白细胞介素—6反义基因对人肺癌细胞生长的抑制作用

目的通过体外实验证实白细胞介素－６（ｉｎｔｅｒｌｅｕｋｉｎ－６,ＩＬ－６）为人肺癌细胞系ＰＧ自分泌生长刺激因子,本研究旨在进一步探讨ＩＬ－６对ＰＧ细胞体内生长的影响。方法构建ＩＬ－６反义基因表达载体,应用磷酸钙沉淀法导入ＰＧ细胞以降低内源性ＩＬ－６表达;应用ＲＮＡ斑点杂交检测ＩＬ－６反义ｍＲＮＡ表达;应用生物活性检测法测定ＩＬ－６分泌,裸鼠皮下接种检测细胞成瘤性。结果转染反义ＩＬ－６基因的细胞ＰＧＴＡＳ１、ＰＧＴＡＳ６、ＰＧＴＡＳ８、ＰＧＴＡＳ９能够表达较高水平ＩＬ－６反义ｍＲＮＡ,所分泌的有生物活性的ＩＬ－６降低;ＰＧＴＡＳ１、ＰＧＴＡＳ６、ＰＧＴＡＳ８、ＰＧＴＡＳ９细胞体外生长减慢;ＰＧ－ＴＡＳ６、ＰＧＴＡＳ８、ＰＧＴＡＳ９细胞体内成瘤延迟,生长减慢。反义ＩＬ－６对ＰＧ细胞生长的抑制作用与ＩＬ－６分泌量呈负相关。结论ＩＬ－６对ＰＧ细胞生长具有重要的调控作用,ＩＬ－６为ＰＧ细胞自分泌生长刺激因子。

The inhibitory effect of antisense interleukin-6 on the growth of human lung carcinoma cells in vivo

Objective Interleukin 6(IL 6) has previously been implicated as a potential positive autocrine regulator of a human lung carcinoma cell lince PG. The purpose of the present study is to further evaluate the role of IL 6 in PG cells in vivo.Methods An antisense expression vector for IL 6 was constructed and introduced into PG cells. Antisense mRNA of IL 6 was detected by RNA RNA dot blotting analysis. The production of bioactive IL 6 was measured by bioassay method. To determine the effect of antisense IL 6 cDNA on tumorigenicity, PG cells were inoculated subcutaneously into nude mice.Results Four transfectants, PGTAS1, PGTAS6, PGTAS8, PGTAS9, could express IL 6 antisense mRNA and secret decreased bioactive IL 6. The growth rate of 4 transfectants revealed was reduced in vitro. Comparing with the mice injected with the control PGTneo cells, the latent period of the mice inoculated with PGTAS6,PGTAS8, PGTAS9 cells was significantly increased, and the growth rate of the tumors was obviously decreased. The tumor size was markedly smaller. There was a negative correlation between the growth inhibiting effect of IL 6 antisense gene and the amount of IL 6 secretion. Conclusion These results indicate that IL 6 can function as an autocrine stimulator for PG cells in vivo as well as in vitro.