反义HSP70对人乳腺癌细胞的抑瘤效应

Antitumorigenic Effect of Antisense HSP70 on Human Breast Cancer Cell Line

目的：观察反义热休克蛋白70（heat shock protein 70,HSP70）RNA对人乳腺癌细胞的生物学特性的影响，探讨其抗肿瘤作用。方法：将逆转录病毒真核表达载体介导的反义HSP70重组质粒pX-AHSP70和空载体pLXSN-neo转染至MCF7/Adr人乳腺癌细胞，通过细胞生长曲线和软琼脂集落实验以及裸鼠移植瘤实验观察转染反义HSP70重组质粒细胞的体内外抑瘤作用。结果：建立了稳定表达反义HSP70 RNA的MAp70细胞，与空载体细胞相比，HSP70蛋白的表达下降了48％。细胞群体生长速度明显减慢，软琼脂集落形成率和抑制率分别为6.08％和62.14％，裸鼠皮下接种的平均出瘤时间为15天，最终平均瘤重为108 mg，以上参数与空载体细胞相比，均有显著差异。结论：反义HSP70 RNA能够有效地抑制HSP70表达，进而抑制乳腺癌细胞的体外增殖和体内移植瘤的形成与发展。

Objective：This study was designed to observe the antitumor effect of antisense HSP70 RNA expression on human breast cancer cell line. Methods：An eukaryotic expression vector pX-AHSP70 expressing antisense HSP70 RNA was constructed and transfected into MCF7/Adr human breast cancer cells, which were then selected by antibiotic G418. The expression of HSP70 was detected by Western blot. Furthermore, the tumor repression function of transfected cells were observed in vivo and in vitro through cell growth curve, colony-formation efficiency and the inoculated tumors in nude mice. Results：The transfected cells MAp70 stably expressing antisense HSP70 were obtained, the expression of HSP70 decreased 48％. The cell growth rate was significantly declined as compared to that of the control cell. Colony-formation efficiency and repression rate were 6.08％and 62.14％respectively. The tumor forming time in the inoculated nude mice was 15 days and the average weight of the tumors was 108 mg, significantly lower than that of the control cells. Conclusion：Mediated by retroviral vector, antisense HSP70 can be stably expressed in the MCF7/Adr cells, and the proliferation of MCF7/Adr cells in vitro and the growth of inoculated tumor in nude mice were repressed.