CAJ | 학술논문

目的：将 Shh 基因用慢病毒载体转染甲状腺乳头状癌（PTC）原代细胞中，观察 Shh 对 PTC 细胞体外生物学性状的影响。方法在建立慢病毒载体 pGC-FU-Shh-GFP 转染 PTC 细胞，稳定过表达 Shh 蛋白模型基础上，设立转染实验组和阴性对照组。MTT 法检测细胞增殖能力，流式细胞术检测细胞周期，Transwell 实验检测细胞迁移能力，检测 Sonic hedgehog 通路相关基因 mRNA 与蛋白表达变化，与阴性对照组检测结果进行比较。结果高表达Shh 细胞生长曲线较陡直，生长速度较阴性对照组显著提高（P ＜0．05）；Shh-PTC 细胞24 h 后 S 期上升至20．88％， Shh-PTC 细胞48 h 后 G1期上升至83．78％；实验组穿透滤过膜细胞数量明显增加（P ＜0．05）。转染后 SMO、Gli1、PTCH 蛋白的表达均高于对照组，表达量分别是对照组的1．154倍，1．062倍和1．205倍。结论Shh 明显促进 PTC细胞的增殖、迁移和侵袭能力。在 PTC 的发生、发展中 Sonic hedgehog 通路激活是激发或促进因素，该信号通路可能促进 PTC 的生长、转移。
목적：장 Shh 기인용만병독재체전염갑상선유두상암（PTC）원대세포중，관찰 Shh 대 PTC 세포체외생물학성상적영향。방법재건립만병독재체 pGC-FU-Shh-GFP 전염 PTC 세포，은정과표체 Shh 단백모형기출상，설립전염실험조화음성대조조。MTT 법검측세포증식능력，류식세포술검측세포주기，Transwell 실험검측세포천이능력，검측 Sonic hedgehog 통로상관기인 mRNA 여단백표체변화，여음성대조조검측결과진행비교。결과고표체Shh 세포생장곡선교두직，생장속도교음성대조조현저제고（P ＜0．05）；Shh-PTC 세포24 h 후 S 기상승지20．88％， Shh-PTC 세포48 h 후 G1기상승지83．78％；실험조천투려과막세포수량명현증가（P ＜0．05）。전염후 SMO、Gli1、PTCH 단백적표체균고우대조조，표체량분별시대조조적1．154배，1．062배화1．205배。결론Shh 명현촉진 PTC세포적증식、천이화침습능력。재 PTC 적발생、발전중 Sonic hedgehog 통로격활시격발혹촉진인소，해신호통로가능촉진 PTC 적생장、전이。
Objective To investigate biological behaviors’changes of papillary thyroid carcinoma cell's biological properties by constructing stable Shh-over-expressing cell line and aim to clarify the effects and possible Shh signal transduction mechanism.Methods PTC cells were infected with lentiviruse pGC-FU-Shh-GFP.Shh protein expression was detected.The proliferative and migratory activities of every group were detected using cell active assay(MTT)and Transwell cell migratory assay.Flow Cytometry (FCM)was performed to observe the effects of Shh over-expression on the distribution of cell cycle of PTC cells.Detecting the expressions of Shh signaling pathway related genes (SMO, Gli1,PTCH)of PTC-Shh cells.Results Overexpression of Shh could significantly improve proliferation of PTC cells (P <0.05)as compared with the blank control group.Overexpression of Shh caused the increase of the percentage of G1 stage of PTC cell(83.78%).Transwell cell migratory assay suggests that improvement of Shh expression suppress migratory activity of PTC cell (P <0.05).Comparing with PTC-vector cells,the expressions of genes SMO,Gli1 and PTCH increased in PTC-Shh cells.Conclusion Shh could promote the biological behaviors of PTC cell.To activate Shh signaling pathway is the promoting factor and the occurrence in PTC.The report suggests that the mechanism may stimulate the growth and metastasis of the PTC.