中国病理生理杂志
中國病理生理雜誌
중국병리생리잡지
CHINESE JOURNAL OF PATHOPHYSIOLOGY
2014年
12期
2280-2283
,共4页
张洪旺%张宝刚%宋瑞卉%王汉秋%郭文君
張洪旺%張寶剛%宋瑞卉%王漢鞦%郭文君
장홍왕%장보강%송서훼%왕한추%곽문군
Raptor蛋白%胶质瘤%肿瘤侵袭%哺乳动物霉帕雷素靶蛋白%AMPK相关激酶5%基质金属蛋白酶-2%基质金属蛋白酶-9
Raptor蛋白%膠質瘤%腫瘤侵襲%哺乳動物黴帕雷素靶蛋白%AMPK相關激酶5%基質金屬蛋白酶-2%基質金屬蛋白酶-9
Raptor단백%효질류%종류침습%포유동물매파뢰소파단백%AMPK상관격매5%기질금속단백매-2%기질금속단백매-9
Raptor protein%Glioma%Neoplasm invasiveness%Mammalian target of rapamycin%AMPK-related kinase 5%Matrix metalloproteinase-2%Matrix metalloproteinase-9
目的：研究Raptor对于胶质瘤细胞侵袭能力的影响。方法：采用RNA干扰技术，向胶质瘤U87细胞转染Raptor限定性siRNA干扰质粒， Western blotting检测转染后细胞Raptor的表达水平以鉴定转染效果。体外侵袭实验检测Raptor表达降低后，U87细胞侵袭能力的变化；Western blotting检测细胞中ARK5的磷酸化情况和MMP-2、MMP-9的表达水平。免疫组织化学法检测低级别及高级别胶质瘤中Raptor的表达水平。结果：转染Rap-tor siRNA质粒的U87细胞命名为siRaptor／U87，转染对照组质粒的细胞命名为Scr／U87，转染成功的实验组细胞Raptor的表达降低。体外侵袭实验中siRaptor／U87较对照组穿透基质膜的细胞数少（ P＜0.01）。 Western blotting显示实验组细胞中磷酸化ARK5、MMP-2和MMP-9蛋白的表达水平均较对照组低。胶质瘤组织中Raptor的表达与恶化程度存在相关性（ P＜0.01）。结论：Raptor表达降低可能通过磷酸化ARK5及增加MMP-2、MMP-9的表达促进胶质瘤细胞的侵袭力。
目的：研究Raptor對于膠質瘤細胞侵襲能力的影響。方法：採用RNA榦擾技術，嚮膠質瘤U87細胞轉染Raptor限定性siRNA榦擾質粒， Western blotting檢測轉染後細胞Raptor的錶達水平以鑒定轉染效果。體外侵襲實驗檢測Raptor錶達降低後，U87細胞侵襲能力的變化；Western blotting檢測細胞中ARK5的燐痠化情況和MMP-2、MMP-9的錶達水平。免疫組織化學法檢測低級彆及高級彆膠質瘤中Raptor的錶達水平。結果：轉染Rap-tor siRNA質粒的U87細胞命名為siRaptor／U87，轉染對照組質粒的細胞命名為Scr／U87，轉染成功的實驗組細胞Raptor的錶達降低。體外侵襲實驗中siRaptor／U87較對照組穿透基質膜的細胞數少（ P＜0.01）。 Western blotting顯示實驗組細胞中燐痠化ARK5、MMP-2和MMP-9蛋白的錶達水平均較對照組低。膠質瘤組織中Raptor的錶達與噁化程度存在相關性（ P＜0.01）。結論：Raptor錶達降低可能通過燐痠化ARK5及增加MMP-2、MMP-9的錶達促進膠質瘤細胞的侵襲力。
목적：연구Raptor대우효질류세포침습능력적영향。방법：채용RNA간우기술，향효질류U87세포전염Raptor한정성siRNA간우질립， Western blotting검측전염후세포Raptor적표체수평이감정전염효과。체외침습실험검측Raptor표체강저후，U87세포침습능력적변화；Western blotting검측세포중ARK5적린산화정황화MMP-2、MMP-9적표체수평。면역조직화학법검측저급별급고급별효질류중Raptor적표체수평。결과：전염Rap-tor siRNA질립적U87세포명명위siRaptor／U87，전염대조조질립적세포명명위Scr／U87，전염성공적실험조세포Raptor적표체강저。체외침습실험중siRaptor／U87교대조조천투기질막적세포수소（ P＜0.01）。 Western blotting현시실험조세포중린산화ARK5、MMP-2화MMP-9단백적표체수평균교대조조저。효질류조직중Raptor적표체여악화정도존재상관성（ P＜0.01）。결론：Raptor표체강저가능통과린산화ARK5급증가MMP-2、MMP-9적표체촉진효질류세포적침습력。
[ ABSTRACT] AIM:To study the influence of Raptor on the invasion ability of glioma cells.METHODS: The technique of RNA interference was used.U87 cells were transfected with Raptor restricted siRNA plasmid, and the expres-sion level of Raptor in the transfected cells was detected by Western blotting.The invasive ability of the cancer cells in vitro was determined.The phosphorylation level of ARK5 and the expression of MMP-2 and MMP-9 were detected by Western blotting.The expression levels of Raptor in the tumor samples of low-grade gliomas ( WTO grade I and grade II) and high-grade gliomas (WTO grade III and grade IV) were also analyzed by immunohistochemical staining.RESULTS: Raptor siRNA was transfected into U87 cells and the cells were named siRaptor/U87 cells.The cells transfected with the control plasmid was named Scr/U87 cells.The expression level of Raptor in siRaptor/U87 cells was lower than that in Scr/U87 cells.The results of in vitro invasion assay showed that the number of siRaptor/U87 cells penetrating the Matrivgel matrix membrane was less than that of Scr/U87 cells (P<0.01).The protein expression of MMP-2 and MMP-9, and phosphoryl-ation of ARK5 protein in the cells in the experimental group were lower than those in control group.The correlation between the expression of Raptor in gliomas and the degree of deterioration was also observed ( P<0.01) .CONCLUSION: The expression of Raptor may contribute to the invasion ability of glioma cells by phosphorylation of ARK5 and increase in the levels of MMP-2 and MMP-9.
