摘要
目的通过构建含有重组AFP(recombinant alpha-fetoprotein,rAFP)启动子的携蜂毒素(melittin,Mel)基因的重组腺病毒(adenovirus,Ad),观察感染Ad-rAFP-Mel的HepG2细胞形态学变化。方法采用倒置显微镜、苏木精-伊红(hematoxylin-eosin,HE)染色、Hoechst 33258染色、透射电镜观察Ad-rAFP-Mel作用过的肝癌细胞死亡情况、细胞器的结构变化等形态学变化。结果倒置显微镜下观察结果显示,随着病毒感染复数的增加和时间的延长,活细胞数量减少;HE染色显示坏死与凋亡并存;Hoechst 33258染色显示确有凋亡存在;透射电镜证实Ad-rAFP-Mel作用于肝癌细胞后,肝癌细胞凋亡和坏死兼而有之。结论Ad-rAFP-Mel能够抑制HepG2细胞的生长,细胞死亡呈现坏死与凋亡兼有的特征。
Objective To observe morphological changes of hepatocarcinoma cell line HepG2 transfected by recombinant adenovirus carrying melittin gene and recombinant alpha-fetoprotein promoter （Ad-rAFP- Mel）. Methods The apoptosis and necrosis as well as structural changes of organelles of Ad-rAFP-Mel transfected HepG2 cells were observed by using inverted microscope, hematoxylin-eosin （HE） staining, Hoechst 33258 staining and transmission electron microscope. Results It was showed under inverted microscope that the number of viable cells declined with the increasing of time and multiplicity of infection. Both apoptosis and nectosis cells were found by HE staining, then the apoptosis of HepG2 cells was proved by Hoechst 33258 staining. Both apoptosis and necrosis of Ad-rAFP-Mel transfected HepG2 cells were proved under transmission electron microscope. Conclusion Ad-rAFP-Mel could inhibit the growth of HepG2 cells, and induce the cell death, including both necrosis and apoptosis.
出处
《安徽中医学院学报》
CAS
2010年第4期45-48,共4页
Journal of Anhui Traditional Chinese Medical College
基金
国家自然科学基金(30600808
30772887)
关键词
肝癌
蜂毒素基因
腺病毒
形态学
凋亡
坏死
Hepatocarcinoma
Melittin gene
Adenovirus
Morphology
Apoptosis
Necrosis
