[Effects of galectin-3 on proliferation and apoptosis of hepatic stellate cells].
OBJECTIVE
To observe the effects of galectin-3 on proliferation and apoptosis of hepatic stellate cells.


METHODS
RT-PCR and Western blot were used to detect the expression of galectin-3 in hepatic stellate cells. Short hairpin DNA targeting galectin-3 of rat was was ligated into the recombinant vector pGCsilencer U6/Neo/GFP/shRNA plasmid. Then the plasmid was transfected into rat hepatic stellate cells. RT-PCR and Western blot were used to detect the interfering efficiency. Cell proliferation level was observed by CCK8 method at 24, 48 and 72 hours after transfection. Cell apoptosis was measured by Annexin V/PI-labeled flow cytometric analysis.


RESULTS
Expression of galectin-3 in HSC was verified by both RT-PCR and Western blot. The recombinant vector was successfully constructed and verified, and was transfected into rat hepatic stellate cells. Western Blot and RT-PCR results demonstrated that the expression level of Galectin-3 was significantly down-regulated in galectin-3 shRNA transfected cells compared to control vector transferred cells. CCK8 assay indicated that proliferation of Galectin-3 knockdown cells was lower than that of control cells 48 and 72 hours post-transfection. Apoptotic cells in shRNA-interfering group were higher than those in control group both in early stage and advanced stage.


CONCLUSIONS
Hepatic stellate cells can express galectin-3. Inhibition of galectin-3 using RNAi technique can suppress proliferation and induce apoptosis in HSC.
Department of Hepatobiliary Surgery, Peking University People's Hospital, Beijing 100044, China.
Yi-fei
YF
Feng
Tao
T
Li
Shu
S
Li
Ji-run
JR
Peng
Xi-sheng
XS
Leng
D000818:Animals; D017209:Apoptosis; D002460:Cell Line; D049109:Cell Proliferation; D015536:Down-Regulation; D005434:Flow Cytometry; D037502:Galectin 3 / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors; D055166:Hepatic Stellate Cells / Q000166:cytology / Q000378:metabolism*; D008103:Liver Cirrhosis / Q000473:pathology; D010957:Plasmids / Q000235:genetics; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics*; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D037502:Galectin 3; D012333:RNA, Messenger; D034741:RNA, Small Interfering