Study On The Mechanism Of MiR-152-3p Regulating Papillary Thyroid Cancer Via TGFA

Objective: The purpose of this study is to make sure whether miR-152-3p plays a key role in the growth of papillary thyroid carcinoma(PTC)cells and to explore its targeting relationship.Methods: The expression level of miR-152-3p was detected by real-time fluorescence quantitative polymerase chain reaction.The object of study was 20 pairs of PTC tissue and paracancerous thyroid tissue.MiR-152-3p mimics and mi R-152-3p inhibitor were transfected into PTC cell line(KTC-1),and negative control group was set up.PCR was used to test the transfection efficiency of each group of miR-152-3p.Through cell counting Kit-8,the proliferation ability of PTC cell line can be detected,and the cell migration ability can be reflected by the ratio of scratch width in cell scratch experiment,and the invasion ability of KTC-1 in each group can be tested by Transwell cell chamber method.TGFA was screened as the target of miR-152-3p via targeted prediction software.Double luciferase reporter gene was used to detect the complementary combination of TGFA and miR-152-3p.The expression of TGFA from protein level and mRNA level in cell lines was tested respectively by Western blot and PCR,which we could analyze effect of miR-152-3p on TGFA.The expression of TGFA in PTC tissue was detected by PCR.After that,TGFA overexpression plasmid and TGFA siRNA were transfected to KTC-1,and the corresponding negative control group was also set,at the same time,transfection efficiency should also be measured.CCK-8 kit and Transwell chamber method wereused to detect the proliferation and invasion of cancer cells in TGFA group.Results: The expression of miR-152-3p in PTC tissues and PTC cells was lower than that in parathyroid tissues.After the transfection experiment,PCR showed that the transfection efficiency was effective.In vitro,the proliferation,migration and invasion of KTC-1 cells wereobviously enhanced in mi R-152-3p-inhibitor group compared with the negative control group and miR-152-3p-mimics group.The potential target gene TGFA of miR-152-3p was determined by using 3 publicly available miRNA target prediction tools.This targeting relationship was confirmed by the detection of double luciferase reporter gene,and the results of RT-PCR and WB showed that mi R-152-3p inhibited the expression of TGFA in one way.In papillary thyroid carcinoma,the real-time fluorescence quantitative polymerase chain reaction showed that the average relative quantity(RQ)of TGFA was higher than that of the adjacent tissues.The follow-up experiments further proved that the proliferation and invasion ability of cancer cells in the TGFA(OE)transfection group was stronger than that in the NC group and TGFA(KD)transfection group.TGFA was regarded as a positive regulator of PTC cell proliferation and invasion.Conclusion: Our study found that the expression level of miR-152-3p in papillary thyroid carcinoma tissues is lower than that in the adjacent tissues,and TGFA is an important link in the PTC development and even metastasis.What's more,at the level of gene transcription and protein expression,miR-152-3p can inhibit the expression of TGFA,so as to inhibit the proliferation,migration andinvasion of KTC-1 cell line.These results may be potential targets for the study of the pathogenesis and development of PTC,and even for the prevention and treatment of PTC.