Lentivirus-mediated RNA interference of CD133 inhibits the proliferation of CD133(+) liver cancer stem cells and increases their cisplatin chemosensitivity / 南方医科大学学报
Journal of Southern Medical University
;
(12): 1741-1747, 2012.
Article
in Chinese
| WPRIM
| ID: wpr-352343
ABSTRACT
<p><b>OBJECTIVE</b>To study the effects of CD133 suppression by lentivirus-mediated RNA interference (RNAi) on the proliferation and chemosensitivity of CD133(+) cancer stem cells (CSCs) sorted from HepG2 cell line.</p><p><b>METHODS</b>CD133(+) and CD133- cells were sorted from HepG2 cell line by flow cytometry, and the expression of CD133 before and after cell sorting were detected. The stem cell property of sorted CD133(+) cells were validated by sphere-forming assay in vitro and xenograft experiments in vivo. Lentivirus-mediated short hairpin RNA (shRNA) targeting CD133 were transfected into CD133(+) cells, and CD133 mRNA and protein expressions of the transfected cells were detected by RT-PCR and Western blotting, respectively. Before and after the transfection, the proliferative ability of CD133(+) cells was evaluated by colony formation assay, and the cell growth inhibition rate and apoptosis following cisplatin exposure were detected using CCK-8 assay and flow cytometry.</p><p><b>RESULTS</b>The sorted CD133(+) cells showed a high purity of (88.74∓3.19)%, as compared with the purity of (3.36∓1.80)% before cell sorting. CD133(+) cells showed a high tumor sphere formation ability and tumorigenesis capacity compared with CD133- cells. CD133 shRNA transfection significantly inhibited CD133 mRNA and protein expressions in CD133(+) cells (P<0.01), resulting also in a significantly lowered cell proliferative ability (P<0.01) and an increased growth inhibition rate (P<0.01) and obviously increased cell apoptosis (P<0.05) after cisplatin exposure.</p><p><b>CONCLUSION</b>Lentivirus-mediated RNAi for CD133 suppression inhibits the proliferation of CD133(+) liver cancer stem cells and increases their chemosensitivity to cisplatin.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Peptides
/
Pharmacology
/
Neoplastic Stem Cells
/
Glycoproteins
/
Antigens, CD
/
Cisplatin
/
Lentivirus
/
Drug Resistance, Neoplasm
/
Cell Biology
/
RNA Interference
Limits:
Humans
Language:
Chinese
Journal:
Journal of Southern Medical University
Year:
2012
Type:
Article
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