[ST6Gal I siRNA and antisense oligonucleotide-mediated gene silencing lowers the invasiveness potential of colonic carcinoma cells].

Abstract

OBJECTIVE:

To study the effects of small interfering RNA (siRNA) and antisense oligonucleotides (ASOs) targeting ST6Gal I on adhesion and invasiveness of human colonic carcinoma cell line SW480 over-expressing ST6Gal I.

METHODS:

siRNA and ASOs targeting ST6Gal I were constructed and transfected into SW480 cells via lipofectmine 2000. SW480 cells were cultured and divided into 7 groups, namely the blank control group, liposome group, siRNA group (transfected with ST6Gal I siRNA), ASO(1) group (transfected with ST6Gal I ASO whose target site is different from the siRNA), ASO(2) group (transfected with ST6Gal I ASO targeting the same site as siRNA), siRNA+ASO(1) group (transfected with siRNA and ASO(1)), siRNA+ASO(2) group (transfected with siRNA and ASO(2)). RT-PCR was used to examine ST6Gal I mRNA expression following the treatment. Flow cytometry was used to examine the amount of alpha2,6-sialylation on SW480 cell surface. SW480 cell adhesion and invasiveness to the extracellular matrix (ECM) were analyzed using CytoMatrix kit and cell invasion assay kit, respectively.

RESULTS:

The expression of ST6Gal I mRNA, the amount of alpha2,6-sialylation on the cell surface and cell adhesion and invasion to ECM decreased remarkably in groups siRNA, ASO(1), ASO(2), siRNA+ASO(1) and siRNA+ASO(2), all significantly lower than those of the blank control and liposome groups (all P<0.05), especially in siRNA+ASO(1) group. Significant difference was noted between siRNA+ASO(1) and siRNA groups (P<0.05), but not between siRNA+ASO(2) and siRNA groups, or between blank control and liposome groups (all P>0.05).

CONCLUSION:

Chemically synthesized specific siRNA targeting ST6Gal I effectively inhibits SW480 cell ST6Gal I expression and leads to diminished cell adhesion and invasiveness to ECM, suggesting a combined effect of siRNA and ASO with different targeting sites.

• Authors: Tian-Hong Yuan, Ming-Yuan Li, Wan-Yi Li, Hong Li, Zhong-Hua Jiang
• Journal: Nan fang yi ke da xue xue bao = Journal of Southern Medical University (Nan Fang Yi Ke Da Xue Xue Bao) Vol. 27 Issue 2 Pg. 136-40 (Feb 2007) ISSN: 1673-4254 [Print] China
• PMID: 17355919 (Publication Type: English Abstract, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Oligonucleotides, Antisense
• RNA, Messenger
• RNA, Small Interfering
• Sialyltransferases
• beta-D-Galactoside alpha 2-6-Sialyltransferase

Topics

• Cell Adhesion
• Cell Movement
• Colonic Neoplasms (genetics, pathology)
• Flow Cytometry
• Gene Silencing
• Humans
• Neoplasm Invasiveness
• Oligonucleotides, Antisense (genetics)
• RNA, Messenger (genetics, metabolism)
• RNA, Small Interfering (genetics)
• Reverse Transcriptase Polymerase Chain Reaction
• Sialyltransferases (genetics, metabolism)
• Transfection
• beta-D-Galactoside alpha 2-6-Sialyltransferase