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OBJECTIVE:To prepare argininate betulinic acid,and to investigate the effect of the proliferation of triple-negative human breast cancer cell MDA-MB-231. METHODS:By using argininate as the solubilization carrier,argininate betulinic acid was prepared by co-grinding equal molar ratio of betulinic acid and argininate. The argininate betulinic acid was characterized with powder X-ray diffractometry,infrared spectroscopy and differential scanning calorimetry. The solubility of betulinic acid and argininate betulinic acid were compared. MTT method was used to assay the effects of 15,30,60,120 μ g/mL betulinic acid, argininate betulinic acid and 5-FU on the proliferation of MDA-MB-231 cell. RESULTS:Prepared argininate betulinic acid was a new phase which was different from the physical mixing of argininate and betulinic acid,among which carboxyl group of betulinic acid and amino group of argininate formed as a salt,and the salt had no obvious melting peak. Betulinic acid was almost insoluble in water. The solubility of betulinic acid in argininate betulinic acid aqueous solution was 50.72 μg/mL. Compared with betulinic acid,the inhibitory rate of argininate betulinic acid on the growth of MDA-MB-231 cell was increased significantly(P<0.05), there was no statistical significance between its effect and 5-FU(P>0.05). CONCLUSIONS:Argininate betulinic acid with good solubility is prepared successfully,and can inhibit the proliferation of MDA-MB-231 cell.
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Objective To investigate the mechanism of xCT on tumor metastasis in breast cancer cell MDA-MB-231. Methods Wound scratch assay and Transwell assay were performed to evaluate the effect of disruption and knockdown of xCT on cell migration and cell invasion in breast cancer cell MDA-MB-231 .Western Blot and RT-PCR were used to detect the expression levels of autophagy and EMT related markers in breast cancer cell MDA-MB-231 after treatment with sulfasalazine (SASP), an inhibitor of xCT activity and SLC7A11-RNAi.Results Both the scratch assay and the transwell migration assay showed that inhibition of xCT reduced the motility of MDA-MB-231 .The expression level of autophagy related protein LC3-Ⅱ/LC3-Ⅰwas elevated, the protein level of transcription factor Snail was down-regulated, while the mRNA level of Snail did not change in xCT inhibited MDA-MB-231 cells compared with MDA-MB-231 cells.Epithelial marker E-cadherin was up-regulated but mesenchymal marker Vimentin was down-regulated when xCT was deficient.Con-clusion Our current studies show that xCT is an endogenous regulator of tumor growth and metastasis in MDA -MB-231 and the expression level of xCT determines the phenotypes of MDA-MB-231 cells in invasion and migration in vitro.Inhibition of xCT can activate autophagy , induce the degradation of Snail ,and attenuate the EMT process in highly metastatic MDA-MB-231 cells.
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Background and purpose:MicroRNA-340 (miR-340) has been demonstrated to play a role of negative regulation in many kinds of tumor, however, there are few reports about the relationship between miR-340 in proliferation and apoptosis of breast cancer cell. This study was aimed to explore the effect of miR-340 on proliferation and apoptosis of breast cancer cell MDA-MB231. Methods: The pre-miR-340 or anti-miR-340 were transiently transfected into breast cancer cell MDA-MB231 with LipofectamineTM2000. miR-340 level was detected by RT-PCR. The Western blot was performed to detect the protein level of cleaved-caspase-3. The inhibition rate of cell proliferation was evaluated by MTT assay. The cell apoptosis was studied by lfow cytometry. Results:The pre-miR-340 facilitated the expression of miR-340 in MDA-MB231 cells. The pre-miR-340 enhanced the protein level of cleaved-caspase-3, inhibited the proliferation of MDA-MB231 cells and increased its apoptosis. On the contrary, the expression of miR-340 was inhibited by anti-miR-340 in MDA-MB231 cells. The protein level of cleaved-caspase-3 was reduced after the anti-miR-340-transfected MDA-MB231 cells. Anti-miR-340 promoted the proliferation of MDA-MB231 cells. Decreased apoptosis of MDA-MB231 cells was observed by lfow cytometry. Conclusion:The overexpression of miR-340 can effectively inhibit the proliferation and increase the apoptosis of MDA-MB231 cells, which may be explained by up-regulating of protein cleaved-caspase-3 level.