Simultaneous silencing of VEGF and KSP by siRNA cocktail inhibits proliferation and induces apoptosis of hepatocellular carcinoma Hep3B cells.
BACKGROUND
Vascular endothelial growth factor (VEGF) is involved in the growth of new blood vessels that feed tumors and kinesin spindle protein (KSP) plays a critical role in mitosis involving in cell proliferation. Simultaneous silencing of VEGF and KSP, an attractive and viable approach in cancer, leads on restricting cancer progression. The purpose of this study is to examine the therapeutic potential of dual gene targeted siRNA cocktail on human hepatocellular carcinoma Hep3B cells.


RESULTS
The predesigned siRNAs could inhibit VEGF and KSP at mRNA level. siRNA cocktail showed a further downregulation on KSP mRNA and protein levels compared to KSP-siRNA or VEGF-siRNA, but not on VEGF expression. It also exhibited greater suppression on cell proliferation as well as cell migration or invasion capabilities and induction of apoptosis in Hep3B cells than single siRNA simultaneously. This could be explained by the significant downregulation of Cyclin D1, Bcl-2 and Survivin. However, no sigificant difference in the mRNA and protein levels of ANG2, involving inhibition of angiogenesis was found in HUVECs cultured with supernatant of Hep3B cells treated with siRNA cocktail, compared to that of VEGF-siRNA.


CONCLUSIONS
Silencing of VEGF and KSP plays a key role in inhibiting cell proliferation, migration, invasion and inducing apoptosis of Hep3B cells. Simultaneous silencing of VEGF and KSP using siRNA cocktail yields promising results for eradicating hepatocellular carcinoma cells, a new direction for liver cancer treatment.
Faculty of Biology, University of Science, Vietnam National University, Ward 4,District 5, 227 Nguyen Van Cu Street, Ho Chi Minh City, Vietnam. dcchung@hcmus.edu.vn.|Department of Animal Biotechnology, Institute of Tropical Biology, Vietnam Academy of Science and Technology, 9/621 Xa lo Ha Noi Street,Linh Trung Ward, Thu Duc District, Ho Chi Minh City, Vietnam. dcchung@hcmus.edu.vn.
Chung Chinh
CC
Doan
Department of Animal Biotechnology, Institute of Tropical Biology, Vietnam Academy of Science and Technology, 9/621 Xa lo Ha Noi Street,Linh Trung Ward, Thu Duc District, Ho Chi Minh City, Vietnam. lelong2510@gmail.com.
Long Thanh
LT
Le
Department of Animal Biotechnology, Institute of Tropical Biology, Vietnam Academy of Science and Technology, 9/621 Xa lo Ha Noi Street,Linh Trung Ward, Thu Duc District, Ho Chi Minh City, Vietnam. hoangnghiason@itb.ac.vn.
Son Nghia
SN
Hoang
Faculty of Biology, University of Science, Vietnam National University, Ward 4,District 5, 227 Nguyen Van Cu Street, Ho Chi Minh City, Vietnam. dmsy@hcmus.edu.vn.
Si Minh
SM
Do
Department of Immunology, Vietnam Military Medical University, 160 Phung Hung Street,Ha Dong District, Ha Noi City, Vietnam. levandong@vmmu.edu.vn.
Dong Van
DV
Le
D017304:Annexin A5; D017209:Apoptosis / Q000235:genetics*; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation / Q000235:genetics*; D019938:Cyclin D1 / Q000378:metabolism; D015853:Cysteine Proteinase Inhibitors / Q000378:metabolism; D015536:Down-Regulation; D005434:Flow Cytometry; D020868:Gene Silencing; D019254:Genes, bcl-2; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000378:metabolism; D016547:Kinesins / Q000235:genetics* / Q000378:metabolism; D008938:Mitosis / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D000077022:Survivin; D013778:Tetrazolium Salts; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics* / Q000378:metabolism; D033921:Vesicular Transport Proteins / Q000378:metabolism
C110656:2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium; D017304:Annexin A5; C107489:BIRC5 protein, human; D015853:Cysteine Proteinase Inhibitors; D051034:Inhibitor of Apoptosis Proteins; C470243:KIF11 protein, human; D034741:RNA, Small Interfering; D000077022:Survivin; D013778:Tetrazolium Salts; C555023:VPS51 protein, human; D042461:Vascular Endothelial Growth Factor A; D033921:Vesicular Transport Proteins; D019938:Cyclin D1; D016547:Kinesins