Expression changes of miR-146a-5p and SMAD4 in thyroid follicular carcinoma tissues,their effects on proliferation,migration,and invasion of FTC-238 cells,and verification of the targeted relationship
Objective To analyze the targeted relationship between miR-146a-5p and SMAD4 by observing the ex-pression changes of microRNA-146a-5p(miR-146a-5p)and SMAD4 homolog 4(SMAD4)in follicular thyroid carcinoma(FTC)tissues and their effects on cell proliferation migration and invasion abilities,in order to explore whether miR-146a-5p promotes invasion and metastasis of FTC by targeting SMAD4.Methods We collected 30 samples of FTC tissues and 30 samples of non-tumor thyroid tissues or tissues adjacent to the tumor margin(>2 cm)from patients with FTC.We used RT-qPCR to detect miR-146a-5p and SMAD4 mRNA in the FTC tissues and adjacent tissues.The FTC-238 cells were di-vided into the miR-146a-5p mimics group and the negative control(miR-NC group),which were transfected with miR-146a-5p mimics(to overexpress miR-146a-5p in the cells)and miR-NC,respectively.CCK-8 was used to observe the cell proliferation abilities of the two groups at 12,24,and 48 h of culture.The scratch test was used to observe the migration abilities of the two groups,and the Transwell chamber experiment was used to observe the invasion abilities of the two groups.RT-qPCR and Western blotting were used to detect SMAD4 mRNA and protein in the two groups,respectively.The dual luciferase reporter gene experiment was used to verify the targeted relationship between miR-146a-5p and SMAD4.Results The relative expression level of miR-146a-5p in FTC tissues was lower than that in the adjacent tissues(P<0.05).The relative expression level of SMAD4 mRNA in FTC tissues was higher than that in adjacent tissues(P<0.05).The OD value at each time point in the miR-146a-5p mimics group was lower than that in the miR-NC group(all P<0.05).The number of migration and invasive cells after transfection in the miR-NC group was larger than that in the miR-146a mimics group(P<0.05).The relative expression levels of SMAD4 mRNA and protein in the miR-146a-5p mimics group were lower than those in the miR-NC group(both P<0.05).There was a targeted relationship between miR-146a-5p and SMAD4.Conclusion In FTC tissues,the expression of miR-146a-5p decreases while the expression of SMAD4 in-creases.Overexpression of miR-146a-5p can inhibit the proliferation,migration,and invasion of the FTC-238 cells.There is a targeted relationship between miR-146a-5p and SMAD4.MiR-146a-5p may promote the proliferation,invasion,and migration of FTC cells by targetedly inhibiting SMAD4.
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