中华口腔医学研究杂志（电子版）
中華口腔醫學研究雜誌（電子版）
중화구강의학연구잡지（전자판）
CHINESE JOURNAL OF STOMATOLOGICAL RESEARCH(ELECTRONIC VERSION)
2014年
6期
454-458
,共5页
焦国华%阚智慧%高洪丽%张志光
焦國華%闞智慧%高洪麗%張誌光
초국화%감지혜%고홍려%장지광
舌肿瘤%肿瘤,鳞状细胞%RNA干扰%营养缺乏自噬因子1
舌腫瘤%腫瘤,鱗狀細胞%RNA榦擾%營養缺乏自噬因子1
설종류%종류,린상세포%RNA간우%영양결핍자서인자1
Tongue neoplasms%Neoplasms,squamous cell%RNA interference%Nutrient-deprivation autophagy factor-1deprivation autophagy factor-1
目的：使用小干扰RNA(siRNA)干扰舌鳞状细胞癌Tca8113细胞中营养缺乏自噬因子1(NAF1)的表达,观察对NAF1基因的沉默效果,并测定沉默后Tca8113细胞生长增殖侵袭能力的变化。方法建立小干扰RNA(si-NAF1)组、阴性对照(si-NC)组、对照(vector)组。采用反转录聚合酶链反应(RT-PCR)法和Western blotting法检测NAF1、细胞周期素D1(cyclin D1)、基质金属蛋白酶2(MMP-2)的表达。 MTT实验检测细胞增殖能力,平板克隆形成实验检测细胞克隆形成能力,Transwell法观察各组细胞侵袭能力变化。结果与vector组相比,NAF1-siRNA干预Tca8113细胞后,si-NAF1组NAF1的mRNA和蛋白表达水平都明显降低(x2=25.65,t=-17.1,P<0.05),cyclin D1、MMP-2蛋白表达水平降低(tcyclin D1=-14.7,tMMP-2=-9.6,P<0.05),细胞增殖能力明显降低(t=-36.77,P<0.05),克隆形成能力明显减弱(t=-12.33,P<0.05),侵袭能力明显降低,si-NAF1组穿过Transwell膜的细胞少于vector组。结论通过siRNA技术降低NAF1表达可能通过降低cyclin D1水平抑制舌鳞状细胞癌Tca8113细胞的增殖能力,通过影响MMP-2蛋白表达水平,抑制Tca8113细胞的侵袭能力。
目的：使用小榦擾RNA(siRNA)榦擾舌鱗狀細胞癌Tca8113細胞中營養缺乏自噬因子1(NAF1)的錶達,觀察對NAF1基因的沉默效果,併測定沉默後Tca8113細胞生長增殖侵襲能力的變化。方法建立小榦擾RNA(si-NAF1)組、陰性對照(si-NC)組、對照(vector)組。採用反轉錄聚閤酶鏈反應(RT-PCR)法和Western blotting法檢測NAF1、細胞週期素D1(cyclin D1)、基質金屬蛋白酶2(MMP-2)的錶達。 MTT實驗檢測細胞增殖能力,平闆剋隆形成實驗檢測細胞剋隆形成能力,Transwell法觀察各組細胞侵襲能力變化。結果與vector組相比,NAF1-siRNA榦預Tca8113細胞後,si-NAF1組NAF1的mRNA和蛋白錶達水平都明顯降低(x2=25.65,t=-17.1,P<0.05),cyclin D1、MMP-2蛋白錶達水平降低(tcyclin D1=-14.7,tMMP-2=-9.6,P<0.05),細胞增殖能力明顯降低(t=-36.77,P<0.05),剋隆形成能力明顯減弱(t=-12.33,P<0.05),侵襲能力明顯降低,si-NAF1組穿過Transwell膜的細胞少于vector組。結論通過siRNA技術降低NAF1錶達可能通過降低cyclin D1水平抑製舌鱗狀細胞癌Tca8113細胞的增殖能力,通過影響MMP-2蛋白錶達水平,抑製Tca8113細胞的侵襲能力。
목적：사용소간우RNA(siRNA)간우설린상세포암Tca8113세포중영양결핍자서인자1(NAF1)적표체,관찰대NAF1기인적침묵효과,병측정침묵후Tca8113세포생장증식침습능력적변화。방법건립소간우RNA(si-NAF1)조、음성대조(si-NC)조、대조(vector)조。채용반전록취합매련반응(RT-PCR)법화Western blotting법검측NAF1、세포주기소D1(cyclin D1)、기질금속단백매2(MMP-2)적표체。 MTT실험검측세포증식능력,평판극륭형성실험검측세포극륭형성능력,Transwell법관찰각조세포침습능력변화。결과여vector조상비,NAF1-siRNA간예Tca8113세포후,si-NAF1조NAF1적mRNA화단백표체수평도명현강저(x2=25.65,t=-17.1,P<0.05),cyclin D1、MMP-2단백표체수평강저(tcyclin D1=-14.7,tMMP-2=-9.6,P<0.05),세포증식능력명현강저(t=-36.77,P<0.05),극륭형성능력명현감약(t=-12.33,P<0.05),침습능력명현강저,si-NAF1조천과Transwell막적세포소우vector조。결론통과siRNA기술강저NAF1표체가능통과강저cyclin D1수평억제설린상세포암Tca8113세포적증식능력,통과영향MMP-2단백표체수평,억제Tca8113세포적침습능력。
Objective To investigate the inhibitory effect of small interfering RNA (siRNA) on the expression of nutrient-deprivation autophagy factor-1 (NAF1) in human tongue cancer Tca8113 cell line via transfected with the special siRNA. The biological behaviors, including cell proliferation and metastasis of the transduced cells were investigated as well. Methods The cultured Tca8113 cells were divided into three groups including si-NAF1 group, si-NC group and the vector group. MTT assay and plate cloning formation assay were used to identify the effect of NAF1-siRNA on the cell proliferation. Transwell assay.was used to investigate the invasion capability of the Tca8113 cells. The RNA and protein levels of functional biomarkers such as cyclin D1 and MMP-2 were detected through RT-PCR and Western blot assay. Results NAF1 expression in Tca8113 cells transfected with NAF1-siRNA was significantly down regulated at both the RNA and protein levels compared with those in normal Tca8113 cells (x2=25.65,t=-17.1,P<0.05), accompany with the decreases of cyclin D1 and MMP-2 protein(tcyclin D1=-14.7,tMMP-2=-9.6,P<0.05). The proliferation and cloning formation of Tca8113 cells were obviously inhibited in si-NAF1 group compared with the vector group (t=-36.77,t=-12.33,P<0.05). The cell invasion of Tca8113 cells was obviously inhibited in si-NAF1 group compared with the vector group. Conclusion Down regulation of NAF1 inhibited tongue cancer cells proliferation and metastasis, which might mediated via cyclin D1 and MMP-2 protein.
