RESUMEN
@#[摘 要] 目的：探讨miR-202-5p对口腔鳞状细胞癌（oral squamous carcinoma，OSCC）细胞生长、集落形成、迁移和侵袭的影响及其可能的机制。方法：通过qPCR法检测OSCC细胞系（Tca8113和SCC-4）和口腔角质细胞HOK中miR-202-5p和T细胞核因子c3（nuclear factor of activated T-cells isoform c3，NFATc3）mRNA的表达水平；将miR-202-5p mimic或/和NFATc3过表达质粒转染入Tca8113和SCC-4细胞，用MTT和集落形成实验检测转染对细胞增殖的影响，划痕伤口愈合实验和Transwell实验检测转染对细胞迁移和侵袭的影响，用Western blotting实验检测转染对NFATc3蛋白表达的影响；通过双荧光素酶报告基因实验验证miR-202-5p对候选靶基因NFATc3的直接调控作用。结果：与正常口腔角质细胞HOK相比，miR-202-5p在OSCC细胞Tca8113和SCC-4中呈低表达（均P<0.01），NFATc3 mRNA和蛋白质表达显著升高（P<0.01）。在Tca8113细胞和SCC-4细胞中，过表达miR-202-5p可显著抑制细胞生长、集落形成、迁移、侵袭以及细胞中NFATc3表达（均P<0.01）。NFATc3被证实是miR-202-5p的靶基因，过表达NFATc3能逆转miR-202-5p对OSCC细胞生长、迁移和侵袭的抑制作用。结论：miR-202-5p通过下调NFATc3表达发挥其肿瘤抑制功能，导致OSCC细胞的生长、迁移和侵袭受到抑制。
RESUMEN
Background: Long non-coding RNA (lncRNA) could participate in the process of tumorigenesis by regulating the expression of miRNA, but the molecular mechanism of lncRNA in the development and metastasis of colorectal cancer has not been elucidated. Aims: To investigate the molecular mechanism of lncRNA SOX21-AS1 affecting the biological process of colorectal cancer cells by regulating the expression of microRNA-202-5p (miR-202-5p). Methods: The colorectal cancer HCT116 and SW480 cells were transfected with SOX21-AS1 small interfering RNA (si-SOX21-AS1), miR-202-5p mimics and its inhibitor. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the mRNA expressions of SOX21-AS1 and miR-202-5p. Cell proliferation was detected by MTT assay. Flow cytometry was used to detect cell apoptosis. Transwell experiments were used to detect cell migration and invasion. Dual luciferase reporter assay was used to validate the targeted regulatory relationship between SOX21-AS1 and miR-202-5p. Western blotting was used to detect the protein expressions of cyclin D1, MMP-2 and cleaved caspase-3. Results: The mRNA expression of SOX21-AS1 in colorectal cancer cells was significantly higher than that in normal colonic mucosal epithelial cells (P<0.05), while the mRNA expression of miR-202-5p was significantly decreased (P<0.05). Silencing SOX21-AS1 or up-regulating miR-202-5p significantly inhibited proliferation, migration and invasion of HCT116, SW480 cells, the cell apoptosis rate was significantly increased (P<0.05), and significantly inhibited the protein expressions of cyclin D1 and MMP-2 (P<0.05), however, the protein expression of cleaved caspase-3 was significantly increased (P<0.05). SOX21-AS1 could bind to miR-202-5p and negatively regulate the expression and activity of miR-202-5p. Inhibition of miR-202-5p expression reversed the effect of silencing SOX21-AS1 on proliferation, apoptosis, migration and invasion of colorectal cancer cells. Conclusions: Silencing SOX21-AS1 inhibits the proliferation, migration and invasion of colorectal cancer cells and induces cell apoptosis by up-regulating the expression of miR-202-5p.
RESUMEN
Objective To study the value of miR-202-3p detection in the diagnosis and treatment of systemic lupus erythematosus (SLE) patients.Methods Two hundred and fifty cases of SLE and 100 cases of healthy controls from January 2017 to December 2018 were involved in the study.The expression of serum miR-202-3p in the 2 groups was detected by real-time quantitative polymerase chain reaction method,and its association with the clinicopathological features was analyzed.Statistical analyses were performed using t/Z-test,one-way analysis of variance,Pearson correlation,and receiver operating characteristic (ROC) curve.Results Compared with that in the control group (26.30±0.43),RA group (25.59±0.38)],the expression level of serum miR-202-3p in the SLE group (9.84±0.46) was significantly decreased (F=320.5,P＜0.01).The expression was lower in patients with active SLE (2.10±0.140) than that in those with stable SLE(14.67±0.39) and the difference was statistically significant (t =24.864,P＜0.01).The expression of miR-202-3p was negatively correlated with disease activity in systemic lupus erythematosus (SLEDAI) (r=-0.809 3,P＜0.01).The expression of miR-202-3p in patients with lupus nephritis combined with SLE (0.74±0.06) was significantly lower than that in patients without nephritis (2.81 ±0.15) and the difference was statistically significant (t=9.991,P＜0.01).The area under the ROC curve of serum miR-202-3p as a diagnosis of SLE was 0.974 [95％CI(0.955,0.988),P＜0.01],and its sensitivity and specificity was 90％ and 96.4％,respectively.Conclusion Serum miR-202-3p is highly expressed in SLE patients and is related to disease activity and renal injury in SLE patients.miR-202-3p may be used for SLE diagnosis.
RESUMEN
Objective To explore the effect and molecular mechanism of miR-202 on the differentiation of 3T3-L1 preadipocyte .Methods Through lentivirus infected with 3T3-L1 preadipocytes , we set up the AMO-miR-202 group and the random control group , then, these cells were induced to differentiate , nine days later, differentiation was assessed by Oil Red O staining and we examined the mRNA expression of PPARγ2 and aP2 by RT-PCR method. We examined the mRNA expression of PPARγ2,aP2 and PGC1βby Western blot method .Results After packa-ging lentivirus with AMO-miR-202 and random sequence control miRNA through cell line 293T, 80%-90%cells with fluorescence were found under fluorescence microscope; After these two lentivirus respectively infected with 3T3-L1 preadipocytes, About 70%-80%cells with fluorescence were found under fluorescence microscope .Oil Red O staining test showed that these cells with Oil Red O stained bright red fat droplets of AMO-miR-202 group and PPARγ2 and aP2 mRNA expression in the AMO-miR-202 group significantly lower than control groups (P<0.05). Western blot assay showed that the protein expression of PGC 1βin the AMO-miRNA-202 group was significantly increased(P<0.05), but the expression of aP 2 and PPARγ2 was significantly decreased (P<0.01).However, the random control group and the adipocyte group had no significant effect on the above indexes .Conclusions miR-202 can promote the differentiation of 3T3-L1 preadipocyte by inhibiting the protein expression of PGC 1βand im-proving the protein expression of PPARγ2 and aP2.
RESUMEN
Objective:To study the effects of the overexpression of hsa-miR-202 on the proliferation and molecular mechanism of lung cancer A549 cells. Methods:A sequence of hsa-miR-202 with ppG/miR/eGFP/Blasitidin pasmid was directionally connected and a eukaryotic expression vector pmiR-202 of the target hsa-miR-202 gene was constructed. pmiR-202 was transtected to A549 cell with Lipofectamine 2000. The WST assay was used to detect the cell proliferation rate, and RT-PCR was used to detect the relative gene expression levels. Western blot analysis was used to detect the IL-10 protein expression levels. The interaction between miR-202 and IL-10 was examined using a luciferase reporter assay. Results:The design from the DNA sequencing results shows that a eukaryot-ic expression vector of miR-202 was successfully constructed. The proliferation inhibition rates of A549 cells by Pmir-202 were 12%, 38%, and 52%. The differences in the treatment group compared with the blank control and negative control groups were statistically significant. The RT-PCR results showed that the relative expression levels of miR-202 increased after transfection with pmiR-202. Over-expression of miR-202 can downregulate the relative gene and protein expression levels of IL-10, and the relative levels were 25%and 0.75, respectively. Compared with the blank control and the negative control groups, the difference was statistically significant (P<0.05). The fluorescent activity was reduced when transfection was performed with miR-202 mimics, and IL-10-3'UTR plasmid was cloned. Conclusion: pmiR-202 effectively inhibited the proliferation of A549 cells and exhibited a time-effect relationship with miR-202 by targeted combination with IL-10 3'UTR to downregulate IL-10 expression in A549 cells.