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Objective:To investigate the effects of miRNA-126-3p (miR-126-3p) on proliferation and cycle of human monocytic leukemia THP-1 cells.Methods:Real-time fluorescence quantitative polymerase chain reaction (PCR) was used to verify the expression of miR-126-3p in THP-1 cells and monocytes isolated from the peripheral blood of healthy check-ups collected from May 2022 to January 2023 in Shanxi Cancer Hospital. The miR-126-3p overexpression vector was constructed and transfected inyo THP-1 cells (miR-126-3p overexpression group), and THP-1 cells transfected with the empty vector were used as the control. The proliferative ability of cells in each group was detected by CCK-8 assay, and the cell cycle was detected by flow cytometry. TargetScan software was applied to predict the miR-126-3p target gene as RGS3, which was verified by dual luciferase reporter gene assay. The protein expression level of RGS3 in THP-1 cells of the miR-126-3p overexpression group was detected by Western blotting.Results:The results of PCR showed that the expression level of miR-126-3p in THP-1 cells was low compared with that in monocytes isolated from peripheral blood of healthy individuals ( P < 0.05). Compared with the control group, THP-1 cells in the miR-126-3p overexpression group had low proliferative capacity after 48 h and 72 h of culture (both P < 0.05), and the cells were blocked in G 1 phase [proportion of cells in G 1 phase: (58.2±2.8)% vs. (44.1±2.4)%, P < 0.05]. The results of dual luciferase reporter gene assay showed that miR-126-3p bound to the 3'UTR of RGS3. Western blotting assay showed that the RGS3 protein expression level in THP-1 cells of the miR-126-3p overexpression group was lower than that in the control group. Conclusions:miR-126-3p may inhibit proliferation of human monocytic leukemia THP-1 cells and promote G 1-phase blockade by inhibiting the expression of RGS3.
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Objective:To investigate the expression of miRNA-522-3p (miR-522-3p) in gastric cancer tissues/cells and its regulation of RSU1 expression and to analyze the effect of miR-522-3p on biological function of gastric cancer cells in vitro.Methods:miR-522-3p relative expression levels in tumor tissues and adjacent tissues of 50 patients clinically diagnosed as gastric cancer in Shanxi Bethune Hospital from May 2019 to June 2019 were detected by using real-time quantitative polymerase chain reaction (qRT-PCR). MGC-803 cells and BGC-823 cells in gastric cancer were divided into miR-522-3p inhibitor group (transfected with miR-522-3p inhibitor) and empty vector group (transfected with empty vector). The cell proliferation was detected by using CCK-8 assay, cell scratch assay was used to detect the scratch healing ability of cells, and flow cytometry was used to detect the apoptosis. The target correlation between miR-522-3p and RSU1 was detected by using double luciferase reporter gene assay, the change of RSU1 protein was detected by using Western blot.Results:qRT-PCR showed that compared with adjacent cancer tissues, the relative expression level of miR-522-3p was up-regulated, and the difference was statistically significant (0.84±0.31 vs. 0.48±0.22, t = 2.93, P < 0.05). There were no statistically significant differences in the relative expression level of miR-522-3p in gastric cancer tissues with different tumor diameter and pathological grade (all P < 0.05). In vitro experimental assay showed that the cell proliferation rates of MGC-803 and BGC-823 cells in miR-522-3p inhibitor group at 48 h and 72 h were decreased (all P < 0.05). Furthermore, MGC-803 and BGC-823 tranfected with miR-522-3p inhibitor could effectively inhibit the scratch healing ability of MGC-803 and BGC-823 cells. Dual-luciferase reporter gene assay verified that miR-522-3p targeted to RSU1 3'UTR and affected its fluorescence activity. Western blot results showed that miR-522-3p could promote RSU1 protein expression. Conclusions:miR-522-3p is involved in the progression of gastric cancer probably via the regulation of RSU1 expression, and it may be a potential therapeutic target.
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Objective To establish the model of VX2 soft tissue xenograft in rabbit, and choose the best time for PET-CT imaging. To analyze the result of the imaging with semi-quantitative, and evaluate the feasibility of the study with VX2 soft-tissue tumor model in rabbit for PET-CT imaging. Methods 20 Japanese white rabbits were successfully implanted with VX2 tumors in thighs by the injection of tissue mass suspension.Then they were scanned with PET-CT after the diameter of the tumor is ＞2 cm. Results The diameter of the tumor＞2 cm at 14 d post implant. The average volume of tumor was (114.57±9.87) cm3.The average SUVmax of tumor's and normal tissue's in the other thigh was 5.85±0.92 and 0.22±0.09 (t=27.03, P＜0.01). SUVmax was not associated with tumor volume (r=0.02, P=0.93).Conclusion The imaging of VX2 soft tissue xenograft is clear by clinic PET-CT, and it is suitable for the study of PET-CT.PET-CT imaging performed at 14 d post implantation had the highest imaging quality.