摘要
目的探讨携带人基质金属蛋白酶组织抑制因子-1(hTIMP-1)的重组腺病毒(AdhTIMP-1)对人肝癌HepG2细胞增殖、侵袭、转移、凋亡以及瘤内血管生成的作用。方法构建AdhTIMP-1并感染人肝癌HepG2细胞,采用Boyden chamber检测细胞体外侵袭能力;感染AdhTIMP-1的HepG2细胞接种于裸鼠皮下观察其成瘤情况;建立荷瘤鼠模型,瘤内注射AdhTIMP-1,观察其对肿瘤生长的作用;处死荷瘤鼠,计数肺转移结节;CD34免疫组化观察肿瘤血管生成;原位末端标记(TUNEL)法检测肝癌细胞凋亡。结果AdhTIMP-1感染的HepG2细胞体外侵袭力下降91.4%,成瘤量下降75.8%,荷瘤鼠瘤体内注射AdhTIMP-1使肿瘤生长减少45.4%,组织血管密度减少47.8%,肺转移结节减少70.4%,肿瘤组织中细胞凋亡则增加了3倍。结论AdhTIMP-1能抑制肝癌HepG2细胞的侵袭、转移及瘤内血管形成,诱导肝癌细胞凋亡,可用于肝癌基因治疗的研究。
Objective To explore the effects of overexpression of human tissue inhibitors of metalloproteinase-1 （hTIMP-1） on proliferation, invasion, metastasis, angiogenesis, and apoptosis in human hepatocellular carcinoma （HCC） cells in vitro and in vivo. Methods Recombinant adenoviral vector containing hTIMP-1 （ AdhTIMP-1 ） was constructed previously. HepG2 cells were infected by AdhTIMP-1 and the changes of cell proliferation and invasion were detected in vitro. The anticaneer activity of AdhTIMP-1 was evaluated in BAL B/c mice bearing HCC. Tumor volume and pulmonary metastases were observed. The mechanisms underlying the antitumor effect in vivo were investigated based on detection of microvessel density and apoptosis in tumor tissues. Results The resultant AdhTIMP-1 was successfully constructed and the expression of hTIMP-1 was detected by Western blot and RT-PCR. AdTIMP-1 could effectively infect HepG2 cells and significantly inhibit the proliferative activity and invasive ability of the tumor ceils. Compared with the controls, pre-infection of HepG2 cells by AdhTIMP-1 resulted in a significant inhibition of tumor formation by 75.8%. A single local injection of AdhTIMP-1 into pro-established tumors significantly reduced the tumor growth rate by 45.4%, tumor-associated angiogenesis index by 47.8%, lung metastases by 70.4%, and showed a 3-fold increase of apoptotic tumor cells. Condusion Our data indicated that AdhTIMP-I can significantly attenuate tumor proliferation and invasion, reduce metastasis, inhibit angiogenesis, and induce apoptosis in HCC-bearing mice and may pave the way for further liver cancer gene therapy.
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2007年第11期813-817,共5页
Chinese Journal of Oncology
基金
四川省教育厅基金资助项目(2006B108)
