白细胞介素2基因工程人大肠癌细胞瘤苗的建立及鉴定

Establishment of the interleukin-2 gene transferred human colorectal cancer cell vaccine

目的 建立高表达白细胞介素2的基因工程人大肠癌细胞瘤苗，并完成相关的实验室鉴定。方法 采用重组逆转录病毒将人白细胞介素2基因转导至三株人大肠癌细胞，G418筛选并挑选单克隆，ELISA检测各单克隆24h白细胞介素2表达量，用Student Newman Keuls检验三株细胞白细胞介素2的表达差异，PCR、Southern blot检测白细胞介素2基因整合，RT-PCR、Northern blot检测白细胞介素2基因转录水平。结果 获得72株G418抗性单克隆，其中表达量最高者为COL0205达138.01ng/L×10^7/24h，统计显示COLO205与SW1116、HT-29组间相比差异有统计学意义（P〈0.05），白细胞介素2基因整合至细胞基因组中，未发生重组或部分缺失，且获得成功表达。结论 本研究成功建立了高表达白细胞介素2的基因工程人大肠癌细胞瘤苗。

Objective To establish an interleukin-2 gene transferred human eoloreetal cancer cell vaccine. Methods Interleukin-2 gene was transferred into three human eoloreetal cancer cell lines with reeombined retrovirus. After using G418 to select monoelones, the secretion of interleukin-2 was determined by ELISA. Student Newman Keuls was used to examine the difference of three kinds of eoloreetal cancer cells; PCR,Southem blot to confirm the combination of interleukin-2 gene; RT-PCR,Northem blot to cheek the expression of interleukin-2 gene in mRNA level. Results We obtained 72 monoelones which may contain the objective gene. The highest secretion level was COLO205 of 138.01 ng/L × 10^7/24 hr. There was a significant difference between the production of IL-2 between COLO205 and SW1116, HT-29. We came to the conclusion that interleukin-2 gene was successfully combined to cell genome without recombination and exealation, and was translated correctly. Conclusions we succeeded in establishing the interleukin-2 gene transferred human eoloreetal cancer cell vaccine.