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Downregulation of CD147 expression alters cytoskeleton architecture and inhibits gelatinase production and SAPK pathway in human hepatocellular carcinoma cells.

Qian AR, Zhang W, Cao JP, Yang PF, Gao X, Wang Z, Xu HY, Weng YY, Shang P - J. Exp. Clin. Cancer Res. (2008)

Bottom Line: Confocal microscopy was used to determine the effects of si-CD147 on SMMC-7721 cells' cytoskeleton.Western blot assay was utilized to detect the effects of si-CD147 on focal adhesion kinase (FAK), vinculiln and mitogen-activated protein kinase (MAPK) expression in SMMC-7721 cells.Moreover, the alteration of cell behavior may be related to SAPK/JNK Pathway. siRNA against CD147 may be a possible new approach for HCC gene therapy.

View Article: PubMed Central - HTML - PubMed

Affiliation: Key Laboratory for Space Biosciences & Biotechnology, Institute of Special Environmental Biophysics, Faculty of Life Sciences, Northwestern Polytechnical University, Xi'an 710072, PR China. qianair@nwpu.edu.cn

ABSTRACT

Background: CD147 plays a critical role in the invasive and metastatic activity of hepatocellular carcinoma (HCC) cells by stimulating the surrounding fibroblasts to express matrix metalloproteinases (MMPs). Tumor cells adhesion to extracellular matrix (ECM) proteins is the first step to the tumor metastasis. MMPs degrade the ECM to promote tumor metastasis. The aim of this study is to investigate the effects of small interfering RNA (siRNA) against CD147 (si-CD147) on hepatocellular carcinoma cells' (SMMC-7721) architecture and functions.

Methods: Flow cytometry and western blot assays were employed to detect the transfection efficiency of si-CD147. Confocal microscopy was used to determine the effects of si-CD147 on SMMC-7721 cells' cytoskeleton. Invasion assay, gelatin zymography and cell adhesion assay were employed to investigate the effects of si-CD147 on SMMC-7721 cells' invasion, gelatinase production and cell adhesive abilities. Western blot assay was utilized to detect the effects of si-CD147 on focal adhesion kinase (FAK), vinculiln and mitogen-activated protein kinase (MAPK) expression in SMMC-7721 cells.

Results: Downregulation of CD147 gene induced the alteration of SMMC-7721 cell cytoskeleton including actin, microtubule and vimentin filaments, and inhibited gelatinase production and expression, cells invasion, FAK and vinculin expression. si-CD147 also blocked SMMC-7721 cells adhesion to collagen IV and phosphorylation level of SAPK/JNKs. SAPK/JNKs inhibitor SP600125 inhibited gelatinase production and expression.

Conclusion: CD147 is required for normal tumor cell architecture and cell invasion. Downregulation of CD147 affects HCC cell structure and function. Moreover, the alteration of cell behavior may be related to SAPK/JNK Pathway. siRNA against CD147 may be a possible new approach for HCC gene therapy.

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Related in: MedlinePlus

Effects of si-CD147 RNA on SMMC-7721 cells' invasive abilities. The invasive abilities of SMMC-7721 cells transfected with negative control and si-CD147 RNA were examined by chamber assay. HE staining and quantitative analysis were employed to analyze the number of the transmembrane cells. Transmembrane cells number dramatically reduced in SMMC-7721 cells transfected with si-CD147 (A). The graph (B) compares the number of transmembrane cells and indicates an 85% inhibition by si-CD147. ***P < 0.001 v.s. control.
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Figure 3: Effects of si-CD147 RNA on SMMC-7721 cells' invasive abilities. The invasive abilities of SMMC-7721 cells transfected with negative control and si-CD147 RNA were examined by chamber assay. HE staining and quantitative analysis were employed to analyze the number of the transmembrane cells. Transmembrane cells number dramatically reduced in SMMC-7721 cells transfected with si-CD147 (A). The graph (B) compares the number of transmembrane cells and indicates an 85% inhibition by si-CD147. ***P < 0.001 v.s. control.

Mentions: In order to confirm whether CD147 is involved in the process of cell motility, we used invasion assay to determine the impact of siRNA-CD147 on SMMC-7721 cells' invasion. 5 × 105 cells transfected with si-CD147 or control siRNA were added to millicell inserts coated with matrigel. We observed that the number of infiltrating cells was significantly decreased in SMMC-7721 cells transfected with si-CD147 compared with that in control cells and the inhibitory rate was about 85% (Figure. 3A and 3B).


Downregulation of CD147 expression alters cytoskeleton architecture and inhibits gelatinase production and SAPK pathway in human hepatocellular carcinoma cells.

Qian AR, Zhang W, Cao JP, Yang PF, Gao X, Wang Z, Xu HY, Weng YY, Shang P - J. Exp. Clin. Cancer Res. (2008)

Effects of si-CD147 RNA on SMMC-7721 cells' invasive abilities. The invasive abilities of SMMC-7721 cells transfected with negative control and si-CD147 RNA were examined by chamber assay. HE staining and quantitative analysis were employed to analyze the number of the transmembrane cells. Transmembrane cells number dramatically reduced in SMMC-7721 cells transfected with si-CD147 (A). The graph (B) compares the number of transmembrane cells and indicates an 85% inhibition by si-CD147. ***P < 0.001 v.s. control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2572039&req=5

Figure 3: Effects of si-CD147 RNA on SMMC-7721 cells' invasive abilities. The invasive abilities of SMMC-7721 cells transfected with negative control and si-CD147 RNA were examined by chamber assay. HE staining and quantitative analysis were employed to analyze the number of the transmembrane cells. Transmembrane cells number dramatically reduced in SMMC-7721 cells transfected with si-CD147 (A). The graph (B) compares the number of transmembrane cells and indicates an 85% inhibition by si-CD147. ***P < 0.001 v.s. control.
Mentions: In order to confirm whether CD147 is involved in the process of cell motility, we used invasion assay to determine the impact of siRNA-CD147 on SMMC-7721 cells' invasion. 5 × 105 cells transfected with si-CD147 or control siRNA were added to millicell inserts coated with matrigel. We observed that the number of infiltrating cells was significantly decreased in SMMC-7721 cells transfected with si-CD147 compared with that in control cells and the inhibitory rate was about 85% (Figure. 3A and 3B).

Bottom Line: Confocal microscopy was used to determine the effects of si-CD147 on SMMC-7721 cells' cytoskeleton.Western blot assay was utilized to detect the effects of si-CD147 on focal adhesion kinase (FAK), vinculiln and mitogen-activated protein kinase (MAPK) expression in SMMC-7721 cells.Moreover, the alteration of cell behavior may be related to SAPK/JNK Pathway. siRNA against CD147 may be a possible new approach for HCC gene therapy.

View Article: PubMed Central - HTML - PubMed

Affiliation: Key Laboratory for Space Biosciences & Biotechnology, Institute of Special Environmental Biophysics, Faculty of Life Sciences, Northwestern Polytechnical University, Xi'an 710072, PR China. qianair@nwpu.edu.cn

ABSTRACT

Background: CD147 plays a critical role in the invasive and metastatic activity of hepatocellular carcinoma (HCC) cells by stimulating the surrounding fibroblasts to express matrix metalloproteinases (MMPs). Tumor cells adhesion to extracellular matrix (ECM) proteins is the first step to the tumor metastasis. MMPs degrade the ECM to promote tumor metastasis. The aim of this study is to investigate the effects of small interfering RNA (siRNA) against CD147 (si-CD147) on hepatocellular carcinoma cells' (SMMC-7721) architecture and functions.

Methods: Flow cytometry and western blot assays were employed to detect the transfection efficiency of si-CD147. Confocal microscopy was used to determine the effects of si-CD147 on SMMC-7721 cells' cytoskeleton. Invasion assay, gelatin zymography and cell adhesion assay were employed to investigate the effects of si-CD147 on SMMC-7721 cells' invasion, gelatinase production and cell adhesive abilities. Western blot assay was utilized to detect the effects of si-CD147 on focal adhesion kinase (FAK), vinculiln and mitogen-activated protein kinase (MAPK) expression in SMMC-7721 cells.

Results: Downregulation of CD147 gene induced the alteration of SMMC-7721 cell cytoskeleton including actin, microtubule and vimentin filaments, and inhibited gelatinase production and expression, cells invasion, FAK and vinculin expression. si-CD147 also blocked SMMC-7721 cells adhesion to collagen IV and phosphorylation level of SAPK/JNKs. SAPK/JNKs inhibitor SP600125 inhibited gelatinase production and expression.

Conclusion: CD147 is required for normal tumor cell architecture and cell invasion. Downregulation of CD147 affects HCC cell structure and function. Moreover, the alteration of cell behavior may be related to SAPK/JNK Pathway. siRNA against CD147 may be a possible new approach for HCC gene therapy.

Show MeSH
Related in: MedlinePlus