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Objective:To study the effect of miR-122a on inhibition proliferation of laryngeal carcinoma cell line Hep-2.Meth-ods:The oligomucleotide of miR-122a was transfected into laryngeal carcinoma cell line Hep 2 cells,which were devided into three groups of A(miR-122a transfection),B(miR-122a inhibitor),C(miR-122a-NC inhibitor) and group of D(blank control).The expres-sion of miR-122a was defected by RT-PCR,and relevant protein expression was evaluated by Western blot .The cell proliferation and cell cycle were determined by MTT assy and flow cytometry ,respectively.Results:Compared to group D,miR-122a expression in Hep2 cells was obviously elevated atter miR-122a-transfected.The proliferation of Hep2 cells in group A was significantly inhibited and the cell cycle arrested at G1/G0 phase.The protein expression of CDC42 was downregulated with decreased expressions of CDK 4 and cyclin D1 in group A.Conclusion:miR-122a inhibits the proliferation activity of Hep 2 cells,suggesting that miR-122a can be taken as a po-tential candidate for gene therapy of laryngeal carcinoma .
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Aim To investigate the effects of miR-122a on blood-spinal cord barrier after spinal cord ischemia-reperfusion injury in rats.Methods Thirty-six SD rats were randomly divided into three groups:group of sham(S group),group of control(C group)and group of miR-122a antagomir(M group).Rats in S group were subjected to exposure of aorta arch but without occlusion.Spinal ischemia-reperfusion injury was induced by clamping the aorta arch for 14 min in C group and M group.Rats in M group and C group were intrathecally injected with miR-122a antagomir or antagomir control daily for three times after injury.The miR-122a expression in injured spinal cord tissue was detected by real-time PCR.The occludin expression in injured spinal cord tissue was detected by Western blot.The permeability of blood-spinal cord barrier was examined using evans blue as a vascular tracer.The neurological motor function was evaluated by Basso Beattie Bresnahan score.Results Compared with S group,the expression of miR-122a was increased,the expression of occludin was decreased,the permeability of blood-spinal cord barrier was increased,and neurological motor function score was decreased significantly in C group(P<0.05).Compared with C group,the expression of miR-122a was decreased,the expression of occludin was increased,the permeability of blood-spinal cord barrier was decreased,and neurological motor function score was increased significantly in M group(P<0.05).Conclusion miR-122a can regulate the expression of occludin and change the permeability of blood-spinal cord barrier.
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Objective To explore the role of the abnormal expression of miRNAs in the development process of non-small cell lung cancer and the feasibility of ultrasound microbubble-mediated gene therapy after transfecting antisense miRNA-224 and miRNA-122a plasmids into non-small cell lung cancer A549 cells. Methods Antisense miRNA-224 and miRNA-122a plasmids were transfected into non-small cell lung cancer A549 cells on the optimal ultrasound microbubble-mediated condition. We set up a control group. The cell proliferation activity, apoptosis, invasion ability were detected by MTT assay, Annexin V-PE, Transwell invasion experiment and colony formation assay, respectively. Results The expression of miRNA-224 decreased and the expression of miRNA-122a rose after the plasmids of target genes were transfected into non-small cell lung cancer A549 cells, and there were significant differences when compared with those of the control group (P < 0.05). After the plasmids of target genes were transfected into A549 cells, the growth of antisense miRNA-224 and miRNA-122a were inhibited, and the differences were significant as compared with the control group (P < 0.05). Besides, the inhibition of miRNA-122a group was the most significant and there was statistically significant difference as compared with miRNA-224 group (t = −4.694, P = 0.009). After the plasmids of target genes were transfected into A549 cells, the proportion of apoptotic cells increased, the invasive cells were decreased and the clone ability reduced, and also there was a significant difference as compared with those of the control group (P < 0.05). What's more, the apoptotic peak appeared in miRNA-122a group. Its invasion ability decreased most obviously (40.25 ± 3.97/visual field), the number of clone ability was 104.93 ± 4.87 and the inhibitory effect was the most obviously. There was statistically significant difference as compared with other groups (P < 0.05). Conclusions A549 cells transfected by ultrasound microbubble-mediated antisense miRNA-224 and miRNA-122a plasmids possessed good transfection efficiency. The cell growth, invasion and colony-forming abilities of transfected A549 cells were suppressed, which laid a solid foundation for the gene therapy of non-small cell lung cancer.
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OBJECTIVE@#To explore the role of the abnormal expression of miRNAs in the development process of non-small cell lung cancer and the feasibility of ultrasound microbubble-mediated gene therapy after transfecting antisense miRNA-224 and miRNA-122a plasmids into non-small cell lung cancer A549 cells.@*METHODS@#Antisense miRNA-224 and miRNA-122a plasmids were transfected into non-small cell lung cancer A549 cells on the optimal ultrasound microbubble-mediated condition. We set up a control group. The cell proliferation activity, apoptosis, invasion ability were detected by MTT assay, Annexin V-PE, Transwell invasion experiment and colony formation assay, respectively.@*RESULTS@#The expression of miRNA-224 decreased and the expression of miRNA-122a rose after the plasmids of target genes were transfected into non-small cell lung cancer A549 cells, and there were significant differences when compared with those of the control group (P < 0.05). After the plasmids of target genes were transfected into A549 cells, the growth of antisense miRNA-224 and miRNA-122a were inhibited, and the differences were significant as compared with the control group (P < 0.05). Besides, the inhibition of miRNA-122a group was the most significant and there was statistically significant difference as compared with miRNA-224 group (t = -4.694, P = 0.009). After the plasmids of target genes were transfected into A549 cells, the proportion of apoptotic cells increased, the invasive cells were decreased and the clone ability reduced, and also there was a significant difference as compared with those of the control group (P < 0.05). What's more, the apoptotic peak appeared in miRNA-122a group. Its invasion ability decreased most obviously (40.25 ± 3.97/visual field), the number of clone ability was 104.93 ± 4.87 and the inhibitory effect was the most obviously. There was statistically significant difference as compared with other groups (P < 0.05).@*CONCLUSIONS@#A549 cells transfected by ultrasound microbubble-mediated antisense miRNA-224 and miRNA-122a plasmids possessed good transfection efficiency. The cell growth, invasion and colony-forming abilities of transfected A549 cells were suppressed, which laid a solid foundation for the gene therapy of non-small cell lung cancer.