腺病毒介导的反义c-myc基因转染抑制人晶状体上皮细胞系HLE-B3的增殖

[Inhibition of proliferation of human lens epithelial cell line HLE-B3 by adenovirus-mediated transfer of antisense c-myc gene].

作者信息

Sun Jie, Xie Li-xin, Wang Yao, Liu Ting

机构信息

Shandong Eye Institute and Hospital, Qingdao 266071, China.

出版信息

Zhonghua Yan Ke Za Zhi. 2005 Feb;41(2):161-5.

PMID:15840345

Abstract

OBJECTIVE

To investigate the effects of adenovirus-mediated transfer of antisense c-myc gene on proliferation and survival of human lens epithelial cell.

METHODS

Immortalized human lens epithelial cell line (HLE-B3) was infected by replication-deficient adenovirus of antisense c-myc gene (Ad-AS-myc). The expression of c-myc mRNA and protein was detected by RT-PCR and Western blotting respectively. Cell proliferation was analyzed by cell growth curve and MTT colorimetric assay, cell death and cell cycle of HLE-B3 were examined by flow cytometry.

RESULTS

More than 80% of HLE-B3 cells were infected successfully after 48 hours incubation with Ad-AS-myc adenovirus. The expression of c-myc mRNA and protein was decreased. Ad-AS-myc suppressed the growth rate, enhanced the cell death (18.33% after 48 hours and 26.93% after 96 hours transduction). The proportions of G1 phase cells were increased to (60.50 +/- 7.59)% after 48 hours and (81.90 +/- 8.60)% after 96 hours transduction, while the proportions of S phase cells were decreased to (28.40 +/- 3.38)% after 48 hours and (16.75 +/- 8.97)% after 96 hours infection.

CONCLUSIONS

Adenovirus-mediated antisense c-myc may inhibit proliferation and induce apoptosis of HLE-B3.

摘要

目的

研究腺病毒介导的反义c-myc基因转染对人晶状体上皮细胞增殖和存活的影响。

方法

用携带反义c-myc基因的复制缺陷型腺病毒（Ad-AS-myc）感染永生化人晶状体上皮细胞系（HLE-B3）。分别采用RT-PCR和蛋白质印迹法检测c-myc mRNA和蛋白的表达。通过细胞生长曲线和MTT比色法分析细胞增殖情况，采用流式细胞术检测HLE-B3细胞的死亡情况和细胞周期。

结果

用Ad-AS-myc腺病毒孵育48小时后，80%以上的HLE-B3细胞被成功感染。c-myc mRNA和蛋白的表达降低。Ad-AS-myc抑制了细胞生长速度，增加了细胞死亡（转导48小时后为18.33%，96小时后为26.93%）。转导48小时后G1期细胞比例增加到（60.50±7.59）%，96小时后增加到（81.90±8.60）%，而S期细胞比例在48小时后降至（28.40±3.38）%，感染96小时后降至（16.75±8.97）%。