdm2基因对裸鼠致癌性的影响，为肺腺癌基因治疗提供新思路和新方法。方法：将含有反义P53基因（PDOR-FP53）和mdm2基因（PDOR-mdm2）及空载体（PDOR-neo）的逆转录表达载体通过脂质体介导分别转染GLC-82细胞，经G418筛选得到抗性克隆。采用流式细胞仪测定细胞周期，软琼脂培养集落形成试验观察集落形成。取2X106／ml细胞悬液分别接种子裸鼠左后肢股部皮下，并与接种GLC-82亲本细胞和接种生理盐水空白对照组比较。结果：转染PDOR-FP53／mdm2的细胞周期（G172．9％，S17．1％，G210．0％）与对照空载体（G141．8％，S53．6％，G245％）比较，G1期细胞峰增高，S期细胞峰降低。软琼脂集落形成试验显示GLC-82亲本细胞和PDOR-neo集落形成早而多，PDOR-FP53／mdm2比对照组晚两周才出现少全集落。裸鼠体内致癌试验表明转染PDOR-FP53mdm2基因的细胞接种裸鼠后出瘤时间晚，生长缓慢且瘤体小。相反GLC-82亲本和转染PDOR-neo的细胞出瘤时间早，生长速度快且瘤体大。结论：反义P53基因和mdm2基因对裸鼠肺腺癌成癌性有双重抑制作用。

Objective Inhibition effect of anti-sense P53 and mdm, gene on tumorigencity of nude mice. Methods Through lipofectanine mediated DNA transfection, PDoR-FP53 , PDOR -mdm2,PDOR-neo introduced into GLC- 82 cells and assessed G418-resitant clonal growth. The expression PDOR-FP53/mdm2 of flow cytome try test and colony-foeming test. 2 X 106/ml cells inoculated into each subcutaneous in nude mice were given and compared with saline or vectpr plasmid administration groups. Results The results of FCM showed that the peak of PDOR-FP53/mdm2 transferred GLC-82 cell was obviously higher in G1 Phase and Lower in S phase (G172. 9%, S 17. 15%, G210. 0%), which was indicative of the inhibition of cell multiplication and the block of DNA synthesis. Colony forming test in soft agar culture and tumorigencity test in nude mice showed that colony -forming rate and tumorigencity were decreased in PDOR - FP53/mdm: transferred GLC- 82 cells. These results indicated that transfection of PDOR-FP53/mdm2 could inhibit the growth of the GLC- 82 cells and reduce the tumorigencity. Conclusion Inhibition effect of anti-sense P53 and mdm2 gene on glandular lung cancer cell tumorigencity of nude mice.