Effect of RNA interference inhibition to expression of CD133 on tumor cell biological characteristics in KATO-III CD133(+) cells of human gastric cancer

• Published in: Zhonghua wei chang wai ke za zhi Vol. 16; no. 9; p. 889
• Main Authors: Wang, Shou-lian, Yu, Ji-wei, Cai, Cheng, Lu, Rui-qi, Wu, Ju-gang, Ni, Xiao-chun, Jiang, Bo-jian
• Format: Journal Article
• Language: Chinese
• Published: China 01.09.2013
• Subjects: AC133 Antigen; Antigens, CD - genetics; Antigens, CD - metabolism; Cell Line, Tumor; Cell Movement; Cell Proliferation; Fluorouracil - pharmacology; Glycoproteins - genetics; Glycoproteins - metabolism; Humans; Peptides - genetics; Peptides - metabolism; RNA Interference; RNA, Small Interfering - genetics; Stomach Neoplasms - genetics; Stomach Neoplasms - metabolism; Stomach Neoplasms - pathology; Transfection
• ISSN: 1671-0274

To investigate the changes in proliferation, invasiveness, clone sphere formation and chemosensitivity of human gastric cancer cell lines of KATO-III CD133(+) cells transfected with small interfering RNA (siRNA) against CD133 gene. CD133(+) cells of KATO-III cell lines were isolated by magnetic activated cell sorting (MACS). CD133 siRNA was designed and synthesized, and then transfected into KATO-III CD133(+) cells. Cell fluorescence counting under confocal laser scanning microscope was used to determine the transfection efficiency after transfection with the CD133 FITC-siRNA. The knock-down effect of the CD133 gene and expression of epithelial-mesenchymal transition (EMT)-related factors were detected by RT-PCR and Western blotting. Cell counting kit-8 assay (CCK-8), transwell chamber and colony sphere forming assay were performed to measure the variation of cell proliferative, invasive, colony formation viability and chemosensitivity to 5-FU after the above-mentioned treatment. The transfection efficiency w