摘要
目的探讨半乳糖受体介导的c-myc反义寡核苷酸(antisenseoligodeoxynucleotide,ASODN)抑制肝癌Bel-7402细胞增殖的机制。方法c-mycASODN与半乳糖(galactose,Gal)-聚乙烯亚胺(poly-ethyleneimine,PEI)相作用形成Gal-PEI-ASODN复合物,作用于人肝癌Bel-7402细胞,通过流式细胞仪检测细胞周期、AnnexinV-FITC和碘化丙啶(PropidiumIodide,PI)双染色、DNA电泳实验观察Gal-PEI-ASODN对Bel-7402细胞的作用。结果采用流式细胞仪检测细胞周期,细胞对照组、Gal-PEI对照组、ASODN对照组,细胞增殖指数分别为35.04%、33.95%、32.90%,Gal-PEI-ASODN组细胞增殖指数为23.65%,与细胞对照组相比,sub-G1期+G0/G1期细胞总数从64.03%增加到76.74%,S期+G2/M期的细胞从35.04%减少为23.65%,Gal-PEI-ASODN组细胞增殖指数下降11.39%,差异显著(P<0.01)。采用细胞凋亡检测试剂AnnexinV-FITC/PI双染检测细胞坏死情况,细胞对照组细胞凋亡率2.77%,坏死率3.06%,正常细胞率94.13%;ASODN对照组、Gal-PEI对照组细胞凋亡率均在4%以下,坏死率均在6.8%以下;Gal-PEI-ASODN组细胞凋亡率6.5%,坏死率15.9%,正常细胞率下降为76%。DNA电泳实验中,Gal-PEI-ASODN组未发现细胞凋亡的DNA梯形条带,相反出现了弥散条带。结论半乳糖受体介导的c-myc反义核酸通过阻止细胞从G0/G1期细胞向S期的转变,诱导细胞坏死,达到抑制细胞Bel-7402增殖的作用。
Objective To observe the mechanism of c-myc antisense oligodeoxynucleotide mediated by galactose receptor on the proliferative inhibition of hepatocarcinoma Bel-7402 cells. Methods c-myc ASODN mixed with Galactose （GAD- polyethyleneimine （PEI） reagent forming Gal-PEI ASODN complex, then incubated with Bel-7402 cells for 48h, using Propidium Iodide（PI） dying, AnnexinV-FITC and PI double staining,DNA gel electrophoresis to detected the proliferative inhibition mechanism of Gal-PEI-ASODN on Bel-7402 cells. Results Cell cycle was determined by flow cytometry. In the control group, Gal-PEI group and ASODN group, cell proliferative index was found to be 35.04%,33. 95%,32. 90%, respectively. However,cell proliferative index for Gal PEI-ASODN group was found to be 23.65%, decreased by 11. 39% compared to the control group（P〈0.01 ）. Necrotic cell percentage was determined using AnnexinV-FITC/ PI double staining test. The results showed cell apoptotic rate of 2. 77%, cell necrotic rate of 3. 06% and the normal cell percentage was detected as 94. 13% in the control group;cell apoptotic rate below 4%, cell necrotic rate below 6.8% were detected in both ASODN group and Gal-PEI group; cell apoptotic rate of 6.5%, cell necrotic rate of 15.9% and the normal cell percentage of 76% were determined in Gal-PEI-ASODN group. There was no DNA ladder morphology, on the contrary, there was DNA smear morphology using DNA gel electrophoresis in Gal-PEI-ASODN group. Conclusion The mechanism by which Gal-PEI-ASODN inhibits cell proliferation maybe that it can restrain the change from G0/G1 period to S period in cell cycle and induces cell necrosis.
出处
《肿瘤防治研究》
CAS
CSCD
北大核心
2006年第10期729-732,共4页
Cancer Research on Prevention and Treatment
基金
广东省自然科学基金重点资助项目(021195)
广东省自然科学基金资助项目(04010487)
关键词
c—myc
反义核酸
肝癌
细胞
坏死
细胞周期
c-myc
Antisense oligodeoxynucleotide
Hepatocellular carcinoma
Necrosis
Apoptosis
作者简介
蒋建伟（1966-），男，博士，副教授，主要从事肿瘤分子生物学的研究
通讯作者
