中华神经医学杂志
中華神經醫學雜誌
중화신경의학잡지
CHINESE JOURNAL OF NEUROMEDICINE
2015年
6期
553-557
,共5页
陈斯泽%张威%钟德泉%陈光忠%陈雪梅
陳斯澤%張威%鐘德泉%陳光忠%陳雪梅
진사택%장위%종덕천%진광충%진설매
蛋白半胱氨酸蛋白酶-3%Gpc1%神经胶质瘤%增殖
蛋白半胱氨痠蛋白酶-3%Gpc1%神經膠質瘤%增殖
단백반광안산단백매-3%Gpc1%신경효질류%증식
Caspase-3%Glypicante1%Glioma%Proliferation
目的 研究凋亡相关蛋白半胱氨酸蛋白酶(Caspase)-3及Glypicante1 (Gpc1)的表达对神经胶质瘤细胞增殖能力的影响及其可能机制. 方法 取广东省人民医院神经外科自2011年1月至2011年12月手术切除的52例胶质瘤标本和同期因脑外伤行颅内减压术的13例患者的正常脑组织标本,免疫组化染色检测标本中Caspase-3和Gpc1的表达;体外常规培养人神经胶质瘤A172细胞株,构建peDNA3.1/Gpc1高表达质粒,Luciferase双荧光素酶报告基因试剂盒检测0.5、1、2、4 μg peDNA3.1/Gpc1高表达质粒转染A172细胞后Hedgehog (Hh)信号通路活性的变化;用siRNA-Gpc1干扰质粒转染A172细胞沉默Gpc1的表达,再使用8μg/mL抗生素杀稻瘟菌素S处理细胞24h(实验组),以加入等量溶剂的正常A172细胞作为对照组,免疫组化染色检测2组细胞CyclinD1的表达,噻唑蓝(MTT)检测转染后24、48、72和96h2组细胞的增殖活力. 结果 神经胶质瘤组织Caspase-3、Gpc1表达阳性率均高于正常脑组织,差异有统计学意义(P＜0.05),神经胶质瘤标本中Caspase-3与Gpc1的表达呈正相关关系(r=0.486,P=0.000);与对照组比较,0.5、1、2、4μg peDNA3.1/Gpc1高表达质粒组细胞荧光强度增加,差异有统计学意义(P＜0.05);免疫组化染色显示沉默Gpc1的表达后A172细胞Hh通路中靶基因CyclinD1的表达低于正常A172细胞.MTT检测显示沉默Gpc1的表达后48、72、96h A172细胞株的增殖活力均低于对照组,差异有统计学意义(P＜0.05). 结论 Caspase-3与Gpc1蛋白的表达异常在神经胶质瘤细胞增殖中起重要作用,可能与Hh信号通路的介导有关.
目的 研究凋亡相關蛋白半胱氨痠蛋白酶(Caspase)-3及Glypicante1 (Gpc1)的錶達對神經膠質瘤細胞增殖能力的影響及其可能機製. 方法 取廣東省人民醫院神經外科自2011年1月至2011年12月手術切除的52例膠質瘤標本和同期因腦外傷行顱內減壓術的13例患者的正常腦組織標本,免疫組化染色檢測標本中Caspase-3和Gpc1的錶達;體外常規培養人神經膠質瘤A172細胞株,構建peDNA3.1/Gpc1高錶達質粒,Luciferase雙熒光素酶報告基因試劑盒檢測0.5、1、2、4 μg peDNA3.1/Gpc1高錶達質粒轉染A172細胞後Hedgehog (Hh)信號通路活性的變化;用siRNA-Gpc1榦擾質粒轉染A172細胞沉默Gpc1的錶達,再使用8μg/mL抗生素殺稻瘟菌素S處理細胞24h(實驗組),以加入等量溶劑的正常A172細胞作為對照組,免疫組化染色檢測2組細胞CyclinD1的錶達,噻唑藍(MTT)檢測轉染後24、48、72和96h2組細胞的增殖活力. 結果 神經膠質瘤組織Caspase-3、Gpc1錶達暘性率均高于正常腦組織,差異有統計學意義(P＜0.05),神經膠質瘤標本中Caspase-3與Gpc1的錶達呈正相關關繫(r=0.486,P=0.000);與對照組比較,0.5、1、2、4μg peDNA3.1/Gpc1高錶達質粒組細胞熒光彊度增加,差異有統計學意義(P＜0.05);免疫組化染色顯示沉默Gpc1的錶達後A172細胞Hh通路中靶基因CyclinD1的錶達低于正常A172細胞.MTT檢測顯示沉默Gpc1的錶達後48、72、96h A172細胞株的增殖活力均低于對照組,差異有統計學意義(P＜0.05). 結論 Caspase-3與Gpc1蛋白的錶達異常在神經膠質瘤細胞增殖中起重要作用,可能與Hh信號通路的介導有關.
목적 연구조망상관단백반광안산단백매(Caspase)-3급Glypicante1 (Gpc1)적표체대신경효질류세포증식능력적영향급기가능궤제. 방법 취광동성인민의원신경외과자2011년1월지2011년12월수술절제적52례효질류표본화동기인뇌외상행로내감압술적13례환자적정상뇌조직표본,면역조화염색검측표본중Caspase-3화Gpc1적표체;체외상규배양인신경효질류A172세포주,구건peDNA3.1/Gpc1고표체질립,Luciferase쌍형광소매보고기인시제합검측0.5、1、2、4 μg peDNA3.1/Gpc1고표체질립전염A172세포후Hedgehog (Hh)신호통로활성적변화;용siRNA-Gpc1간우질립전염A172세포침묵Gpc1적표체,재사용8μg/mL항생소살도온균소S처리세포24h(실험조),이가입등량용제적정상A172세포작위대조조,면역조화염색검측2조세포CyclinD1적표체,새서람(MTT)검측전염후24、48、72화96h2조세포적증식활력. 결과 신경효질류조직Caspase-3、Gpc1표체양성솔균고우정상뇌조직,차이유통계학의의(P＜0.05),신경효질류표본중Caspase-3여Gpc1적표체정정상관관계(r=0.486,P=0.000);여대조조비교,0.5、1、2、4μg peDNA3.1/Gpc1고표체질립조세포형광강도증가,차이유통계학의의(P＜0.05);면역조화염색현시침묵Gpc1적표체후A172세포Hh통로중파기인CyclinD1적표체저우정상A172세포.MTT검측현시침묵Gpc1적표체후48、72、96h A172세포주적증식활력균저우대조조,차이유통계학의의(P＜0.05). 결론 Caspase-3여Gpc1단백적표체이상재신경효질류세포증식중기중요작용,가능여Hh신호통로적개도유관.
Objective To explore the effects of Caspase-3 and Glypicante1 (Gpc1) altered expressions on the proliferation of glioma cells and its mechanism.Methods Fifty-two glioma specimens and 13 normal cerebral tissues were collected in our hospital from October 2011 to October 2014;the protein expressions of Caspase-3 and Gpc1 in these tissues were detected with immunohistochemistry.The peDNA3.1/Gpc 1 high expression plasmids were constructed;human glioma A172 cell lines were routinely cultured in vitro;the signal path changes of Hedgehog (H-h) in the A172 cells after being transfected with 0.5,1,2 and 4 μg peDNA3.1/Gpc1 high expression plasmids were detected by Luciferase luciferase reporter gene kit.Gpc 1-1694 interference plasmids were transfected into the A172 cells to silence the Gpc1 expression and these A172 cells were given 8 iμg/mL Blasticidin (experimental group) for 24 h;normal A172 cells were given the same volume of solvent (control group);altered expressions of G-pc 1 were detected by immumohistochemical staining and proliferation activity of the cells were observed by MTT assay 24,48,72 and 96 h after transfection.Results The expressions of Caspase-3 and Gpc1 in the glioma specimens were significantly higher than those in the normal brain tissues (P＜0.05);in the glioma specimens,positive correlation between Caspase-3 and Gpc1 was noted (r=0.486,P=0.000).As compared with the control group,increased fluorescence intensity was observed in the 0.5,1,2 and 4 μg peDNA3.1/Gpc1 high expression plasmids groups (P＜0.05);immumohistochemical staining showed that CyclinD1 expression in the Hh pathway of in the peDNA 3.1/Gpc1 high expression plasmids groups was significantly lower than that in the control group;MTT assay showed that the proliferation activity of the A172 cells in the peDNA3.1/Gpc1 high expression plasmids groups at 24,48,72 and 96 h after transfection was significantly lower than that in the control group (P＜0.05).Conclusion Caspase-3 and Gpc1 play important roles in regulating the proliferation of glioma cells,which might be related to Hedgehog signaling pathway.
