中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
Chinese Journal of Experimental Surgery
2015年
9期
2077-2080
,共4页
张芸%朱一硕%梅鹏金%陈晨%范月超
張蕓%硃一碩%梅鵬金%陳晨%範月超
장예%주일석%매붕금%진신%범월초
胶质瘤%三磷酸腺苷结合盒转运子E1%核糖核酸酶L
膠質瘤%三燐痠腺苷結閤盒轉運子E1%覈糖覈痠酶L
효질류%삼린산선감결합합전운자E1%핵당핵산매L
Glioma%Adenosine triphosphate binding cassette E1%RNase L
目的 观察三磷酸腺苷(ATP)结合盒转运子E1基因(ABCEl)在胶质瘤及正常脑组织中的表达及其对人脑胶质瘤细胞U251的增殖和凋亡的影响.方法 选取45例胶质瘤和10例正常脑组织(脑外伤患者内减压切除的脑组织)作为研究对象,应用免疫组织化学技术检测ABCE1的表达水平.另选取7例胶质瘤组织和4例正常脑组织作为研究对象,应用Western blot技术检测胶质瘤组织和正常脑组织之间ABCE1表达量的差异.此外,应用小干扰RNA (siRNA)技术转染脑胶质瘤U251细胞,Western blot检测ABCE1 siRNA对ABCE1蛋白的干扰效果和其靶蛋白核糖核酸酶L(RNase L)的表达,同时应用细胞计数试剂盒(CCK-8)细胞增殖实验和凋亡实验检测ABCE1 siRNA对U251细胞增殖和凋亡的影响.结果 免疫组织化学染色结果显示ABCE1在正常组、Ⅰ～Ⅱ级组和Ⅲ～Ⅳ级中表达差异有统计学意义(P＜0.05),且ABCE1的表达量与肿瘤的恶性程度呈正相关(r=0.626,P＜0.05).Western blot检测表明ABCE1随着肿瘤恶性程度的增高表达量也随之增高,3组之间差异均有统计学意义(P＜0.05).ABCE1的siRNA转染U251细胞后明显抑制细胞增殖,在48、72、96 h吸光度值分别降低了0.394、0.542、0.968;同时,ABCE1 siRNA组凋亡率为(12.730±0.580)％,与阴性对照组的(4.800±0.118)％和空白组的(3.830±0.086)％比较,其凋亡率均明显增高(P＜0.05);当ABCE1下调后RNase L的表达量增加,干扰组与空白、阴性对照组比较差异均有统计学意义(P＜0.05).结论 ABCE1在胶质瘤和正常脑组织中均表达,其在胶质瘤中的表达水平与其恶性程度呈正相关.ABCE1在胶质瘤细胞U251的增殖和凋亡方面扮演重要角色,同时通过特异性抑制2-5A/RNase L通路,干扰胶质瘤细胞的生物特点.
目的 觀察三燐痠腺苷(ATP)結閤盒轉運子E1基因(ABCEl)在膠質瘤及正常腦組織中的錶達及其對人腦膠質瘤細胞U251的增殖和凋亡的影響.方法 選取45例膠質瘤和10例正常腦組織(腦外傷患者內減壓切除的腦組織)作為研究對象,應用免疫組織化學技術檢測ABCE1的錶達水平.另選取7例膠質瘤組織和4例正常腦組織作為研究對象,應用Western blot技術檢測膠質瘤組織和正常腦組織之間ABCE1錶達量的差異.此外,應用小榦擾RNA (siRNA)技術轉染腦膠質瘤U251細胞,Western blot檢測ABCE1 siRNA對ABCE1蛋白的榦擾效果和其靶蛋白覈糖覈痠酶L(RNase L)的錶達,同時應用細胞計數試劑盒(CCK-8)細胞增殖實驗和凋亡實驗檢測ABCE1 siRNA對U251細胞增殖和凋亡的影響.結果 免疫組織化學染色結果顯示ABCE1在正常組、Ⅰ～Ⅱ級組和Ⅲ～Ⅳ級中錶達差異有統計學意義(P＜0.05),且ABCE1的錶達量與腫瘤的噁性程度呈正相關(r=0.626,P＜0.05).Western blot檢測錶明ABCE1隨著腫瘤噁性程度的增高錶達量也隨之增高,3組之間差異均有統計學意義(P＜0.05).ABCE1的siRNA轉染U251細胞後明顯抑製細胞增殖,在48、72、96 h吸光度值分彆降低瞭0.394、0.542、0.968;同時,ABCE1 siRNA組凋亡率為(12.730±0.580)％,與陰性對照組的(4.800±0.118)％和空白組的(3.830±0.086)％比較,其凋亡率均明顯增高(P＜0.05);噹ABCE1下調後RNase L的錶達量增加,榦擾組與空白、陰性對照組比較差異均有統計學意義(P＜0.05).結論 ABCE1在膠質瘤和正常腦組織中均錶達,其在膠質瘤中的錶達水平與其噁性程度呈正相關.ABCE1在膠質瘤細胞U251的增殖和凋亡方麵扮縯重要角色,同時通過特異性抑製2-5A/RNase L通路,榦擾膠質瘤細胞的生物特點.
목적 관찰삼린산선감(ATP)결합합전운자E1기인(ABCEl)재효질류급정상뇌조직중적표체급기대인뇌효질류세포U251적증식화조망적영향.방법 선취45례효질류화10례정상뇌조직(뇌외상환자내감압절제적뇌조직)작위연구대상,응용면역조직화학기술검측ABCE1적표체수평.령선취7례효질류조직화4례정상뇌조직작위연구대상,응용Western blot기술검측효질류조직화정상뇌조직지간ABCE1표체량적차이.차외,응용소간우RNA (siRNA)기술전염뇌효질류U251세포,Western blot검측ABCE1 siRNA대ABCE1단백적간우효과화기파단백핵당핵산매L(RNase L)적표체,동시응용세포계수시제합(CCK-8)세포증식실험화조망실험검측ABCE1 siRNA대U251세포증식화조망적영향.결과 면역조직화학염색결과현시ABCE1재정상조、Ⅰ～Ⅱ급조화Ⅲ～Ⅳ급중표체차이유통계학의의(P＜0.05),차ABCE1적표체량여종류적악성정도정정상관(r=0.626,P＜0.05).Western blot검측표명ABCE1수착종류악성정도적증고표체량야수지증고,3조지간차이균유통계학의의(P＜0.05).ABCE1적siRNA전염U251세포후명현억제세포증식,재48、72、96 h흡광도치분별강저료0.394、0.542、0.968;동시,ABCE1 siRNA조조망솔위(12.730±0.580)％,여음성대조조적(4.800±0.118)％화공백조적(3.830±0.086)％비교,기조망솔균명현증고(P＜0.05);당ABCE1하조후RNase L적표체량증가,간우조여공백、음성대조조비교차이균유통계학의의(P＜0.05).결론 ABCE1재효질류화정상뇌조직중균표체,기재효질류중적표체수평여기악성정도정정상관.ABCE1재효질류세포U251적증식화조망방면분연중요각색,동시통과특이성억제2-5A/RNase L통로,간우효질류세포적생물특점.
Objective To investigate the expression of adenosine triphosphate (ATP) binding cassette E1 (ABCE1) in glioma and its significance in proliferation and apoptosis of the glioma U251 cell line.Methods Expression of ABCE1 in 45 cases of glioma tissues and 10 cases normal brain (NB) tissues was analyzed by immunohis-tochemistry.Meanwhile,Expression of ABCE1 in 11 cases of tissues was analyzed by western blot.Furthermore,small interfering RNA (siRNA) targeting ABCE1 was constructed and transfected into U251 human glioma cells to downregulate ABCE1 expression.The effect of ABCE1 knockdown on cell proliferation,apoptosis and gene expression was then assessed using CCK-8 assay,flow cytometry and western blot analysis,respectively.Results Our results showed that ABCE1 expression gradually increased in normal brain,grades Ⅰ-Ⅱ and grades Ⅲ-Ⅳ tumor (P ＜ 0.05).Mean-while,a positive relationship between the level of ABCE1 and the malignancy of glioma was determined (r =0.626,P ＜ 0.05).As compared with negative control group,cell proliferation ability was decreased with the absorbance values being respectively decreased by 0.394,0.542,0.968 at 48,72,96 h;The apoptosis rate in ABCE1 siRNA group (12.730 ± 0.580) ％ was significantly increased as compared with blank group (3.830 ±0.086)％and negative control group (4.800 ±0.118)％ (P ＜0.05).Meanwhile,the expression of ABCE1 in U251 was blocked (P ＜ 0.05),while the expression of RNase L increased significantly (P ＜ 0.05).Conclusion The presence of ABCE1 was widely detected in both normal human brain and glioma tissues.Its level was positively associated with its malignancy.These findings showed that ABCE1 had an important role in proliferation and apoptosis in U251 human glioma cells and that ABCE1 may inhibit intracellular RNase L activity,which inhibits the 2-5A/RNase L pathway,interfering with the biological characteristics of glioma cells.
