This study examined the effect of CD24 on anoikis of ovarian cancer cells. The expres- sion of CD24 was detected by RT-PCR and Western blotting in ovarian cancer cells with high metas- tatic potential （HO-8910PM cell...This study examined the effect of CD24 on anoikis of ovarian cancer cells. The expres- sion of CD24 was detected by RT-PCR and Western blotting in ovarian cancer cells with high metas- tatic potential （HO-8910PM cells） and low metastatic potential （A2780 cells）. Cell viability and cell proliferation were detected by MTT assay in suspension culture and adhesion culture. Soft agar cul- ture was used to observe the colony formation. Anoikis was flow cytometrically detected. The results showed that the expression levels of CD24 mRNA and protein were significantly higher in HO-8910PM cells than in A2780 cells （P〈0.01）. In the suspension culture and soft agar culture, the HO-8910PM cells formed larger and more colonies （35.334-5.51 vs. 16.674-4.04; P〈0.01）, and showed a stronger resistance to anoikis than A2780 cells did （cell apoptosis rate： 5.93%4-2.38% vs. 16.32%-4-2.00%; P〈0.01）. After treated with CD24 monoclonal antibodies, the number of colony formed in HO-8910PM and A2780 cells was significantly decreased （9.334-2.52 and 8.004-2.00, re- spectively）, and the anoikis rate of the two cell lines was also markedly increased （23.11%4-2.87% and 28.36%~2.29%, respectively）. Our study suggested that CD24 may play an important role in the development of anoikis resistance and CD24 can be used as a new therapeutic target to induce anoikis and inhibit metastasis in ovarian cancer.展开更多
Anoikis is a form of apoptosis induced upon cell detachment from extracellular matrix. It has been determined that acquisition of resistance to anoikis is a critical step for tumor cell metastasis. MiR-21, the most pr...Anoikis is a form of apoptosis induced upon cell detachment from extracellular matrix. It has been determined that acquisition of resistance to anoikis is a critical step for tumor cell metastasis. MiR-21, the most prominent oncomiR, plays an important role in tumor progression. In this study, we revealed that up-regulation of miR-21 in human esophageal adenocarcinoma （EA） is associated with lymph node metastasis and poor survival rate. Because of the established anti-apoptosis effect of miR-21, it is tempting to speculate that miR-21 might contribute to tumor metastasis by regulating anoikis, qRT-PCR analysis demonstrated that miR-21 expression in OE33/AR cells （subpopulation of human EA OE33 cells that acquired resistance to anoikis） was significantly increased. Also, transfection of miR-21 mimics provided OE33 cells resisting to anoikis. By luciferase assays, we verified that PDCD4 and PTEN were the functional targets of miR-21. In mouse model, via tail vein injection experiment, we showed that the metastasis formation of OE33 cells in vivo could be mediated by changing the miR-21 expression pattern. Taken together, our findings suggested that miR-21 was involved in the regulation of anoikis in human EA cells. Targeting miR-21 may provide a novel strategy to prevent metastasis.展开更多
文摘This study examined the effect of CD24 on anoikis of ovarian cancer cells. The expres- sion of CD24 was detected by RT-PCR and Western blotting in ovarian cancer cells with high metas- tatic potential （HO-8910PM cells） and low metastatic potential （A2780 cells）. Cell viability and cell proliferation were detected by MTT assay in suspension culture and adhesion culture. Soft agar cul- ture was used to observe the colony formation. Anoikis was flow cytometrically detected. The results showed that the expression levels of CD24 mRNA and protein were significantly higher in HO-8910PM cells than in A2780 cells （P〈0.01）. In the suspension culture and soft agar culture, the HO-8910PM cells formed larger and more colonies （35.334-5.51 vs. 16.674-4.04; P〈0.01）, and showed a stronger resistance to anoikis than A2780 cells did （cell apoptosis rate： 5.93%4-2.38% vs. 16.32%-4-2.00%; P〈0.01）. After treated with CD24 monoclonal antibodies, the number of colony formed in HO-8910PM and A2780 cells was significantly decreased （9.334-2.52 and 8.004-2.00, re- spectively）, and the anoikis rate of the two cell lines was also markedly increased （23.11%4-2.87% and 28.36%~2.29%, respectively）. Our study suggested that CD24 may play an important role in the development of anoikis resistance and CD24 can be used as a new therapeutic target to induce anoikis and inhibit metastasis in ovarian cancer.
基金This project was supported by grants from the National Natural Science Foundation of China （No. 81470818, No. 81472735 and No. 81472033）.
文摘Anoikis is a form of apoptosis induced upon cell detachment from extracellular matrix. It has been determined that acquisition of resistance to anoikis is a critical step for tumor cell metastasis. MiR-21, the most prominent oncomiR, plays an important role in tumor progression. In this study, we revealed that up-regulation of miR-21 in human esophageal adenocarcinoma （EA） is associated with lymph node metastasis and poor survival rate. Because of the established anti-apoptosis effect of miR-21, it is tempting to speculate that miR-21 might contribute to tumor metastasis by regulating anoikis, qRT-PCR analysis demonstrated that miR-21 expression in OE33/AR cells （subpopulation of human EA OE33 cells that acquired resistance to anoikis） was significantly increased. Also, transfection of miR-21 mimics provided OE33 cells resisting to anoikis. By luciferase assays, we verified that PDCD4 and PTEN were the functional targets of miR-21. In mouse model, via tail vein injection experiment, we showed that the metastasis formation of OE33 cells in vivo could be mediated by changing the miR-21 expression pattern. Taken together, our findings suggested that miR-21 was involved in the regulation of anoikis in human EA cells. Targeting miR-21 may provide a novel strategy to prevent metastasis.
