Study on mechanism of miR-19b-3p targeting IGF-1 in the development of choriocarcinoma
Objective To explore the mechanism of miR-19b-3p targeting insulin like growth factor-1(IGF-1)in the development of choriocarcinoma.Methods Human chorionic cancer cell line JEG-3 was selected as the research object and divided into five groups:group A:miR-19b-3p mimics group;group B:miR-19b-3p mimics NC group;group C:miR-19b-3p inhibitor group;group D:Mir-19b-3p inhibitor group;group E:blank control group.Groups A,B,C,and D were transfected accordingly to the grouping,and group E did not receive any treatment.The expression levels of miR-19b-3p RNA and IGF-1 protein in five groups were detected by Real-time PCR and Western blot,respectively,and cell proliferation was measured by CCK-8 method.Scratch assay and Transwell cell invasion assay were used to determine cell migration and invasion ability in each group,and then compared.Dual-luciferase reporter assay was used to verify the targeting relationship between miR-19b-3p and IGF-1.Results Compared with(0.98±0.13)in group E,miR-19b-3p mRNA expression level of(2.23±0.16)was significantly up-regulated in group A,and miR-19b-3p mRNA expression level of(0.73±0.08)was significantly down-regulated in group C(P<0.05).Compared with(1.03±0.03)nmol/L in group E,IGF-1 protein expression level of(1.48±0.12)nmol/L in group A was significantly up-regulated,and that of(0.76±0.05)nmol/L in group C was significantly down-regulated(P<0.05).At 72 h,compared with(5.03±0.14)%in group E,the cell proliferation viability of(10.34±0.87)%in group A was significantly increased,and the cell proliferation viability of(2.42±0.13)%in group C was significantly down-regulated(P<0.05).Compared with(1.09±0.07)mm in group E,the cell scratch migration distance was significantly increased in group A[(1.45±0.08)mm]and significantly decreased in group C[(0.73±0.07)mm](P<0.05).The number of invaded cells was significantly increased in group A[(173.00±4.00)]and significantly decreased in group C[(82.00±1.00)]compared to(103.00±7.00)in group E(P<0.05).The results of dual luciferase assay showed that the cellular luciferase activity of miR-19b-3p transfected with WT-IGF-1was(0.57±0.08)Kat,which was lower than(0.95±0.06)Kat of miR-NC(P<0.05).The cellular luciferase activity of miR-19b-3p transfected MUT-IGF-1 was(0.98±0.05)Kat,which was not significantly different from(0.96±0.06)Kat of miR-NC(P>0.05).Conclusion Upregulation of IGF-1 by miR-19b-3p targeting is involved in promoting proliferation,invasion and migration of JEG-3 cells in choriocarcinoma.
ChoriocarcinomamiR-19b-3pInsulin like growth factor-1ProliferateInvasion
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