中国医师杂志
中國醫師雜誌
중국의사잡지
JOURNAL OF CHINESE PHYSICIAN
2014年
11期
1508-1512
,共5页
基因沉默%脑肿瘤/病理学%神经胶质瘤/病理学%ATP结合匣式转运子/遗传学%肿瘤细胞,培养的%转染%细胞增殖%细胞运动
基因沉默%腦腫瘤/病理學%神經膠質瘤/病理學%ATP結閤匣式轉運子/遺傳學%腫瘤細胞,培養的%轉染%細胞增殖%細胞運動
기인침묵%뇌종류/병이학%신경효질류/병이학%ATP결합갑식전운자/유전학%종류세포,배양적%전염%세포증식%세포운동
Gene silencing%Brain neoplasms/pathology%Glioma/pathology%ATP-binding cassette transporters/genetics%Tumor cells,cultured%Transfection%Cell proliferation%Cell movement
目的 通过电转法沉默ATP结合盒转运子E1(ABCE1)基因在人脑胶质瘤U87MG细胞中的表达,探讨ABCE1基因对细胞的生长、细胞周期及肿瘤迁移等方面的作用.方法 设计及合成ABCE1的siRNA序列,电转法转染U87MG细胞.通过RT-PCR、Western blot检测ABCE1在电转后的mRNA和蛋白的表达情况,用流式细胞仪检测细胞周期及凋亡.通过CCK-8增殖实验、划痕愈合实验、细胞侵袭实验评价沉默ABCE1基因对人脑胶质瘤U87MG细胞增殖、迁移、以及侵袭能力的影响.结果 ABCE1 mRNA和蛋白表达,实验组显著低于对照组和空白组,差异有统计学意义(P＜0.05).实验组细胞的生长速度明显减慢,细胞周期被阻滞在G0/G1期,S期细胞数减少,与对照组和空白组比较差异有统计学意义(P＜0.05);细胞凋亡率明显高于对照组和空白组,差异有统计学意义(P＜0.01).与对照组和空白组相比实验组U87MG细胞的增殖受到明显抑制(P＜0.05).实验组迁移能力显著下降,差异有统计学意义(P＜0.05);实验组U87MG细胞的侵袭能力显著下降,差异有统计学意义(P＜0.01).结论 ABCE1与胶质瘤细胞的增殖、侵袭和转移能力密切相关,应用电转法沉默ABCE1基因可以明显降低U87MG细胞体外增殖、侵袭及转移能力.
目的 通過電轉法沉默ATP結閤盒轉運子E1(ABCE1)基因在人腦膠質瘤U87MG細胞中的錶達,探討ABCE1基因對細胞的生長、細胞週期及腫瘤遷移等方麵的作用.方法 設計及閤成ABCE1的siRNA序列,電轉法轉染U87MG細胞.通過RT-PCR、Western blot檢測ABCE1在電轉後的mRNA和蛋白的錶達情況,用流式細胞儀檢測細胞週期及凋亡.通過CCK-8增殖實驗、劃痕愈閤實驗、細胞侵襲實驗評價沉默ABCE1基因對人腦膠質瘤U87MG細胞增殖、遷移、以及侵襲能力的影響.結果 ABCE1 mRNA和蛋白錶達,實驗組顯著低于對照組和空白組,差異有統計學意義(P＜0.05).實驗組細胞的生長速度明顯減慢,細胞週期被阻滯在G0/G1期,S期細胞數減少,與對照組和空白組比較差異有統計學意義(P＜0.05);細胞凋亡率明顯高于對照組和空白組,差異有統計學意義(P＜0.01).與對照組和空白組相比實驗組U87MG細胞的增殖受到明顯抑製(P＜0.05).實驗組遷移能力顯著下降,差異有統計學意義(P＜0.05);實驗組U87MG細胞的侵襲能力顯著下降,差異有統計學意義(P＜0.01).結論 ABCE1與膠質瘤細胞的增殖、侵襲和轉移能力密切相關,應用電轉法沉默ABCE1基因可以明顯降低U87MG細胞體外增殖、侵襲及轉移能力.
목적 통과전전법침묵ATP결합합전운자E1(ABCE1)기인재인뇌효질류U87MG세포중적표체,탐토ABCE1기인대세포적생장、세포주기급종류천이등방면적작용.방법 설계급합성ABCE1적siRNA서렬,전전법전염U87MG세포.통과RT-PCR、Western blot검측ABCE1재전전후적mRNA화단백적표체정황,용류식세포의검측세포주기급조망.통과CCK-8증식실험、화흔유합실험、세포침습실험평개침묵ABCE1기인대인뇌효질류U87MG세포증식、천이、이급침습능력적영향.결과 ABCE1 mRNA화단백표체,실험조현저저우대조조화공백조,차이유통계학의의(P＜0.05).실험조세포적생장속도명현감만,세포주기피조체재G0/G1기,S기세포수감소,여대조조화공백조비교차이유통계학의의(P＜0.05);세포조망솔명현고우대조조화공백조,차이유통계학의의(P＜0.01).여대조조화공백조상비실험조U87MG세포적증식수도명현억제(P＜0.05).실험조천이능력현저하강,차이유통계학의의(P＜0.05);실험조U87MG세포적침습능력현저하강,차이유통계학의의(P＜0.01).결론 ABCE1여효질류세포적증식、침습화전이능력밀절상관,응용전전법침묵ABCE1기인가이명현강저U87MG세포체외증식、침습급전이능력.
Objective To investigate the effect of ATP-binding cassette protein E 1 (ABCE1) gene silencing by electroporation on the survival,cell cycle and invasion of human glioma cells line U87MG.Methods The siRNA against ABCE1 was constructed and transfected into U87MG cells by electroporation.The expression of ABCE1 was detected by real time-polymerase chain reaction (RT-PCR) and Western blot.Flow cytometry was used to detect the cell cycle distribution and apoptosis.The effects of ABCE1 gene silencing by electroporation on proliferation,migration and invasion of U87MG cell line were evaluated by cell counting kit-8 (CCK-8) assay,wound closure assay,chemotactic migration,and cell invasive experiments,respectively.Results Compared to the control and blank groups,the mRNA and protein levels were significantly decreased in the experimental group when ABCE1 gene silencing by electroporation.The cell cycle was arrested at G0/G1 phase,and cell number in S phase was decreased in U87MG cell line (P ＜ 0.05).The cell growth inhibition ratio in the experimental group was significantly higher than that in the control and blank groups (P ＜0.01).Compared to the control and blank groups,the experimental group U87MG cell proliferation was inhibited significantly (P ＜ 0.05).Scratch healing experiments showed the experimental group migration ability was decreased significantly (P ＜ 0.05).Transwell chamber method showed the experimental group U87MG cell invasion ability was decreased significantly (P ＜ 0.05).Conclusions ABCE1 is involved in the progression of human glioma cells,and inhibiting the expression of ABCE1 by electroporation can decrease migration,invasion,and proliferation ability of tumor cells in vitro.
