ABSTRACT
Objective:To investigate the expression pattern of MMP-11 and MMP-14 in breast carcinoma, and the effect of MMP-11 on breast carcinoma cell migration and invasion. Methods:MMP-11 and MMP-14 expression were examined in 161 invasive breast carcinoma tissue samples and 10 normal breast tissue samples. siRNA was used to knockout MMP-11 in breast carcinoma cell line MB-231 and Transwells were used to evaluate changes in migration ability and invasion ability. Results:Both MMP-11 and MMP-14 were highly expressed in breast carcinoma samples,122 and 149 samples out of 161,respectively. The expression of both proteins were correlated with lymph node metastasis and TNM staging. After knockout of MMP-11,the expression of both proteins decreased in MB-231 cell line and experiments show that the cell′s migration and invasion abilities were significantly weakened. Conclusion:MMP-11 and MMP-14 could promote invasion and metastasis of breast carcinoma. Knockout of MMP-11 results in the downregulated expression of MMP-14,and the inhibition of breast carcinoma cell′s migration and invasion. They could be potential prognostic markers and treatment targets for of breast carcinoma.
ABSTRACT
Background and purpose:A chemokine receptor,CXCR4,and its endogenous ligand,stromal cell-derived factor-1(SDF-1),have been recognized to be involved in the invasion and metastasis of cancer.Inhibition of CXCR4 may be a new therapeutic target.We studied whether shRNA-CXCR4 could inhibit the CXCR4 gene expression and the proliferation in MCF-7,MDA-MB-231 and MDA-MB-435s breast cancer cells.Methods:Through knockdown CXCR4 by shRNA,the CXCR4 mRNA expression was detected by RT-PCR and the CXCR4 protein expression was mesured by Western blot.The proliferation of breast cancer cells was evaluated by MTT and flow cytometry.Results:The CXCR4mRNA and its protein expression decreased significantly in MCF-7(the average level of CXCR4 mRNA and protein expression were 0.089,0.177 in PG-CXCR4 group,compared with 0.327 and 0.313 for mRNA expression,0.911,0.874 for protein expression in control and PG-HK group),MDA-MB-231(the average level of CXCR4 mRNA and protein expression were 0.152 and 0.153 in PG-CXCR4 group,compared with 0.40 and 0.45 for mRNA expression,0.829 and 0.878 for protein expression in control and PG-HK group)and MDA-MB-435s(the average level of CXCR4 mRNA and protein expression were 0.198 and 0.173 in PG-CXCR4 group,compared with 0.69 and 0.77 for mRNA expression,0.877 and 0.906 for protein expression in control and PG-HK group)breast cancer cells(P
ABSTRACT
Objective To study the effects of HYAL1 gene overexpression on invasive, angiogenic and proliferative ability of breast cancer cell lines MCF-7 and ZR-75-30. Methods Double-chamber co-culture technique was applied to construct the invasive model and angiogenic model in vitro, which was used to detect the invasive and angiogenic potential of breast cancer cell; MTT and flow cytometry were used to detect the proliferation of breast cancer cells. Results Breast cancer cells overexpressing HYAL1 gene showed stronger invasive potential and angiogenic potential than control cells, but had no significant difference on proliferative potential. Conclusion Overexpression of HYAL1 gene can promote the invasion and angiogenesis of breast cancer cells in vitro, but not affect the proliferation.
ABSTRACT
Objective To study whether shRNA-CXCR4 could inhibit the CXCR4 gene expression in MCF-7,MDA-MB-231 and MDA-MB-435s breast cancer cells.Methods After the knockdown of CXCR4 by shRNA,the CXCR4 mRNA expression by RT-PCR and the CXCR4 protein expression by Western blotting in breast cancer cells were examined.Results The CXCR4 mRNA expression and its protein expression were decreased significantly in MCF-7,MDA-MB-231 and MDA-MB-435s breast cancer cells by shRNA-CXCR4(P
ABSTRACT
Objective To study the effect of RNA interference (RNAi) on HYAL1 gene mRNA expression and the invasive potential of human breast cancer cell lines. Methods Chemically synthesized double stranded RNA (dsRNA) targeting HYAL1 was transfected into human breast cancer cell lines MDA-MB-231, MDA-MB453S, ZR-75 and ZR-75-30 using SiPORT Lipid. The transfection efficiency was observed under fluorescence confocal microscopy. Expression of HYAL1 mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR). Cell penetrate matrigel capacity were determined by in vitro experiment. Results HYAL1 -siRNA effectively inhibited HYAL1 mRNA expression ( P
ABSTRACT
ObjectiveTo construct the eukaryotic expression vector of human HYAL1 gene and obtain MCF-7 and ZR-75-30 cell clones expressing HYAL1 gene stably.MethodsThe cDNA encoding HYAL1 gene of human breast cancer was amplified by RT-PCR from the total RNA isolated from human MDA-MB-435S cells and inserted into pcDNA3.1/V5-His-TOPO vector.The recombinant plasmid was transferred into MCF-7 and ZR-75-30 cells.ResultsA 1332-bp DNA fragment was successfully amplified from human MDA-MB-435S cell.Restriction enzyme digestion analysis and DNA sequencing showed that HYAL1 gene was inserted into recombinant vector.RT-PCR analysis revealed that HYAL1 gene could be expressed stably in the transfected MCF-7 and ZR-75-30 and it had strong invasive potential.ConclusionThe eukaryotic expression vector of human HYAL1 gene was successfully constructed.MCF-7 and ZR-75-30 cell clones that can express HYAL1 gene were obtained and can promote the invasion.
ABSTRACT
Objective To study the expression of chemokine receptor-4(CXCR4) in human breast cancer and its correlation with the prognostic factors.Methods Forty-four samples of breast cancer were collected from Sep.2004 to Jan.2006.The expressions of CXCR4 mRNA and protein were evaluated by RT-PCR and Western blotting respectively.The expressions of CXCR4,estrogen receptor(ER),progesterone receptor(PR) and C-erbB-2 were examined by immunohistochemical methods.The clinicopathological features,such as ages,menstrual status,lymph node metastasis,tumor size and histological grading were recorded and analyzed.The relationship between CXCR4 and the clinicopathological features was analyzed.Results The immunohistochemical staining showed that CXCR4 expressed mainly in cytoplasm and little in nuclei.CXCR4 mRNA and protein expressed differently in 44 samples of breast cancer,and the expressive level increased with the increase in metastatic lymph nodes number.Compared with axillary lymph nodes metastasis negative group,the expression of CXCR4 mRNA and protein increased significantly in ≥4 and 1-3 nodes metastasis groups(P0.05).Conclusion CXCR4 might play an important role in promoting the metastasis of breast cancer.