CAJ | 학술논문

目的：观察沉默钙离子相关蛋白（S100A4）表达对鼻咽癌细胞株CNE2凋亡及侵袭的影响。方法培养CNE2细胞，分为空白组、对照组及实验组。实验组细胞通过Lipofectamine2000转染S100A4 siRNA，对照组细胞通过Lipofectamine2000转染空质粒，空白组细胞未做转染。 Western blotting 法检测CNE2细胞中的S100A4蛋白。real-time PCR法检测S100 A4 mRNA。流式细胞术检测细胞凋亡情况。 Transwell实验检测细胞侵袭能力。结果实验组、空白组、对照组S100A4 mRNA相对表达量分别为0．6004±0．05、1．0000±0．00、0．8941±0．09，S100A4蛋白相对表达量分别为0．22±0．016、0．42±0．022、0．39±0．022。实验组细胞中S100A4 mRNA、蛋白表达量与空白组及对照组相比，P均＜0．05。实验组、空白组、对照组细胞凋亡率分别为45．87％、3．49％、2．49％，实验组与空白组及对照组相比，P均＜0．05。 Transwell实验结果示实验组穿膜细胞数为（206±22）个，空白组和对照组分别为（329±12）、（347±21）个。实验组穿膜细胞数与空白组、对照组相比，P均＜0．05。结论沉默S100A4基因表达后，CNE2细胞凋亡增多，细胞侵袭能力渐弱。
목적：관찰침묵개리자상관단백（S100A4）표체대비인암세포주CNE2조망급침습적영향。방법배양CNE2세포，분위공백조、대조조급실험조。실험조세포통과Lipofectamine2000전염S100A4 siRNA，대조조세포통과Lipofectamine2000전염공질립，공백조세포미주전염。 Western blotting 법검측CNE2세포중적S100A4단백。real-time PCR법검측S100 A4 mRNA。류식세포술검측세포조망정황。 Transwell실험검측세포침습능력。결과실험조、공백조、대조조S100A4 mRNA상대표체량분별위0．6004±0．05、1．0000±0．00、0．8941±0．09，S100A4단백상대표체량분별위0．22±0．016、0．42±0．022、0．39±0．022。실험조세포중S100A4 mRNA、단백표체량여공백조급대조조상비，P균＜0．05。실험조、공백조、대조조세포조망솔분별위45．87％、3．49％、2．49％，실험조여공백조급대조조상비，P균＜0．05。 Transwell실험결과시실험조천막세포수위（206±22）개，공백조화대조조분별위（329±12）、（347±21）개。실험조천막세포수여공백조、대조조상비，P균＜0．05。결론침묵S100A4기인표체후，CNE2세포조망증다，세포침습능력점약。
Objective To observe the effect of silencing calcium ion associated protein ( S100 A4 ) expression on ap-optosis and invasion of nasopharyngeal carcinoma cell line CNE 2.Methods The CNE2 cells were cultured , and then were divided into the blank group , control group and experimental group .The cells of the experimental group were trans-fected with S100A4siRNA by Lipofectamine2000, the control group was transfected with CON (empty plasmid) by Lipo-fectamine2000 , and the cells in the blank group did not receive special transfection .Western blotting was used to detect the S100A4 protein in CNE2 cells, and the expression of S100A4 mRNA was detected by real-time PCR.The apoptosis was detected by flow cytometry .Transwell assay was used to detect cell invasion .Results The relative expression levels of S100A4 mRNA in the experimental group, blank control group and control group were respectively 0.600 4 ±0.05, 1.000 0 ±0.00 and 0.894 1 ±0.09, and the relative expression levels of S100A4 protein were respectively 0.22 ±0.016, 0.42 ±0.022 and 0.39 ±0.022.The expression of S100A4 mRNA and protein in the experimental group was lower than that in the control group and the blank control group (all P<0.05).The apoptosis rates of the experimental group , blank group and control group were 45.87%, 3.49%and 2.49%.The apoptosis rates in the experimental group and the blank group were lower than that in the control group (all P<0.05).Transwell experimental results show that the number of cells passing through the cell membrane was (206 ±22), the blank group and the control group were (329 ±12) and (347 ±21).Significant difference was found in the number of cells between the experimental group and the blank group , the control group (all P<0.05).Conclusion After silencing S100A4 gene expression, the apoptosis of CNE2 cells was in-creased, and cell invasion ability was decreased .