Effects of siRNA-targeting BMP-2 on the abilities of migration and invasion of human liver cancer SMMC7721 cells and its mechanism.
To observe the effects of small interfering RNA (siRNA)-targeting bone morphogenetic protein (BMP)-2 on the abilities of migration and invasion of human liver cancer SMMC7721 cells and its mechanism. Three siRNAs-targeting BMP-2 gene were synthesized. There were six groups including group I (non-transfected cells), group II (only liposome-transfected cells), group III (non-specific siRNA-transfected cells) and groups IV-VI (siRNA-A, siRNA-B and siRNA-C-targeting BMP-2 transfected cells, respectively). SMMC7721 cells were instantaneously transfected using lipofectamine method. The levels of mRNA and protein of BMP-2 in cells were determined with reverse transcription-PCR and western blotting. The abilities of migration and invasion of transfected cells were assessed using scratch test and in vitro invasion assay, respectively. The protein levels of p-ERK, p-JNK and p-p38 and the protein levels of MMP-2 and MMP-9 were evaluated with western blot 48 h after siRNA-B-targeting BMP-2 was transfected into liver cancer SMMC7721 cells. Expression of mRNA and protein of BMP-2 in groups IV-VI were significantly inhibited, especially in group V. Cell scratch width was significantly greater in group V than in group I and III (P<0.01). In vitro invasion assay suggested that the number of invasion of cells was significantly lower in group V than in group I and III (P<0.05). Western blot indicated that the level of p-ERK was significantly decreased (P<0.05), the levels of p-JHK and p-p38 were not significantly changed and the levels of MMP-2 and MMP-9 were significantly downregulated (P<0.05). siRNA-targeting BMP-2 can markedly inhibited the expression of BMP-2 in liver cancer SMMC7721 cells, and decrease the abilities of migration and invasion of liver cancer cells, especially siRNA-B. The inhibitory effects of siRNA-B-targeting BMP-2 on the abilities of migration and invasion of human liver cancer SMMC7721 cells may be caused by the downregulation of MMP-2 and MMP-9 through MAPK/ERK pathway, whereas is not related to MAPK/JNK and MAPK/p38 pathway.
Wu
J-B
JB
Department of Oncology, Second Affiliated Hospital, Nanchang University, China. kaishitengfei@163.com
Fu
H-Q
HQ
Huang
L-Z
LZ
Liu
A-W
AW
Zhang
J-X
JX
D015153:Blotting, Western; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics / Q000502:physiology*; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics* / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000235:genetics / Q000502:physiology
D055396:Bone Morphogenetic Protein 2; D034741:RNA, Small Interfering; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9