[Effect of ST6Gal I siRNA-mediated gene silencing on the adhesion and invasion of SW480 cells].
OBJECTIVE
To study the effect of synthesized ST6Gal I specific siRNA on the adhesion and invasiveness of human colon carcinoma cell line SW480 with over expression of ST6Gal I.


METHODS
A double strand small interference RNA (siRNA) targeting ST6Gal I was designed and synthesized, and then transfected into SW480 cells by lipofectmine 2000. SW480 cells were cultured and divided into 4 groups: blank control group, liposome control group, non-specific siRNA group and ST6Gal I siRNA group. The expression of ST6Gal I mRNA was examined by RT-PCR and the amount of alpha-2, 6-sialylation on the SW480 cell surface was detected by flow cytometry. The adhesion and invasion of SW480 cells to extracellular matrix (ECM) were analyzed by using CytoMatrix kit and cell invasion assay kit, respectively.


RESULTS
After SW480 cells were transfected for 48 hours, the expression of ST6Gal I mRNA in ST6Gal I siRNA group was significantly decreased compared with that in the blank control group, liposome control group, and non-specific siRNA group (P<0.05). After SW480 cells were transfected for 72 hours, the amount of alpha-2, 6-sialylation on cell surface, the adhesion and invasion of the cells in ST6Gal I siRNA group were markedly lower than those in the other 3 groups (P<0.05).


CONCLUSIONS
The chemically synthesized specific siRNA targeting ST6Gal I can effectively inhibit the expression of ST6Gal I and reduce cell adhesion and invasion to ECM in SW480 cells. Our research is important for further study of anti-tumor treatment with RNA interference.
Yuan
Tian-hong
TH
Department of Microbiology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China. gy.th@163.com
Li
Ming-yuan
MY
Li
Wan-yi
WY
Li
Hong
H
Jiang
Zhong-hua
ZH
D002448:Cell Adhesion / Q000235:genetics; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000235:genetics / Q000628:therapy; D005434:Flow Cytometry; D020868:Gene Silencing / Q000502:physiology*; D006801:Humans; D008055:Lipids / Q000235:genetics; D009361:Neoplasm Invasiveness / Q000235:genetics; D034622:RNA Interference / Q000502:physiology; D034741:RNA, Small Interfering / Q000235:genetics / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012799:Sialyltransferases / Q000235:genetics*; D014162:Transfection; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase
D008055:Lipids; C086724:Lipofectamine; D034741:RNA, Small Interfering; D012799:Sialyltransferases; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase