重庆医学
重慶醫學
중경의학
CHONGQING MEDICAL JOURNAL
2015年
1期
7-9,13
,共4页
龙新华%刘志礼%刘家明%陈宣银%王恒%周扬%张志宏
龍新華%劉誌禮%劉傢明%陳宣銀%王恆%週颺%張誌宏
룡신화%류지례%류가명%진선은%왕항%주양%장지굉
骨肉瘤%微小RNAs%转染%脂肪酸合成酶%靶向调控
骨肉瘤%微小RNAs%轉染%脂肪痠閤成酶%靶嚮調控
골육류%미소RNAs%전염%지방산합성매%파향조공
osteosarcoma%MicroRNAs%transfection%fatty acid synthase%targeting regulation
目的：检测可能调控脂肪酸合成酶（FASN）基因表达的miRNAs分子在不同侵袭力骨肉瘤细胞中的表达，筛选出靶向调控FASN基因的miRNAs分子，为研究骨肉瘤转移调控机制奠定基础。方法应用miRNA和microrna．org在线软件，预测可能靶向调控FASN基因（NM＿004104）表达的miRNAs分子；实时定量PCR（RT‐PCR）检测所预测到的miRNAs在骨肉瘤细胞HOS和U2‐OS中的表达；Westernblot检测HOS和U2‐OS细胞中FASN蛋白表达；Transwell侵袭实验检测HOS和U2‐OS细胞侵袭力。结果2种生物信息学预测方法预测结果均显示，FASN基因可能是miR‐195／15a／15b／16／424／497分子的靶基因；RT‐PCR结果显示，miR‐424在HOS细胞中表达水平显著高于在U2‐OS细胞中的表达水平；FASN在U2‐OS细胞中表达水平显著高于HOS细胞中的表达水平；U2‐OS细胞侵袭能力明显高于HOS细胞的侵袭力。结论miR‐424在骨肉瘤细胞中表达水平与细胞侵袭力呈负相关，miR‐424很可能通过靶向调控FASN基因影响骨肉瘤细胞侵袭转移。
目的：檢測可能調控脂肪痠閤成酶（FASN）基因錶達的miRNAs分子在不同侵襲力骨肉瘤細胞中的錶達，篩選齣靶嚮調控FASN基因的miRNAs分子，為研究骨肉瘤轉移調控機製奠定基礎。方法應用miRNA和microrna．org在線軟件，預測可能靶嚮調控FASN基因（NM＿004104）錶達的miRNAs分子；實時定量PCR（RT‐PCR）檢測所預測到的miRNAs在骨肉瘤細胞HOS和U2‐OS中的錶達；Westernblot檢測HOS和U2‐OS細胞中FASN蛋白錶達；Transwell侵襲實驗檢測HOS和U2‐OS細胞侵襲力。結果2種生物信息學預測方法預測結果均顯示，FASN基因可能是miR‐195／15a／15b／16／424／497分子的靶基因；RT‐PCR結果顯示，miR‐424在HOS細胞中錶達水平顯著高于在U2‐OS細胞中的錶達水平；FASN在U2‐OS細胞中錶達水平顯著高于HOS細胞中的錶達水平；U2‐OS細胞侵襲能力明顯高于HOS細胞的侵襲力。結論miR‐424在骨肉瘤細胞中錶達水平與細胞侵襲力呈負相關，miR‐424很可能通過靶嚮調控FASN基因影響骨肉瘤細胞侵襲轉移。
목적：검측가능조공지방산합성매（FASN）기인표체적miRNAs분자재불동침습력골육류세포중적표체，사선출파향조공FASN기인적miRNAs분자，위연구골육류전이조공궤제전정기출。방법응용miRNA화microrna．org재선연건，예측가능파향조공FASN기인（NM＿004104）표체적miRNAs분자；실시정량PCR（RT‐PCR）검측소예측도적miRNAs재골육류세포HOS화U2‐OS중적표체；Westernblot검측HOS화U2‐OS세포중FASN단백표체；Transwell침습실험검측HOS화U2‐OS세포침습력。결과2충생물신식학예측방법예측결과균현시，FASN기인가능시miR‐195／15a／15b／16／424／497분자적파기인；RT‐PCR결과현시，miR‐424재HOS세포중표체수평현저고우재U2‐OS세포중적표체수평；FASN재U2‐OS세포중표체수평현저고우HOS세포중적표체수평；U2‐OS세포침습능력명현고우HOS세포적침습력。결론miR‐424재골육류세포중표체수평여세포침습력정부상관，miR‐424흔가능통과파향조공FASN기인영향골육류세포침습전이。
Objective To detect the exppresion level of miRNAs which may target fatty acid synthase(FASN) in osteosarcoma (OS) cells of diverse invasion ability ,and to select the miRNAs target regulating FASN ,acting as the basis to investigate the mech‐anism of OS metastasis .Methods The miRNA and microrna .org online software were adopted to forecast the miRNAs that might target regulate FASN(NM_004104);RT‐PCR was used to detect the expression level of the predicted miRNAs in HOS and U2‐OS cells ;the protein expression of FASN was detected by Western blot ;Transwell invasion assay was used to evaluated the invasive a‐bility of HOS and U2‐OS cells .Results Two prediction methods all showed that miR‐195/15a/15b/16/424/497 molecular might be target FASN gene;RT‐PCR result showed that the expression level of miR‐424 in HOS cells was significantly higher than that in U2‐OS cells ;the expression level of FASN was significantly higher in U2‐OS cells than that in HOS cells ;the invasive ability of U2‐OS cells is significantly higher than HOS cells .Conclusion The expression level of miR‐424 in OS cells may be negatively related to the invasive ability ,and miR‐424 likely may affect OS metastasis via targeting FASN .
