ABSTRACT
Objective To investigate the effects of ER-α36 on invasion of human gastric cancer cell lines SGC7901 by miR-143. Methods Lentiviral vectors were constructed to generate up-and down-regulations of miR-143 lentiviruses (LV-miR-143 and LV-anti-miR-143,respectively).The viruses were used to infect human gastric cancer cell lines SGC7901.The ER-α36 protein expression level and the invasion of constructed cells were detected by Western blot and transwell. The target gene of miR-143 was predicted by bioinformatics tools.Luciferase reporter assay was carried out to confirm the predicted target gene. Results The infection efficiency of the lentivirus titers of LV-miR-143 and LV-anti-miR-143 were over 80% shown by the green fluorescence. The ER-α36 expression level,the cell invasion in LV-miR-21 group were significantly lower than those in LV-anti-miR-21 group(P<0.05). Conclusions miR-143 plays an important role in the negative control of gastric cancer invasion by the regulation of ER-α36.
ABSTRACT
Over-expression of Fas ligand (FasL) on tumor cell surface can induce the apoptosis of specific activated tumor infiltrating lymphocytes (TILs) via the Fas/FasL pathway, leading to the formation of a site of immune privilege surrounding the tumor mass for escaping immune surveillance and promoting tumor proliferation, invasion and metastasis. The blocking effect of miR-21 on FasL-mediated apoptosis in breast cancers was investigated in this study. The expression levels of miR-21 and FasL in human breast carcinoma cell lines were detected by using RT-PCR and Western blotting. FasL as a target gene of miR-21 was identified by Luciferase assay. The apoptosis of Jurkat T lymphocytes induced by MCF-7 cells was determined by flow cytometry. It was found that in four human breast cancer cell lines, FasL expression level in MCF-7 cells was the highest, while miR-21 was down-regulated the most notably. After miR-21 expression in MCF-7 cells was up-regulated, FasL was identified as a target gene of miR-21. When the effector/target (E/T) ratio of MCF-7 cells and Jurkat cells was 10:1, 5:1 and 1:1, the inhibitory rate of apoptosis of Jurkat T lymphocytes induced by MCF-7 cells was 95.81%, 93.16% and 91.94%, respectively. It is suggested that in breast cancers miR-21 expression is negatively associated with FasL expression, and FasL is a target gene of miR-21. miR-21 targeting and regulating FasL-mediated apoptosis will bring us the possibility of a new tumor immunotherapy via breaking tumor immune privilege.
Subject(s)
Female , Humans , Apoptosis , Genetics , Breast Neoplasms , Genetics , Fas Ligand Protein , Metabolism , Flow Cytometry , Gene Expression Regulation, Neoplastic , MCF-7 Cells , MicroRNAs , Genetics , Signal TransductionABSTRACT
Over-expression of Fas ligand (FasL) on tumor cell surface can induce the apoptosis of specific activated tumor infiltrating lymphocytes (TILs) via the Fas/FasL pathway, leading to the formation of a site of immune privilege surrounding the tumor mass for escaping immune surveillance and promoting tumor proliferation, invasion and metastasis. The blocking effect of miR-21 on FasL-mediated apoptosis in breast cancers was investigated in this study. The expression levels of miR-21 and FasL in human breast carcinoma cell lines were detected by using RT-PCR and Western blotting. FasL as a target gene of miR-21 was identified by Luciferase assay. The apoptosis of Jurkat T lymphocytes induced by MCF-7 cells was determined by flow cytometry. It was found that in four human breast cancer cell lines, FasL expression level in MCF-7 cells was the highest, while miR-21 was down-regulated the most notably. After miR-21 expression in MCF-7 cells was up-regulated, FasL was identified as a target gene of miR-21. When the effector/target (E/T) ratio of MCF-7 cells and Jurkat cells was 10:1, 5:1 and 1:1, the inhibitory rate of apoptosis of Jurkat T lymphocytes induced by MCF-7 cells was 95.81%, 93.16% and 91.94%, respectively. It is suggested that in breast cancers miR-21 expression is negatively associated with FasL expression, and FasL is a target gene of miR-21. miR-21 targeting and regulating FasL-mediated apoptosis will bring us the possibility of a new tumor immunotherapy via breaking tumor immune privilege.
ABSTRACT
OBJECTIVE@#Using computer image-analyze technique (CIAT) to study changes of geometry parameters in human spleen nuclei and seek a new experimental method to deduce the estimation the postmortem interval (PMI).@*METHODS@#31 cadavers that known accurate PMI, sampled and smeared respectively every hour within the first 36 hours after death, fixed with cold Carony fixation, stained by Feulgen-van's method, and measured 5 geometry parameters using the image-analyze instrument including Area (A), Mean-Dia (MD), Average Diameter (AD), perimeter (P), Index of density (ID).@*RESULTS@#A, MD, AD and P in the human spleen nuclei have no correlation with the PMI. But ID rose regularly with the prolongation of PMI in 36 hours. There was a definite correlation between ID and the PMI, r=0.983, linear regression equation with PMI (hours) as the dependent variable was calculated for ID.@*CONCLUSION@#Geometry parameter ID was proved to be preferable indexes for estimation of PMI in 36 hours.