Knockdown of CD-74 in the Proliferative and Apoptotic Activity of Breast Cancer Cells
Hussain Al SsadhWaleed Al AbdulmonemZafar RasheedInamul Hasan MadarJamila AlhoderiSamah K Nasr EldeenAli AlradhwanNoura AlasmaelAbdullah S. AlkhamissNelson Fernández
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Abstract:
The cluster of differentiation (CD) 74 is known for its immunological functions and its elevated level was reported in various cancer cells.The aim of the present study was to investigate the expression and potential roles of CD74 in the proliferative and apoptotic activity of breast cancer.Expression of CD74, macrophage migration inhibitory factor (MIF) and CD44 was assayed in CAMA-1 and MDA-MB-231 cell lines using flow cytometry. CD74 was knocked down using CD74 siRNA-transfection in CAMA-1, and MDA-MB-231 cells and proliferation and apoptosis were determined in the transfected breast cancer cells.The data showed that CD74, MIF and CD44 were expressed in breast cancer cell lines and were associated with cell proliferation and apoptosis. Correlation analysis revealed that CD74 was positively correlated and colocalised with MIF on the cell-surface of CAMA-1 and MDA-MB-231. The knockdown of CD74 significantly reduced CAMA-1 and MDA-MB-231 cell proliferation and increased the level of apoptotic cells.We concluded that the interactions of CD74 with MIF and CD74 with CD44 could be a potential tumour marker for breast cancer cells. Moreover, the level of co-expression of MIF and CD74 or CD44 could be a surrogate marker for the efficacy of anti-angiogenic drugs, particularly in breast cancer tumours. In short, the study revealed the potential roles of CD74 in the proliferation and apoptosis of breast cancer which may serve as a potential therapeutic target for breast cancer.Keywords:
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Objective To investigate the mechanism underlying the anticancer activity of astragalus on human laryngeal cancer.Methods Hep-2 cells were treated with different concentrations of astragalus for 24 h.MTT assay was used to evaluate cell proliferation.Flow cytometry with PI staining and fluorescent microscopy with hoechst 33258 staining were used to estimate cell cycle distribution and cell apoptosis.Results Astragalus inhibited cellular proliferation in a dose dependent manner(P0.05).Flow cytometry analysis showed that treatment with astragalus resulted in accumulation of cells at the G2/M phase of the cell cycle and cell apoptosis in a dose dependent manner(P0.05).Marked apoptotic changes were observed by 33258 staining.Conclusion Astragalus inhibited cell proliferation and induced apoptosis of Hep-2 cells by regulating cell cycle,forming G2/M phase inhibition.
Astragalus
MTT assay
Cytometry
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Objective To investigate the effect of allicin on cell growth,cell apoptosis and cell cycle in human hepatoma cell HepG2 in vitro. Methods The human hepatoma cell HepG2 was cultured and treated by allicin in different concentrations in vitro. MTT assay was used to test dynamically the inhibitive effect of the cell growth. Morphologic change was observed by using the fluorescent microscope. The modulation of DNA cell cycle and apoptosis induction were measured by flow cytometry. Results Allicin had inhibitive effects on growth of HepG2 cells in a dose and time-dependent manner,with the IC50 value of(48.26±1.66)μg/mL,(31.89±1.51)μg/mL and(23.79±1.32)μg/mL at 24h,48h and 72h respectively. When the cells were treated with allicin at the concentration of 32μg/mL for 48h,HepG2 cells were notably induced into apoptosis. Flow cytometry showed that allicin could block cell growth at G2/M phase. Conclusion It is suggested that allicin inhibits HepG2 cell proliferation,which is associated with the induction of apoptosis and arrestment of cell cycle.
MTT assay
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Objective To study the effects of nm23-h1 on cell proliferation and possible mechanism in oral squamous cell carcinoma.Methods The expression of nm23-h1 was detected in six oral squamous cell carcinoma cell lines.The cell line of the lowest nm23-h1 expression with highest transfection efficiency was used to transfect nm23-h1 instantaneonsly.Cell proliferation was detected by the method of MTT and the expression of PKC-α examined by realtime PCR.Results High metastatic cell lines Tb and Tca8113M expressed the lowest nm23-h1 in six oral squamous cell carcinoma cell lines which involved Tca8113,Tb,Tca8113M,TSCC,OSC-4,Tca8113/CDDP.Attenuated cell proliferation and higher expression of PKC-α were oberseved in transfected Tb cells.Conclusion Transfection of nm23-h1 attenuated cell proliferation of Tb and PKC-α signaling might be involved in this process.
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Objective To observe the effect of miR-204 over-expression on growth and apoptosis of gastric cancer cells.Methods Expression of miR-204 in gastric cancer cells was detected by real time PCR and transfected into gastric cancer cells.Growth and apoptosis of gastric cancer cells were detected by MTS and flow cytometry,respectively.Results The expression level of miR-204 was low in gastric cancer cells,which was similar to that in gastric tissue.No significant change was observed in the growth and apoptosis of gastric cancer cells when miR-204 was over-expressed.Conclusion Over-expression of miR-204 has no significant effect on the growth and apoptosis of gastric cancer cells.
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Objective To explore the effect of Tanshinone ⅡA on cell proliferation,apoptosis and cell cycle of fibroblasts derived from keloid.Methods Fibroblasts derived from keloid were cultured with different concentration of Tanshinone ⅡA(0 μg/mL,50 μg/mL,100 μg/mL and 200 μg/mL) in vitro.CCK-8 was used to detect cell proliferation,and flow cytometry was used to analyze cell apoptosis,cell cycle at the different time.Results Cell proliferation of fibroblasts derived from keloid was decreased,apoptosis at the early phase was increased,and cell cycle in G0/G1 phase was arrested by different concentration of Tanshinone ⅡA with different intervention time(P0.001),especially in 200 μg/mL group.Variance analysis showed that those differences have statistical significance.Conclusion Tanshinon ⅡA can suppress cell proliferation,induce cell apoptosis and arrest cell cycle of fibroblasts derived from keloid.The effect of Tanshinone ⅡA was affected by different concentration and different intervention time.
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ObjectiveTo observe the effects of hPOT1 overexpression on cell cycle and apoptosis of HeLa cells. MethodsBy using an eukaryotic expression plasmid of pcDNA-hPOT1, liposomal mediates transient transfection of HeLa cells. The expressing effects of foreign genes were detected by RT-PCR and EMSA method. The cell cycles were analyzed by flow cytometry. And cell apoptosis was detected by Hoechst 33342 fluorescent staining. ResultsAfter 48 hours of the pcDNA3-hPOT1 transfected into HeLa cells, mRNA and protein analysis showed that hPOT1 could be efficiently expressed. Cell cycle in HeLa cells was blocked in S phase, and the overexpression of hPOT1 didnt show the effects on apoptosis. ConclusionThose results proved that hPOT1 functioned in the regulation of cell cycle in eukaryotic cells but not in cell apoptosis.
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Objective To investigate the effect of Quercetin on the proliferation and apoptosis of the colon carcinoma cell line Lovo.Methods The cultured Lovo cells were treated with different concentrations of Quercetin for different time.The proliferation of Lovo cells was detected by MTT method.Cell apoptosis and cell circle were detected by flow cytometry.Results Quercetin inhibited the proliferation of Lovo cells in a concentration-and time dependent manner.Quercetininduced the apoptosis and inhibted the cell circle of Lovo cells in a concentration-dependent manner.Conclusion Quercetin can induce the apoptosis of the colon carcinoma cell line Lovo,reduce the proliferation of Bcl-2,and inhibite the cell circle of colon carcinoma.
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[Objective] To study the effect of PERICARPIUM ZANTHOXYLI extracts on proliferation and apoptosis of SGC-7901 cell.[Method] The SGC-7901 cell was cultured in vitro.The inhibitory effect of PERICARPIUM ZANTHOXYLI extracts was determined by MTT;cell morphology was observed under optical microscope and electron microscope;cell apoptosis was detected by flow cytometry.[Result] PERICARPIUM ZANTHOXYLI extracts showed significant inhibitory effect to SGC-7901 cell,and the inhibitory effect was increased with the increase of concentration and time.The cell changed greatly in apoptosis characteristics.[Conclusion] PERICARPIUM ZANTHOXYLI extracts had potential anti-tumor value.
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Objective MiRNA is an important regulating factor in tumors. Our previous findings showed that the expression of miR-199a-5p was significantly down-regulated in clear-cell renal-cell carcinoma( CCRCC),suggesting an inhibiting effect of miR-199a-5p on tumor growth. This study aims to investigate the role of miR-199a-5p in the proliferation,apoptosis,invasion and cell cycle of CCRCC 786-0 cells. Methods Using liposome-mediated gene transfection,we transfected human CCRCC 786-0 cells with miR-199a-5p mimics( experimental group) and negative control. Then we detected the proliferation of the transfected cells by cell counting kit 8,their invasion ability by transwell assay,and their apoptosis and cell cycles by flow cytometry. Results Compared with the negative control,the 786-0 cells showed significantly increased proliferation ability( P 0. 05) and reduced invasion ability( P 0. 01) in the experimental group,but no obvious differences in apoptosis and cell cycles. Conclusion miR-199a-5p is capable of weakening the invasion ability of human CCRCC 786-0 cells in vitro.
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Objective To investigate the effects of SH3GL2 on apoptosis,proliferation and sensitivity to cis-diamminedichloroplatinum (DDP)in Hep2 cell. Methods After pcDNA3.1-SH3GL2 was constructed and transfected into Hep2 cell,the proliferation of the transfected cells was determined by methyl thiazolyl tetrazolium (MTT)and the apoptosis by flow cytometry with the treatment of DDP. Results The expression of SH3GL2 was increased after Hep2 cells were transfected with pcDNA3.1-SH3GL2,which was successfully constructed. Compared with control,the proliferation of transfectant cells was inhibited significantly (P 0.05),and significant increase of their apoptosis was observed(P 0.05). Compared with DDP-treated group,the proliferation was more significantly inhibited(P 0.05),and more significant increase of apoptosis was found in pcDNA3.1-SH3GL2-transfected cells (P 0.05). Conclusion SH3GL2 gene can inhibit proliferation and increase apoptosis of Hep2 cell,and can increase the sensitivity to DDP.
MTT assay
Growth inhibition
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