[Short hairpin RNA-mediated survivin gene silencing inhibits invasion and metastasis of human colon carcinoma cell line SW480 in vitro].
OBJECTIVE
To investigate the effect of short hairpin RNA (shRNA) targeting survivin on adhesion and invasion of human colon carcinoma cell line SW480 in vitro.


METHODS
According to the sequence of the coding region of survivin gene, two strings of 19 nucleotides of inverted sequence flanking the loop sequence of two complementary 9-base oligonucleotides were designed and synthesized to prepare the hairpin construct as the DNA templates for the target shRNA. The shRNA templates were cloned into shRNA expression vector pRNAT-U6.1/Neo, and the resulted vector pRNAT-U6.1/Neo-survivin was transfected into SW480 cells using Lipofectamine 2000. Western blotting was performed to evaluate survivin gene silencing induced by shRNA transfection at the protein level, and the biological behaviors of the SW480 cells were investigated by cell-matrix adhesion, invasion and gelatin-zymography assays.


RESULTS
Western blotting revealed significantly lowered survivin protein expression in transfected SW480 cells, and survivin gene silencing induced by shRNA significantly suppressed the metastatic potential of SW480 cells in association with suppressed MMPs activity.


CONCLUSIONS
Survivin may play an important role in modulating human colorectal carcinoma cell invasion and metastasis, and survivin gene silencing can inhibit human colorectal cancer cell invasion and the production of MMP-2 and MMP-9. Survivin may affect invasion and metastasis of human colorectal carcinoma cells via regulating the production of MMPs.
Institute of Digestive Diseases, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. jqb@fimmu.com
Jin-bao
JB
Wu
Qing-zhen
QZ
Nan
Gao-feng
GF
Ma
Wei
W
Gong
Lin
L
Chen
Ying-zhuo
YZ
Lin
Ji-de
JD
Wang
Hong-quan
HQ
Zhang
Yu-gang
YG
Song
D000818:Animals; D015153:Blotting, Western; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000235:genetics / Q000473:pathology*; D020868:Gene Silencing; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins; D055029:Inverted Repeat Sequences; D020782:Matrix Metalloproteinases / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000172:deficiency* / Q000235:genetics*; D009361:Neoplasm Invasiveness / Q000235:genetics; D009362:Neoplasm Metastasis / Q000235:genetics*; D034741:RNA, Small Interfering / Q000235:genetics*; D000077022:Survivin
C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins; D034741:RNA, Small Interfering; D000077022:Survivin; D020782:Matrix Metalloproteinases