自噬基因Beclin1抑制宫颈癌HeLa细胞裸鼠移植瘤生长的研究

Inhibitory effect of autophagy gene Beclin1 on growth of cervical cancer HeLa cells in nude mice.

目的:研究自噬基因Beclin1对裸鼠宫颈癌细胞株HeLa移植瘤的抑制作用。方法:将自噬基因Beclin1真核表达质粒pcDNA3.1(+)-Beclin1经脂质体包裹后转染人宫颈癌HeLa细胞,通过荧光定量PCR(RT-PCR)和Westernblot检测其mRNA及蛋白水平的表达,MTT法检测细胞生长抑制率;将HeLa细胞接种于裸鼠右前肢腋下建立裸鼠人宫颈癌移植瘤模型,待瘤结节直径≥5mm后,分别以重组质粒pcDNA3.1(+)-Bec-lin1、pcDNA3.1(+)和生理盐水200μl每两天1次直接瘤内注射,连用3周后,测量3组肿瘤大小并观察肿瘤组织形态学改变。结果:pcDNA3.1(+)-Beclin1能使HeLa中Bec-lin1mRNA及蛋白表达增加(P<0.05),并显著抑制HeLa细胞的生长,呈现时间依赖性,与其余两组相比,有显著统计学差异(P<0.05);经重组质粒治疗21天后,裸鼠皮下移植瘤生长缓慢,相对于对照组瘤体明显缩小、瘤重明显减轻(P<0.05),病理检查可见,pcD-NA3.1(+)-Beclin1处理组瘤块内细胞坏死减少,结缔组织增生,而对照组中癌细胞分布呈弥漫性。结论:自噬基因Beclin1可以抑制HeLa移植瘤的生长。

Objective：To investigate the inhibitory effects of autophagy gene Beclinl on growth of cervical cancer HeLa cells in nude mice. Methods： The eukaryotic expression vector of Beclinl was transfected via lipofectamine into HeLa cells. Real time-ploymerase chain reaction（RT-PCR） and Western blot were used for detecting expression of mRNA and protein of Beclinl in transfected cells. MTT assay was employed to observe the effect of on the proliferation of HeLa. In vivo study was carried out by injected recombinant Beclinl plasmids into transplanted tumors of nude mice. The tumor-bearing mice were randomized into 3 groups ： pcDNA3. 1 （＋） -Beclinl group, pcDNA3.1 （＋） group and HeLa group, pcDNA3.1 （＋） -Beclinl, pcDNA3. 1 （＋） , and saline were separately injected into the tumor bodies of mice in relevant groups for 3 week. The tumor weight and volume were measured, and the morphology of tumor cells was observed under microscope. Results ： The eukaryotic expression vector pcDNA3. 1 （ ＋ ） -Beclinl significantly improved the expressin of mRNA and protein of Beclinl in HeLa cells（P〈0.05） , the cell proliferations of HeLa cell were inhibited in a time-dependent manner. The tumor growth was significantly inhibited and the tumor weight was higher in pcDNA3.1 （ ＋）-Beclinl group than in the other control groups（ P〈0.05 ）. Less necrosis foci was observed in tumor tissues of pcDNA3.1 （＋）-Beclinl group,while cancer cells were diffused in tumor tissues of pcDNA3.1 （＋） group and HeLa group. Conclusion： Autophagy gene Beclinl can inhibit HeLa cells transplanted tumor growth in nude mice.