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Objective To explore changes of invasion capability of cancer stem cells loading miR30a derived from enlarging and proliferative axillary lymph node of patients with breast cancer in nude mice.Methods MiR30a oligonucleotide fragment loaded by adenovirus vector was transfected into breast cancer stem cells isolated from enlarging and proliferative axillary lymphnodes of patients with breast cancer,MDA-MB-231 cell lines as the control.The cancer cells were injected into axillary subcutaneous fat of the nude mice for several weeks,and then,the expression of Vimentin or N-Cadherin in tumor tissues of each group of the mice was determined by immunohistochemistry and western blot.Results The average transfection rate of cancer stem cells loaded by miR30a was 62.5％,while it was 78.2％ for MDA-MB-231.The tumor volume was larger in adenovirus vector groups or in control groups of nude mice than in experimental groups induced by mir30a.Vimentin or N-cadherin in tumor tissues was significantly downexpressed in experimental groups with mir30a ((13.1±1.7)％,(15.3％±2.1)％)compared with that in adenovirus vector groups or in control groups((21.1±1.4)％,(25.3±1.6)％,P＜0.05),respectively.The difference between adenovirus vector groups and in control groups had no significant difference (P＞0.05).After inoculated for 6 weeks,except the subcutaneous plantations,no distant metastasis in nude mice was found.MDA-MB-231 cell lines groups had similar results.Conclusion The proliferative and invasive capability of cancer stem cells can be inhibited by miR30a,suggesting a new therapy for breast cancer.
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Objective The aims of this study were to investigate the expression of microRNA-30a(miR-30a)in human bladder cancer cell lines and their effects on the proliferation,apoptosis and migration of human bladder cancer cells.Methods The expression levels of miR-30a in bladder cancer cell lines(5637 and T24)and bladder epithelial immortalized cells(SV-HUC-1)were detected by real-time quantitative PCR(qRT-PCR).The expression of miR-30a was up-regulated or down-regulated by T24 cells transfected with miR-30a mimic or 5637 cells transfected with miR-30a inhibitors and controls using NC mimic or NC inhibitor.The effects of miR-30a expression on the proliferation,apoptosis and invasion of bladder cancer cells were investigated by flow cytometry,MTT and Transwell assays.Results The expression level of miR-30a in two bladder cancer T24 and 5637 cell lines was significantly lower than that in normal bladder SV-HUC-1 cell line(P<0.05),and the expression level of miR-30a was lower in the high degree of malignancy in bladder cancer T24 cells than that in malignant degree of relatively low 5637 cells.After 72h transfection,the values of optical density(OD)in the miR-30a mimic group(0.83±0.09)was significantly lower than that in NC mimic group(1.21±0.12)in T24 cells(P<0.01).The OD values of miR-30a inhibitor group(1.28±0.14)was significantly lower than that in the NC inhibitor group(1.09±0.14)in 5637 cells(P<0.01).The apoptotic rate of miR-30a mimic group in T24 cells(21.27±2.42)% was significantly higher than that in the NC mimic group(10.61±1.29)%(P<0.01).The apoptotic rate of the miR-30a inhibitor group in 5637 cells(6.78±2.57)% was significantly lower than that in the NC mimic group(13.42±1.40)%(P<0.01).The number of transmembrane cells in miR-30a mimic group in T24 cells(183.57±16.61)was significantly lower than that in NC mimic group(465.80±9.20)(P<0.01).The number of transmembrane cells in the miR-30a inhibitor group in 5637 cells(581.25±11.02)was significantly lower than that in NC mimic group(397.13±7.57)(P<0.01).Conclusion Up-regulation of miR-30a can inhibit the proliferation of bladder cancer cells,promote cell apoptosis and reduce the ability of migration and invasion in bladder cancer cells.The low expression of miR-30a in bladder cancer cells may be related to the development and metastasis in bladder cancer.
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Objective:To investigate the expression and clinical significance of microRNA-30a in renal cell carcinoma (RCC). Methods:The miRNA-30a expression in renal tissues and cell lines was detected using quantitative real-time polymerase chain reac-tion (qRT-PCR), and the relationship between miRNA-30a expression and the clinicopathological features of RCC was analyzed. The effect of miRNA-30a on cancer cells was evaluated using methyl thiazolyl tetrazolium (MTT) assay. In addition, a bioinformatics algo-rithm was adopted to predict the potential targets of miRNA-30a. Results:miRNA-30a was downregulated both in RCC tissues and cell lines. Patients with higher miRNA-30a expression exhibited better prognosis than those with lower miRNA-30a expression. Bioin-formatics algorithm and qRT-PCR both indicated that metadherin (MTDH) was a target of miRNA-30a. Moreover, MTT results showed that miRNA-30a could inhibit cell proliferation. Conclusion:miRNA-30a was inhibited in renal cancer, and low miRNA-30a expres-sion was associated with poor prognosis. In addition, miRNA-30a could inhibit the growth of cells through MTDH.