Knocking down osteopontin expression by specific siRNA reduces the in vitro invasiveness of human hepatocellular carcinoma cells.
- Author:
Xiao-Qun ZHU
1
;
Qing-Hai YE
;
Ke-Feng LEI
;
Jie CHEN
;
Lun-Xiu QIN
Author Information
- Publication Type:Journal Article
- MeSH: Carcinoma, Hepatocellular; genetics; pathology; Cell Line, Tumor; Cell Movement; Cell Proliferation; Down-Regulation; Humans; Liver Neoplasms; genetics; pathology; Neoplasm Invasiveness; Osteopontin; genetics; RNA Interference; RNA, Messenger; biosynthesis; genetics; RNA, Small Interfering; genetics; Transfection
- From: Chinese Journal of Oncology 2006;28(6):404-407
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo study the effect of osteopontin (OPN) expression down-regulated by RNA interference (RNAi) on the invasiveness of hepatocelluar carcinoma cell line HCC-LM3.
METHODSHCC-LM3 cells were transfected with the chemically synthesized small interfering RNA (siRNA) formulated by lipofectamine 2000. Wild type HCC-LM3 and HCC-LM3 cells transfected with non-specific siRNA served as controls. Real-time PCR and Western blotting were used to quantify the mRNA and OPN protein levels. The malignant phenotypes of transfected HCC-LM3 cells including cellular growth rate, colony formation and Matrigel invasion activities were analyzed.
RESULTSSequence-specific siRNAs targeting OPN suppressed OPN RNA expression by 79% and also decreased OPN protein level by 81% in HCC-LM3 cells. The number of formed colonies and migrating numbers in vitro were decreased in HCC-LM3 cells transfected using sequence-specific siRNAs targeting OPN relative to controls (P < 0.05).
CONCLUSIONThis study demonstrated that specific siRNA is able to reduce OPN at both the mRNA and protein levels and significantly diminishes the invasiveness of hepatocellular carcinoma cells.
