中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2015年
4期
770-774
,共5页
孙树鹏%王琼%王修玉%尚超%王金环
孫樹鵬%王瓊%王脩玉%尚超%王金環
손수붕%왕경%왕수옥%상초%왕금배
微丝相关蛋白1L2%胶质瘤%侵袭%迁移
微絲相關蛋白1L2%膠質瘤%侵襲%遷移
미사상관단백1L2%효질류%침습%천이
Actin filament associated protein 1-like 2%Glioma%Invasion%Migration
目的 观察下调胶质瘤细胞株LN229中微丝相关蛋白1L2 (AFAP1L2)的表达对细胞侵袭和迁移能力的影响.方法 将LN229细胞分为空白对照组、无义序列小于扰RNA (siRNA)转染组和AFAP1 L2 siRNA转染组进行细胞培养,细胞转染siRNA后,采用Transwell、划痕实验检测细胞侵袭和迁移能力;采用Western blot检测AFAP1L2、基质金属蛋白酶(MMP)-2、MMP-9的表达水平.结果 转染siRNA后,Western blot检测结果显示:空白对照组、无义序列siRNA转染组、AFAP1L2 siRNA转染组AFAP1L2蛋白相对表达量分别为1.00±0.10、1.00±0.13、0.52±0.06,差异有统计学意义(P＜0.05);MMP-2蛋白相对表达量分别为1.00±0.12、0.92±0.08、0.21±0.02,MMP-9蛋白相对表达量分别为1.00±0.13、1.12±0.07、0.17±0.01,表明AFAP1 L2 siRNA转染组MMP-2和MMP-9蛋白表达降低(P＜0.05).侵袭和迁移实验结果显示:空白对照组、无义序列siRNA转染组、AFAP1 L2 siRNA转染组Transwell侵袭实验穿膜细胞数分别为(246.74±10.52)、(236.85±11.74)、(40.14±8.32)个,Transwell迁移实验穿膜细胞数分别为(258.36±11.46)、(230.52±9.87)、(56.43±7.16)个,细胞划痕实验48 h迁移到空白处的细胞数分别为(296.60±27.14)、(113.80±25.43)、(51.20±13.25)个,说明转染AFAP1L2 siRNA组细胞侵袭和迁移能力明显低于其他两组(P＜0.05).结论 沉默胶质瘤细胞中AFAP1L2的表达能显著抑制肿瘤细胞的侵袭和迁移能力.
目的 觀察下調膠質瘤細胞株LN229中微絲相關蛋白1L2 (AFAP1L2)的錶達對細胞侵襲和遷移能力的影響.方法 將LN229細胞分為空白對照組、無義序列小于擾RNA (siRNA)轉染組和AFAP1 L2 siRNA轉染組進行細胞培養,細胞轉染siRNA後,採用Transwell、劃痕實驗檢測細胞侵襲和遷移能力;採用Western blot檢測AFAP1L2、基質金屬蛋白酶(MMP)-2、MMP-9的錶達水平.結果 轉染siRNA後,Western blot檢測結果顯示:空白對照組、無義序列siRNA轉染組、AFAP1L2 siRNA轉染組AFAP1L2蛋白相對錶達量分彆為1.00±0.10、1.00±0.13、0.52±0.06,差異有統計學意義(P＜0.05);MMP-2蛋白相對錶達量分彆為1.00±0.12、0.92±0.08、0.21±0.02,MMP-9蛋白相對錶達量分彆為1.00±0.13、1.12±0.07、0.17±0.01,錶明AFAP1 L2 siRNA轉染組MMP-2和MMP-9蛋白錶達降低(P＜0.05).侵襲和遷移實驗結果顯示:空白對照組、無義序列siRNA轉染組、AFAP1 L2 siRNA轉染組Transwell侵襲實驗穿膜細胞數分彆為(246.74±10.52)、(236.85±11.74)、(40.14±8.32)箇,Transwell遷移實驗穿膜細胞數分彆為(258.36±11.46)、(230.52±9.87)、(56.43±7.16)箇,細胞劃痕實驗48 h遷移到空白處的細胞數分彆為(296.60±27.14)、(113.80±25.43)、(51.20±13.25)箇,說明轉染AFAP1L2 siRNA組細胞侵襲和遷移能力明顯低于其他兩組(P＜0.05).結論 沉默膠質瘤細胞中AFAP1L2的錶達能顯著抑製腫瘤細胞的侵襲和遷移能力.
목적 관찰하조효질류세포주LN229중미사상관단백1L2 (AFAP1L2)적표체대세포침습화천이능력적영향.방법 장LN229세포분위공백대조조、무의서렬소우우RNA (siRNA)전염조화AFAP1 L2 siRNA전염조진행세포배양,세포전염siRNA후,채용Transwell、화흔실험검측세포침습화천이능력;채용Western blot검측AFAP1L2、기질금속단백매(MMP)-2、MMP-9적표체수평.결과 전염siRNA후,Western blot검측결과현시:공백대조조、무의서렬siRNA전염조、AFAP1L2 siRNA전염조AFAP1L2단백상대표체량분별위1.00±0.10、1.00±0.13、0.52±0.06,차이유통계학의의(P＜0.05);MMP-2단백상대표체량분별위1.00±0.12、0.92±0.08、0.21±0.02,MMP-9단백상대표체량분별위1.00±0.13、1.12±0.07、0.17±0.01,표명AFAP1 L2 siRNA전염조MMP-2화MMP-9단백표체강저(P＜0.05).침습화천이실험결과현시:공백대조조、무의서렬siRNA전염조、AFAP1 L2 siRNA전염조Transwell침습실험천막세포수분별위(246.74±10.52)、(236.85±11.74)、(40.14±8.32)개,Transwell천이실험천막세포수분별위(258.36±11.46)、(230.52±9.87)、(56.43±7.16)개,세포화흔실험48 h천이도공백처적세포수분별위(296.60±27.14)、(113.80±25.43)、(51.20±13.25)개,설명전염AFAP1L2 siRNA조세포침습화천이능력명현저우기타량조(P＜0.05).결론 침묵효질류세포중AFAP1L2적표체능현저억제종류세포적침습화천이능력.
Objective To investigate the influence of actin filament associated protein 1-like 2 (AFAP1L2) on invasion and migration of glioma cell LN229.Methods LN229 cells were divided into the control group,nonsense sequence small interfering RNA (siRNA) transfected group and AFAP1L2 siRNA transfected group.The invasion and migration changes of LN229 cells transfected by siRNA were evaluated by transwell assay and scratch assay.After siRNAs were transfected into LN229 cells,the expression of AFAP1L2,matrix metalloproteinase (MMP)-2 and MMP-9 were detected by Western blotting.Results After transfection with siRNA,the results of Western blotting showed that the relative protein expression levels of AFAP1L2,MMP-2 and MMP-9 were 0.52 ±0.06,0.21 ±0.02 and 0.17 ±0.01 in the AFAP1L2 siRNA transfected group,1.00 ±0.10,1.00 ±0.12 and 1.00 ±0.13 in the control group,1.00 ±0.13,0.92 ±0.08 and 1.12 ±0.07 in the nonsense sequence group respectively (P ＜0.05).The results of the transwell assay showed that the invasive and migrated membrane cell number were 40.14 ± 8.32 and 56.43 ±7.16 in the AFAP1L2 siRNA transfected group,246.74 ± 10.52 and 258.36 ± 11.46 in the control group,236.85 ± 11.74 and 230.52 ± 9.87 in the nonsense sequence group respectively (P ＜ 0.05).Results of wound healing assay in the control group,nonsense sequence group,AFAP1L2 siRNA transfected group were 296.60 ± 27.14,113.80 ± 25.43 and 51.20 ± 13.25 respectively (P ＜ 0.05).The transwell assay and scratch assay results showed that the ability of invasion and migration were significantly reduced in AFAP1 L2 siRNA transfected group (P ＜ 0.05).Conclusion Using RNAi technology,siRNA targeting AFAP1L2 efficiently knocks down the expression of AFAP1L2 in human glioma cells,results in suppression of tumor cell invasion and migration.AFAP1L2 is a potential therapeutic target for malignant gliomas.
