[Retrovirus-mediated IL-18 gene expression in rat C6 glioma cell line].
OBJECTIVE
To set up rat C6 glioma cell line C6/IL-18 expressing IL-18 gene and explore the effect of IL-18 on the growth of C6 cells.


METHODS
The IL-18 gene was transferred into the C6 cells by a retrovirus vector. After screening with G418, the C6/IL-18 cells were obtained. IL-18 mRNA expression in C6/IL-18 cells was detected with RT-PCR. The expression of IL-18 protein was detected by flow cytometry and immunocytochemical staining. In order to analyze the activity of the expressed IL-18 protein, ELISA was used to detect the ability to secrete IFN-gamma by rat splenocytes induced with the culture supernatant of C6/IL-18 cells. The in-vitro proliferation of C6/IL-18 cells was detected by MTT colorimetry and flow cytometry. The rat model was used to observe the tumorigenic activity of the C6/IL-18 cells.


RESULTS
IL-18 mRNA and protein were stably expressed in C6/IL-18 cells. The culture supernatant of C6/IL-18 cells induced secretion of IFN-gamma by rat splenocytes. At the same time, the proliferation rate and in-vivo tumorigenicity of C6/IL-18 cells were markedly reduced as compared with parental C6 cells.


CONCLUSIONS
Exogenous IL-18 gene can inhibit the proliferation and in-vivo tumorigenicity of C6 cells.
Li
Wen-ling
WL
Cell Biology Division, Institute of Basic Medicine, Hebei Medical University, Shijiazhuang 050017, China.
Yan
Yun-li
YL
Shan
Bao-an
BA
Zheng
Li-fen
LF
Li
Qiao-xia
QX
Zhou
Na-jing
NJ
D000818:Animals; D001932:Brain Neoplasms / Q000378:metabolism / Q000473:pathology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D017077:Culture Media, Conditioned / Q000494:pharmacology; D005822:Genetic Vectors; D005910:Glioma / Q000378:metabolism / Q000473:pathology*; D007371:Interferon-gamma / Q000378:metabolism; D020382:Interleukin-18 / Q000096:biosynthesis* / Q000235:genetics; D008297:Male; D009368:Neoplasm Transplantation; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D012190:Retroviridae / Q000235:genetics*; D013154:Spleen / Q000166:cytology / Q000378:metabolism
D017077:Culture Media, Conditioned; D020382:Interleukin-18; D012333:RNA, Messenger; D011994:Recombinant Proteins; D007371:Interferon-gamma