Xu Rong-Hua, Zheng Liang-Yan, He Dong-Lei, Tong Jian, Zheng Li-Ping, Zheng Wu-Ping, Meng Jin, Xia Li-Ping, Wang Cong-Jun, Yi Ji-Lin
epartment of Oncology Surgery, The Affiliated Hospital of Hainan Medical College, Haikou 570102, Hainan Province, China.
World J Gastroenterol. 2008 Jun 21;14(23):3754-8. doi: 10.3748/wjg.14.3754.
To evaluate the inhibitory effects of human fragile histidine triad (FHIT) gene on cell proliferation and apoptosis in human hepatocellular carcinoma line Hep3B in vitro.
A recombinant pcDNA3.1 (+)/FHIT including the functional region of FHIT gene was constructed and transferred into human hepatocellular carcinoma cells in vitro. mRNA and protein expression of the FHIT gene in the transfected cells was detected by RT-PCR and Western blot, respectively. The effect of FHIT on proliferation was detected by MTT assay. Changes in cell cycle and apoptosis were assayed by flow cytometry. Five mice received subcutaneous transplantation of Hep3B-FHIT; 5 mice received subcutaneous transplantation of normal Hep3B and Hep3B-C as controls. The body weight of nude mice and tumor growth were measured.
RT-PCR and Western blot analysis showed that the expression level of FHIT-mRNA and FHIT protein was higher in Hep3B cells after infection with pcDNA3.1 (+)/FHIT. The growth of Hep3B cells treated with pcDNA3.1 (+)/FHIT was significantly inhibited. The pcDNA3.1 (+)/FHIT-transfected Hep3B cells showed a significantly higher cell rate at G(0)-G(1) phase and increased apoptosis in comparison with controls (P < 0.05). The growth of transplanted tumor was inhibited markedly by FHIT. Tumors arising from the Hep3B-FHIT cells occurred much later than those arising from the Hep3B and Hep3B-C cells. The growth of Hep3B-FHIT cells was slow and the tumor volume was low.
Transduction of FHIT gene inhibits the growth of human hepatocellular carcinoma cells and induces cell apoptosis in vivo and in vitro.
评估人脆性组氨酸三联体（FHIT）基因对人肝癌细胞系Hep3B体外增殖和凋亡的抑制作用。
构建含FHIT基因功能区的重组质粒pcDNA3.1(+)/FHIT，并体外转染人肝癌细胞。分别采用RT-PCR和Western blot检测转染细胞中FHIT基因的mRNA和蛋白表达。采用MTT法检测FHIT对细胞增殖的影响。通过流式细胞术检测细胞周期和凋亡的变化。5只小鼠皮下移植Hep3B-FHIT；5只小鼠皮下移植正常Hep3B和Hep3B-C作为对照。测量裸鼠体重和肿瘤生长情况。
RT-PCR和Western blot分析显示，pcDNA3.1(+)/FHIT感染后的Hep3B细胞中FHIT-mRNA和FHIT蛋白表达水平升高。pcDNA3.1(+)/FHIT处理的Hep3B细胞生长明显受到抑制。与对照组相比，pcDNA3.1(+)/FHIT转染的Hep3B细胞在G(0)-G(1)期细胞率显著升高，凋亡增加（P<0.05）。FHIT显著抑制移植瘤的生长。Hep3B-FHIT细胞形成的肿瘤比Hep3B和Hep3B-C细胞形成的肿瘤出现得晚得多。Hep3B-FHIT细胞生长缓慢，肿瘤体积小。
FHIT基因转导在体内外均能抑制人肝癌细胞生长并诱导细胞凋亡。
