IL-24 基因对 CD133(+)喉癌细胞的抑制作用。

Inhibitive effect of IL-24 gene on CD133(+) laryngeal cancer cells.

机构信息

Second Affiliated Hospital of Jilin University, 130041, Jilin, China.

出版信息

Asian Pac J Trop Med. 2014 Nov;7(11):867-72. doi: 10.1016/S1995-7645(14)60151-6. Epub 2014 Oct 22.

DOI:10.1016/S1995-7645(14)60151-6

PMID:25441985

Abstract

OBJECTIVE

To explore the inhibitive and apoptosis inductive effect of IL-24 genes on CD133(+) laryngeal cancer cells in Hep-2 line.

METHODS

Human peripheral blood monocytes were isolated. The total RNA was extracted by using Trizol method and reverse transcripted into cDNA using RT-PCR method. Primers P1 and P2 was designed for the amplification of human IL-24 genes. After confirmation of agarose gel electrophoresis tests, TA was cloned into pMD19-T simple vector. Nhe I and Xho I double digesting human IL-24 and pIRES2-ZsGreen1 and eukaryotic expression vector were used to establish the pIRES2-ZsGreen1-hIL-24 vector, and detected by enzyme digestion and gene sequencing methods. Flow cytometry (FCM) was used to isolate CD133(+) cells from Hep-2 cells. CD133(+) cells were transfected with pIRES2-ZsGreen1-hIL-24 through liposome 2000. After detection, MTT and FCM were used to observe the effect of IL-24 gene on CD133(+) laryngeal cancer Hep-2 cells.

RESULTS

Lipotin mediated transfection of recombinant pIRES2-ZsGreen1-hIL-24 plasmid into CD133(+) Hep-2 could expressed IL-24 gene in cells stably. MTT results showed that IL-24 transfected group was significantly suppressed compared to empty vector group and control group (P<0.05); FCM results showed that the apoptosis rate of experimental group increased significantly compared to empty vector group and control group (P<0.05).

CONCLUSIONS

IL-24 gene expressions can inhibit proliferation of CD133(+) laryngeal cells in Hep-2 line and promote their apoptosis.

摘要

目的

探讨白细胞介素 24（IL-24）基因对 Hep-2 喉癌细胞系 CD133(+)细胞的抑制和凋亡诱导作用。

方法

分离人外周血单核细胞，采用 Trizol 法提取总 RNA，RT-PCR 法反转录成 cDNA，设计引物 P1 和 P2 扩增人 IL-24 基因，经琼脂糖凝胶电泳验证后，TA 克隆至 pMD19-T 简单载体，Nhe I 和 Xho I 双酶切人 IL-24 与 pIRES2-ZsGreen1 真核表达载体，构建 pIRES2-ZsGreen1-hIL-24 载体，经酶切及基因测序鉴定。流式细胞术（FCM）分离 Hep-2 细胞中的 CD133(+)细胞，脂质体 2000 转染 pIRES2-ZsGreen1-hIL-24 后，通过 MTT 及 FCM 观察 IL-24 基因对 CD133(+)喉癌细胞 Hep-2 的作用。

结果

Lipofectamine 介导重组 pIRES2-ZsGreen1-hIL-24 质粒转染至 CD133(+) Hep-2 细胞后，细胞内可稳定表达 IL-24 基因。MTT 结果显示，与空载体组和对照组比较，IL-24 转染组细胞增殖明显受到抑制（P<0.05）；FCM 结果显示，实验组细胞凋亡率明显高于空载体组和对照组（P<0.05）。