[Construction of adenovirus expressing siRNA against hTR and its anti-tumor activity in vitro].
OBJECTIVE
To study the effect of the hTR-siRNA adenovirus on hTR mRNA gene silence, telomerase activity inhibition and anti-tumor in vitro.


METHODS
RNAi adenovirus vector, Ad-hTR-siRNA, and negative control Ad-NT-siRNA were constructed by an improved ligation method. Different tumor cells and liver cell line, HL-7702, were infected with 100 MOI of the recombinant adenoviruses. TRAP-ELISA, Real-time PCR and FCM were used to analyze telomerase activity, hTR mRNA, apoptosis rate and hTERT protein expression.


RESULTS
As compared with Ad-NT-siRNA, Ad-hTR-siRNA reduced both hTR mRNA levels (70.21%) and telomerase activity (58.87%) of HeLa cells significantly, increased apoptosis rate (29.7%). But the telomerase activity of HL-7702 and hTERT protein didn't show the tendency of decrease.


CONCLUSIONS
It is supposed that the hTR-siRNA adenovirus could knockdown hTR gene specifically and suppress the tumor cell growth in vitro efficiently. Maybe this siRNA expressing recombinant adenovirus system could be a new method for cancer gene therapy.
Sichuan University, Chengdu, China. feifeiliyan@163.com
Yan
Y
Li
Li-ying
LY
Xiao
Gang
G
Yao
Hong
H
Li
Zhi-wei
ZW
Yang
Wan-yi
WY
Li
Zhong-hua
ZH
Jiang
Ming-yuan
MY
Li
D000256:Adenoviridae / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D049109:Cell Proliferation; D018390:Gene Targeting / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D009369:Neoplasms / Q000201:enzymology* / Q000235:genetics / Q000503:physiopathology; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism; D019098:Telomerase / Q000235:genetics* / Q000378:metabolism
D034741:RNA, Small Interfering; D019098:Telomerase