摘要
目的:研究细胞分裂周期素25a(cell division cycle 25a,CDC25a)基因RNAi(RNA干扰)的重组慢病毒载体对人肝癌细胞Hep G2 CDC25a基因表达和裸鼠移植瘤生长的影响。方法:实验组以CDC25a基因重组慢病毒颗粒感染Hep G2细胞;阴性对照组以RNAi-NC慢病毒颗粒感染Hep G2细胞;空白对照组常规培养。三组细胞分别接种至裸鼠皮下,建立人肝癌裸鼠移植瘤模型。连续28 d观察裸鼠成瘤情况及移植瘤生长情况,28 d后测定肿瘤体积和重量。用RT-PCR、Western blot及免疫组织化学法检测移植瘤中CDC25a的表达情况。结果 :各组裸鼠接种Hep G2细胞后第7天均有肿瘤形成,实验组的肿瘤生长速度、体积和重量明显低于空白对照组和阴性对照组(P<0.05)。实验组瘤体中CDC25a的m RNA及蛋白表达与阴性对照组及空白对照组相比明显下降(P<0.05)。结论:LV-CDC25a-RNAi重组体沉默Hep G2细胞的CDC25a基因后能有效抑制人肝癌裸鼠移植瘤的生长,提示CDC25a基因可被认为是肝癌治疗的关键靶点。
Objective To research the effects of the recombinant lentiviral vector for RNA interference （RNAi）of CDC25a gene on the expression of CDC25a gene and the growth of subcutaneous tumor of human liver cancer HepG2 cells in nude mice. Methods HepG2 cells infected with recombinant CDC25a gene Lentiviral Particles （LV-CDC25A-RNAi） were used to be the experimental group; HepG2 cells infected with RNAi-NC Lentiviral Particles were used to be the negative control group ; the normal control with no treatment. Nude mice were inoculated subcutaneously with the three groups ceils to establish the subcutaneous tumor model of hepatocellular carcinoma. Observed the tumors formation and growth in nude mice for 28 days. The tumors volume and weight were measured after 28 days. To detect the expression of CDC25a in the subcutaneous tumors by transcription polymerase chain reaction （RT-PCR）, Western blot and Immune histochemical method. Results At least 7 days after inoculation of cells were tumor formation. Tumors growth rate, volume and weight in the experimental group were significantly lower than the control and negative control group （P 〈 0.05）. CDC25a mRNA and protein expression in experimental group＇s tumor tissue was lower than the negative control and blank control group （P 〈 0.05）. Conclusion The expression of CDC25a gene could significantly be silenced by using technology of RNAi, the growth of subcutaneous tumor could be inhibited also. It suggests that CDC25a gene can be considered as a key target for the treatment of liver cancer.
出处
《实用医学杂志》
CAS
北大核心
2015年第3期348-352,共5页
The Journal of Practical Medicine
基金
国家自然科学基金项目(编号:30960428)
广西科技基础条件平台建设项目(编号:10-108-25)
广西科学基金项目(编号:2013GXNSFAA019210)
