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Objective:To investigate the mechanism of miR-1290 mediated by ultrasound microbubbles on the proliferation, apoptosis and invasion of ovarian cancer cells by regulating the expression of DKK3.Methods:Logarithmic SKOV3 cells were divided into Control group, miR-1290 NC group, microbubble treatment (MB) group, miR-1290 inhibitor group and miR-1290 inhibitor-MB group. The targeting relationship between miR-1290 and DKK3 was verified by double luciferase assay; RT-PCR was used to detect the expression of miR-1290 and DKK3 mRNA in SKOV3 cells; the activity of SKOV3 cells was detected by MTT assay; the apoptosis of SKOV3 cells was detected by flow cytometry; cell scratch test and Transwell test were used to detect the migration and invasion abilities of SKOV3 cells; Western blot was used to detect the expression of protein.Results:The double luciferase experiment showed that miR-1290 had a targeting relationship with DKK3, and miR-1290 could negatively target DKK3; Compared with miR-1290 NC group [24, 48, 72 h: (0.53 ± 0.05), (0.82 ± 0.06), (1.24 ± 0.06) ], MB group [24, 48, 72 h: (0.43 ± 0.06), (0.71 ± 0.03), (1.03 ± 0.03) ], miR-1290 inhibitor group [24, 48, 72 h: (0.41 ± 0.03), (0.66 ± 0.04), (0.78 ± 0.05) ], miR-1290 inhibitor MB group [24, 48, 72 h: (0.33 ± 0.04), (0.54 ± 0.05), (0.67 ± 0.06) ] SKOV3 cell proliferation activity decreased significantly ( P<0.05), compared with the migration rate and invasion number of SKOV3 cells in miR-1290 NC group [ (45.98 ± 4.11) %, (235.14 ± 5.78) ], the migration rate and invasion number of SKOV3 cells in MB group [ (36.77 ± 4.24) %, (189.57 ± 4.58) ], miR-1290 inhibitor group [ (32.14 ± 3.78) %, (165.35 ± 5.01) ], and miR-1290 inhibitor MB group [ (20.40 ± 3.01) %, (86.21 ± 4.23) ] decreased significantly,the expression of DKK3 mRNA and protein, apoptosis rate, apoptosis promoting protein C-Caspase-3 and Bax in SKOV3 cells increased greatly ( P<0.05) ; the above indexes of SKOV3 cells in miR-1209 NC group and Control group had no great difference ( P>0.05) . Conclusion:MiR-1290 can negatively regulate the expression of DKK3, while ultrasound microbubble can down regulate the expression of miR-1290 and up regulate the expression of DKK3, thereby inhibiting the proliferation, migration and invasion of ovarian cancer SKOV3 cells and promoting the apoptosis of cancer cells.
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OBJECTIVE:To compare the total flavonoids content and antioxidant activity in the barks,leaves,male flowers and seeds of Eucommiae ulmoides. METHODS:UV spectrophotometry was used to determine the total flavonoids content in differ-ent parts;tests was conducted to clear 2,2′-nitrilobis(3-ethylbenzothiazoline-6-sulfonic acid)(ABTS+),1,1-diphenyl-2-trinitro-phenylhydrazine(DPPH)radicals and the reducing ability of Cu2+,using half clear/reduction concentration value(IC50)as evalua-tion indexes,and vitamin C was regarded as positive control. RESULTS:The total flavonoids content of the E. ulmoides from hight to low was as follows as leaves>male flowers>barks>seeds,except there was no significant difference in barks and seeds (P>0.05),the other parts had significant differences (Pmale flowers>seeds>barks,except there was no significant difference in the indicators of leaves and male flowers(P>0.05),the other parts had significant differences(Pmale flowers>barks>seeds,there was significant difference in leaves and males flowers with barks and seeds (P<0.05). CONCLUSIONS:The content of total flavonoids in leaves and male flowers is high,and the antioxidant activity is strong,which has a great prospect of exploitation and utilization to make up for deficiencies in barks of E. ulmoides.
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OBJECTIVE@#To observe the expression of progesterone receptor (PR), interleukin-1β (IL-1β), and cyclooxygenase-2 (COX-2) induced by lipopolysaccharide (LPS) or Toll-like receptor 4 antagonist (TLR4 mAb) in decidual cells in vitro, and then to explore the effect of LPS and its antagonist on PR of decidual cells and the relation between PR and inflammatory cytokines.@*METHODS@#We isolated and cultured human decidua of early abortion in the sterile state. When the cells passaged to the 4th generation, the cells were randomly divided into 6 pore plates: A control group was added the culture medium alone; experimental group I was added 100 ng/mL of LPS; experimental group II was add 1 μg/mL of TLR4 mAb; experimental group III was added 3 μg/ mL of TLR4 mAb; experimental group IV was added 1 μg/mL of TLR4 mAb pretreatment for 24 h, and then 100 ng/mL LPS; and experimental group V was added 3 μg/mL of TLR4 mAb pretreatment for 24 h, and then 100 ng/mL LPS for 24 h culture. Subsequently, HE staining and immunofluorescence were used to observe the morphology and identify the purity of decidual cells in the 6 groups. The levels of mRNA expression of PR, IL-1β, and COX-2 were detected by reverse transcription PCR (RT-PCR).@*RESULTS@#LPS reduced the mRNA expression of PR (P<0.05), increased the mRNA expression of IL-1β and COX-2 (P<0.05). TLR4 mAb increased the mRNA expression of PR (P<0.05) and reduced the mRNA expression of IL-1β (P<0.05) after LPS-stimulated decidual cells. High concentrations of TLR4 mAb reduced the mRNA expression of COX-2 (P<0.05) after LPS stimulated decidual cells.@*CONCLUSION@#The mRNA expression of PR is reduced, and the mRNA expressions of IL-1β and COX-2 are increased after LPS-stimulated decidual cells in vitro. TLR4 mAb antagonize the role of LPS on PR, IL-1β, and COX-2.