RESUMO
Purpose To investigate the effect of specific long-chain non-coding RNA AP000344. 3 on the proliferation and invasion of bladder cancer cells and its mechanism. Methods qRT-PCR was used to detect the expression of AP000344. 3 in bladder cancer cells and normal bladder epithelial cells. The cancer cells with the lowest expression rate were selected for subsequent experiments. The AP000344. 3 plasmid or the negative control plasmid was transferred into bladder cancer cells by Lipofectamine 2000, and the transfection efficiency of AP000344. 3 was detected by qRT-PCR. Cell proliferation activity was measured by MTT method,and cell invasion ability was detected by Transwell assay. Bioinformatics was used to predict downstream miRNA and downstream genes of AP000344. 3. qRT-PCR and Western blot were used to detect the expression of downstream genes. Results The expression of AP000344. 3 in bladder cancer cells was significantly lower than that in normal bladder epithelial cells (P < 0. 01) ,and the expression of AP000344. 3 was the lowest in BIU87 cells (P < 0. 01) . The expression of AP000344. 3 in BIU87 cells was up-regulated at 48 h after transfection with AP000344. 3 (P < 0. 01) . The proliferation activity of BIU87 cells was decreased (P < 0. 05) ,and the cell invasion ability was decreased (P < 0. 05) . AP000344. 3 can target and bind to miR-135a-5p,and miR-135a-5p to human chemokine-like factor superfamily member 3 (CMTM3) . After up-regulation of AP000344. 3,miR-135a-5p expression was down-regulated (P < 0. 01) ,and CMTM3 was up-regulated in mRNA and protein expression (P < 0. 01) . Conclusion AP000344. 3 is significantly down-regulated in bladder cancer cells,and up-regulation of AP000344. 3 can inhibit the proliferation and invasion of bladder cancer cells. The mechanism may be to inhibit the expression of miR-135a-5p and up-regulate the expression of CMTM3 protein,providing a theoretical basis for finding new therapeutic targets for bladder cancer.
RESUMO
Purpose To observe the effect of specific miR-448 inhibitor on the proliferation and migration of prostate cancer cells and its molecular mechanism. Methods Real-time fluo-rescent quantitative polymerase chain reaction ( qRT-PCR) was used to detect the expression of miR-448 in normal prostate epi-thelial cells and cancer cells. The cells with the highest expres-sion of miR-448 were selected for follow-up experiment. The miR-448 inhibitor or miR-NC was transferred into prostate canc-er cells using liposome transfection reagent. The expression of miR-448 and CMTM3 mRNA were detected by qRT-PCR. The expression of related proteins were detected by Western blotting. MTT assay was used to detect the cell proliferation activity. Tr-answell assay was used to detect the cell migration ability. Re-sults The expression of miR-448 in normal prostate epithelial cells was significantly lower than that in cancer cells ( P ＜0. 01), and the expression of miR-448 was the highest in DU- 145 cells ( P ＜0. 01). The expression of miR-448 in DU-145 cells was down-regulated 48 h after transfection with miR-448 in-hibitor (P＜0. 01). The expression of CMTM3 mRNA was up-regulated (P＜0. 01). The expression of CMTM3, E-cadherin and β-catenin proteins were up-regulated. The expression of N-cadherin and Snail proteins were down-regulated. Cell prolifera-tion was decreased (P＜0. 05). Cell migration ability decreased (P ＜ 0. 01 ). Conclusion miR-448 is highly expressed in prostate cancer cells. miR-448 inhibitor can down-regulate the expression of miR-448 in DU-145 cells, up-regulate the expres-sion of CMTM3 protein, inhibit the proliferation and migration of prostate cancer cells, which showing a potential function in mo-lecular therapy of prostate cancer.