A Preliminary Study Of Meis2 Involved In The Role Of TGF-β/Smad Signaling Pathway In Non-small Cell Lung Cancer

Objective(1)To investigate whether there is correlation between Meis2,Smad4 and Smad7 protein expression in non-small cell lung cancer(NSCLC)cells;(2)Initially explore the mechanism of Meis2 participating in TGF-β / Smad signaling pathway in NSCLC cells.Methods(1)Collect 80 cases of paraffin-embedded NSCLC and benign disease tissue specimens resected and surgically diagnosed in the First People’s Hospital of Yichang City from February 2012 to February 2019,except for malignant tumors and metastatic tumors in other parts.Immunohistochemical method was used to detect the protein expression level of Meis2,Smad4 and Smad7 in pathological tissues;(2)PLKO.1-Meis2-shRNA was infected into A549 cells by lentivirus transfection,and real-time quantitative fluorescent polymerase chain reaction was used(RT-qPCR)and Western-Blot(WB)methods to detect gene suppression efficiency;(3)TGF-β / Smad signaling pathway related factor mRNA expression levels in A549 cells after RT-qPCR infection;(4)WB detection in A549 cells TGF-β / Smad signaling pathway related protein expression levels.Results(1)The positive expression rate of Meis2 protein in lung cancer tissues is higher than that of benign lesions,and the expression of Smad4 protein in malignant tumor tissues is lower than that of benign lesions;Smad7 protein expression in malignant tumor tissues is significantly higher than that of benign lesions;Positive correlation;(2)The plasmid p LKO.1-Meis2 was successfully transfect into 293 FT cells,the virus solution was collected to infect A549 cells,RT-qPCR was used to detect the decrease of Meis2 mRNA level,Western-Blot method was used to detect the decrease of Meis2 gene protein expression,suggesting Transfection was successful;(3)RT-qPCR detection showed that compared with the control group,the mRNA expression of Smad2,Smad3,and Smad7 were suppressed,while the mRNA expression of TGF-β1,TGF-β2,and Smad4 was enhanced;(4)Western-Blot method showed that compared with the control group(Meis2-shRNA-GFP group),the expression of Smad4 protein was increased 1.4-1.8 times,and the protein expression of Smad2,Smad3,and Smad7 decreased.Conclusion(1)Meis2 gene is involved in the development of non-small cell lung cancer;(2)Meis2 gene participates in TGF-β-mediated Smad signaling pathway in NSCLC cells.