RESUMO
Objective To investigate the effects and molecular mechanism of circular RNA (circ) _0003028 on proliferation, migration and invasion of human liver cancer cells. Methods Liver cancer cell line Huh7 were divided into small interfering RNA(si)-NC group, si-circ_0003028 group, microRNA(miR) -NC group, miR-498 mimics group, si-circ _0003028+anti-miR-NC group and si-circ_0003028+anti-miR-498 group. Real-time PCR was used to detect the expression levels of circ _ 0003028 and miR-498 in liver cancer tissues and cells of each groups. MTT was used to detect cell proliferation. Transwell was used to detect cell migration and invasion. Western blotting was used to detect protein expression. Dual luciferase reporter experiment was used to detect the target regulation relationship between circ_0003028 and miR-498. Results The expression level of circ_0003028(0.98±0.02 vs 1. 36±0. 01) increased and the expression level of miR-498(0. 98±0. 02 vs 0. 63±0. 02) decreased in liver cancer tissues (P<0. 05). After inhibiting the expression of circ_0003028 or overexpression of miR-498- the expression levels of Ki-67(0. 85±0. 02 vs 0. 41±0. 02 or 0. 95±0. 11 vs 0. 37±0. 02)- matrix metalloprotein(MMP)-2(0. 71±0. 02 vs 0. 43±0. 03 or 0. 83±0. 02 vs 0. 41±0. 03)- and MMP-9 (0. 74±0. 02 vs 0. 37±0. 02 or 0. 78±0. 02 vs 0. 39±0. 02) proteins in Huh7 cells decreased- and cell viability(1. 53± 0. 03 vs 1. 05±0. 02 or 1. 68±0. 02 vs 1. 11±0. 02) decreased- the number of migration(111. 40±2. 12 vs 77. 22±2. 38 or 108. 90±2. 30 vs 78. 44 ± 1. 46) and invasion ( 87. 89 ± 2. 18 vs 49. 78 ± 1. 98 or 80. 22 ± 1. 79 vs 38. 22 ± 1. 52) cells decreased- and the protein expression levels of suppressor with morphogenesis effect on genitalia-1( SMG-1) (0. 76±0. 02 vs 1. 39±0. 02 or 0. 79±0. 02 vs 1. 39±0. 02)- p53(0. 77±0. 02 vs 1. 24±0. 03 or 0. 82±0. 03 vs 1. 45±0. 03)- and p53- ser15(0. 78±0. 03 vs 1. 50±0. 02 or 0. 82±0. 02 vs 1. 49±0. 04) increased (P<0. 05). circ_0003028 targeted regulation of miR-498- and silencing miR-498 reversed the effects of inhibiting the expression of circ_0003028 on the proliferation- migration and invasion of Huh7 cells. Conclusion Inhibiting the expression of circ_0003028 can inhibit the proliferation- migration and invasion of liver cancer cells by targeting miR-498 to affect the SMG-1/ p53 signaling pathway.
RESUMO
Objective To observe the effect of electroacupuncture (EA) at Baihui (GV20), Dazhui (GV14) and Shenshu (BL23) acu-points on cognitive function and the synapse of neurons in hippocampal CA1 in SAMP8 mice,to explore the mechanism of EA in the treat-ment of Alzheimer's disease(AD).Methods A total of 24 seven-month-old SAMP8 mice were randomly divided into model group(n=12) and EA group (n=12), and the same age SAMR1 mice were as control group (n=12).The EA group accepted EA at Baihui, Dazhui and Shenshu for 30 days.They were assessed with Morris maze test.The expression of synaptophysin(SYN)and postsynaptic density protein 95(PSD95)in hippocampal CA1 region were detected with immunohistochemistry.The morphology and density of synapse in hippocampal CA1 region was observed with transmission electron microscopy.Results Compared with the model group,the latency of Morris maze de-creased in EA group(P<0.05),the time staying in the quadrant of the platform increased(P<0.05),as well as the number passing the origi-nal platform(P<0.05),with the more expression of SYN and PSD95 in hippocampal CA1 region(P<0.001),and more and completed syn-apse.Conclusion EA can improve the learning and memory ability of SAMP8 mice by increasing the expression of SYN and PSD95 to pro-tect the ultrastructure of synapses in hippocampal CA1 region.