The Effect Of RNA Interference On The Adhesive Ability Of Multi-drug Resistant Ovarian Cancer Cell Line SKOV3-TR30

OBJECTIVES:To research the influence of small interfering RNA on adhesive ability of paclitaxel-resistance human ovarian cancer cell line SKOV3-TR30 in vitro.METHODS:1.siRNAs targeting on the MDR-1 gene sequences were chemically synthesized.Mdr1-siRNAs were transfected into SKOV3-TR30 by lipofectAMINE2000.2.The technique of EDTA-induced cell detachment was used to assay the cell's adhesive ability to plastic substance.3.The cell's adhesive ability to LN and FN-components of basement membrance was measured by MTT assay.4.The expression of CD44v6 protein was detected by Western Blot.5.Immunol Fluorescence Staining was used to view the expression of CD44v6 of these cells.6.The percentage of CD44v6-positive cells was detected by flow cytometry.RESULTS:1.EDTA-induced cell detachment test showed that the MDR-1 siRNA modified SKOV3-TR30 cells adhered less effectively to plastic substance(P＜0.05).2.Test of Cell's adhesion to extracellular matrix and basement membrance showed that SKOV3-TR30/MDR cells' adhesion to LN and FN was significantly inhibited(P＜0.05).3.The expression of CD44v6 protein in the cells detected by Western Blot and Immunol Fluorence Staining were varied significantly. Compared with SKOV3 cells,no significant change was showed in the expression of CD44v6 detected in SKOV3-TR30/MDR cells, and that of SKOV3-TR30 and SKOV3-TR30/non cells were significantly elevated.The percentage of CD44v6-positive cells were 30.72%,31.63%,66.09%and 68.15%respectively,which decreased obviously in SKOV3-TR30/MDR cells.CONCLUSIONS:Mdr1-siRNAs transfected by liposome can reduce the adhesive capacity of paclitaxel-resistance ovarian carcinoma SKOV3-TR30 cells.