Effects of ST6Gal I antisense oligonucleotide-mediated gene silencing on cell adhesion and invasiveness of hela cells.
- Author:
Tian-hong YUAN
1
;
Ming-yuan LI
;
Wan-yi LI
;
Hong LI
;
Zhong-hua JIANG
Author Information
- Publication Type:Journal Article
- MeSH: Cell Adhesion; genetics; physiology; Cell Movement; genetics; physiology; Flow Cytometry; Gene Silencing; HeLa Cells; Humans; Neoplasm Invasiveness; Oligodeoxyribonucleotides, Antisense; genetics; RNA, Messenger; biosynthesis; genetics; Reverse Transcriptase Polymerase Chain Reaction; Sialyltransferases; biosynthesis; genetics; Transfection
- From: Chinese Journal of Oncology 2007;29(1):21-24
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo study the effects of antisense oligonucleotide (ASODN) targeting ST6Gal I on cell adhesion and invasiveness of human cervical carcinoma cell line HeLa which over-expressed ST6Gal I .
METHODSASODN and sense oligonucleotide (SODN) targeting ST6Gal I were designed and constructed, and transfected into a cervical cancer cell line, HeLa, by lipofectmine 2000. HeLa cells were cultured and divided into 4 groups: blank control group, liposome group, SODN group and ASODN group. RT-PCR was used to examine the ST6Gal I mRNA expression. Flow cytometry was used to examine the amount of alpha2, 6-sialylation on the HeLa cell surface. The HeLa cell adhesion and invasiveness to extracellular matrix ( ECM) were analyzed by using CytoMatrixTM kit and cell invasion assay kit, respectively.
RESULTSThe expression of ST6Gal I mRNA in HeLa cells at 48 hrs after transfection in the ASODN group was significantly decreased in comparison with that in the blank control group, liposome group, and SODN group(P <0. 01). The amount of alpha2,6-sialylation on cell surface in ASODN group was significantly lower than that of the other 3 groups ( P <0. 05). The adhesion and invasiveness of the cells in the ASODN group decreased remarkably, both significantly lower than those of the other 3 groups ( all P < 0. 05).
CONCLUSIONSpecific ASODN targeting ST6Gal I effectively inhibits HeLa cell ST6Gal I expression, decreases the amount of alpha2,6-sialylation on cell surface and leads to a decline of cell adhesion and invasiveness to ECM. This result also established a fine base for further studying on anti-tumor treatment with antisense oligonucleotide.