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- W2361083570 abstract "Objective To determine the effects of interfering the expression of focal adhesion kinase (FAK) on sensitivity of colon carcinoma HT29 cell line to 5-fluorouracil (5-FU) by raising the sensitivity of multicellular spheroids. Methods The recombinant plasmid pGenesil-l-FAK, with a short-hairpin RNA (shRNA) targeting the FAK gene, was constructed. RT-PCR and Western blotting were performed to detect the efficiency of interference in both monolayer cell and multicellular spheroids. Moreover, the multicellular spheroids were counted with hemocytometer, and flow cytometry was applied to determine apoptosis. Cell Counting Kit-8 (CCK-8) was used to detect the sensitivity of multicellular spheroids to 5-FU. Results Interference targeting FAK by the recombinant plasmids pGenesil-l-FAK obviously suppressed the FAK mRNA expression in monolayer cells and the protein expression in both monolayer cell and multicellular spheroids. The proliferation velocity of multicelluar spheroids was not interfered by FAK shRNA, and no significant difference on the velocity existed between respective group at different time points (P0.05). Flow cytometry showed that FAK shRNA promoted both the spontaneous apoptosis (16.48%±1.18%) and 5-FU-induced apoptosis (21.50%±2.62%) of multicellular spheroids, and significant difference existed between the two types of apoptosis (P=0.003). Detection for sensitivity by CCK-8 indicated that the sensitivity of both monolayer cells and multicellular spheroids to 5-FU was raised with no significant difference (P0.05) by FAK shRNA interference, thus reversed the resistance induced by the 3D spheroid structure. Conclusion FAK may be involved in the multicelluar resistance of multicelluar spheroids, of which apoptosis and sensitivity to 5-FU are induced by FAK-targeted RNA interference, though the interference might throw no influence on the proliferation." @default.
- W2361083570 created "2016-06-24" @default.
- W2361083570 creator A5058631993 @default.
- W2361083570 date "2009-01-01" @default.
- W2361083570 modified "2023-09-24" @default.
- W2361083570 title "Interference effects of short-hairpin RNA on focal adhesion kinase on the sensitivity to 5-fluorouracil of multicellular spheroids of colonic carcinoma" @default.
- W2361083570 hasPublicationYear "2009" @default.
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